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Volume I
British Pharmacopoeia 2016
Volume I
The British Pharmacopoeia Commission has caused this British
Pharmacopoeia 2016 to be prepared under regulation 317(1) of the Human
Medicines Regulations 2012 and, in accordance with regulation 317(4), the
Ministers have arranged for it to be published. It has been notified in draft
to the European Commission in accordance with Directive 98/34/EEC.
The monographs of the Eighth Edition of the European Pharmacopoeia
(2013), as amended by Supplements 8.1 to 8.5, published by the Council
of Europe are reproduced either in this edition of the British
Pharmacopoeia or in the associated edition of the British Pharmacopoeia
(Veterinary).
See General Notices
The British Pharmacopoeia (BP), after 150 years of publication, continues to help ensure the quality of
medicinal substances globally. One of its key attributes has been to take advantage of novel science and
this edition of the BP is no different.
Authentication of botanical constituents, to ensure the safety and quality of Traditional Herbal Medicines,
provides many challenges. With advances in molecular genetics, however, reliable methods of identifying
herbs are now available. This has enabled the BP and the National Institute for Biological Standards and
Control (NIBSC) to work collaboratively and successfully on a pilot project to determine the
deoxyribonucleic acid (DNA) profile of Ocimum tenuiflorum.
The BP 2016 introduces the species specific sequence of the selected barcode region in a new BP
Appendix as an identification method for Ocimum tenuiflorum. General guidance on how to conduct
DNA-based identification methods for herbal drugs is also included in this new Appendix. In addition to this
innovation, the BP 2016 publishes new and revised monographs for substances and products used in a
wide range of medicines. The collegial partnerships between the staff, the appointed experts of the British
Pharmacopoeia Commission and international partners have enabled these important quality standards to
be made publicly available.
The significant progress made jointly by the BP and NIBSC facilitates the reliable authentication of herbal
drugs and will enhance the protection of public health, the primary objective of the Medicines and
Healthcare products Regulatory Agency.
Contents of Volume I
FOREWORD
NOTICES
PREFACE
BRITISH PHARMACOPOEIA COMMISSION
EXPERT ADVISORY GROUPS, PANELS OF EXPERTS AND
WORKING PARTIES
CODE OF PRACTICE
MEMBERSHIP
BP Commission, Expert Advisory Groups, Panels of Experts, Working
Parties
STAFF
British Pharmacopoeia, BP Laboratory, Publisher
INTRODUCTION
Additions, Omissions, Technical Changes, Changes in Tide
GENERAL NOTICES
MONOGRAPHS
Medicinal and Pharmaceutical Substances (A - 1)
Contents of Volume II
NOTICES
GENERAL NOTICES
MONOGRAPHS
Medicinal and Pharmaceutical Substances Q - Z)
Contents of Volume m
NOTICES
GENERAL NOTICES
MONOGRAPHS
Formulated Preparations: General Monographs
Formulated Preparations: Specific Monographs
I-vii
Contents of Volume IV
NOTICES
GENERAL NOTICES
MONOGRAPHS
Herbal Drugs, Herbal Drug Preparations and Herbal Medicinal Products
Materials for use in the Manufacture of Homoeopathic Preparations
Blood-related Products
Immunological Products
Radiopharmaceutical Preparations
Surgical Materials
Contents of Volume V
NOTICES
GENERAL NOTICES
INFRARED REFERENCE SPECTRA
APPENDICES
SUPPLEMENTARY CHAPTERS
INDEX
Notices
I-ix
Preface
(4) to approve new and revised text for inclusion in new editions of the
British Pharmacopoeia and British Pharmacopoeia (Veterinary);
(5) to approve new and revised names for inclusion in new editions of
British Approved Names and its annual supplements.
I-xi
In addition to the duties listed above, the Chair of the British
Pharmacopoeia Commission has the following additional duties:
(1) To chair all scheduled and unscheduled meetings;
(2) To carry out members appraisals in accordance with Department of
Health policies and timelines;
(3) To participate in the process to appoint/re-appoint members of the
British Pharmacopoeia Commission.
Expert Advisory Groups, Panels
o f Experts and Working Parties
I-xiii
Code of Practice
The list below includes those members who served during the period 2014
to 2015.
Chair Professor Kevin M G Taylor BPharm PhD FRPharmS
Professor of Clinical Pharmaceutics, UCL School of Pharmacy
Vice-Chair Professor Alastair Davidson BSc PhD FRPharmS
Visiting Professor of Pharmaceutical Sciences, University of Strathclyde
Professor Donald Cairns BSc PhD MRPharmS CSci CChem FRSC
Head: School of Pharmacy and Life Sciences, Robert Gordon University,
Aberdeen
Mr Barry Capon CBE MA DL {Lay representative)
Former Non-executive Director, Norfolk and Suffolk NHS Foundation Trust
Dr Graham D Cook BPharm PhD MRPharmS
Senior Director, Process Knowledge!Quality by Design, Pfizer
Mr Andrew Coulson BVetMed MSc MRCVS
Member of the Royal College of Veterinary Surgeons; former Superintending
Inspector, Science & Research Group, The Home Office
Mr Christopher Goddard BSc DIS CSci EurChem CChem FRSC
Quality Control Technical Manager, Recipharm Limited
Dr Keith Helliwell BPharm PhD
Senior Technical Adviser, William Ransom & Son PLC
Dr Rodney L Horder BPharm PhD MRPharmS
Former Divisional Vice President, European Quality and Regulatory Strategy,
Abbott
Dr Gerard Lee BPharm PhD FRPharmS MRSC CChem
Former Group Manager, British Pharmacopoeia and Laboratory Services
(MHRA); former Secretary & Scientific Director of the British Pharmacopoeia
Commission
Dr Brian R Matthews BPharm PhD FRPharmS FTOPRA MRI
Consultant on pharmaceutical and medical device regulatory affairs; former Senior
Director, EC Registration, Alcon Laboratories
I-xv
Dr Lincoln Tsang BPharm LLB PhD FRSC FIBiol FRSA FRPharmS
Solicitor
Life Sciences Lawyer; Partner, Arnold & Porter LLP
Mrs Josephine Turnbull T-T R {Lay representative)
Former Chair of Tees, Esk and Wear Valley NHS Foundation Trust
Dr Paul Varley BSc PhD
Vice President of Biopharmaceutical Development, Medimmune Limited
Professor Elizabeth Williamson BPharm PhD MRPharmS
Former Professor of Pharmacy, University of Reading
Secretary and Scientific Dr Samantha Atkinson BSc MSc PhD MRSC
Director Visiting Fellow, University of Reading
I-xvi
Membership of Expert Advisory
Groups, Panels of Experts and
Working Parties
I-xvii
ULM: Unlicensed M G Lee (Chair), V Fenton-May (Vice-Chair), G Bennett, S Branch,
Medicines D Caulfield, A Charvill, W Goddard, N Hussain, S Jones, M A Oldcome,
N J Precious, J Rothwell, M Santdllo, J Smith, A Sully, P Weir
PANELS OF EXPERTS
BLP: Blood K Chidwick, A R Hubbard, J More, P Varley
Products
CX: Excipients B R Matthews (Chair), C Mroz (Vice-Chair), E Anno, C Cable, R
Cawthome, W Cook, D Deutsch, N Hussain, M I Robertson, K Slevin
IGC: Inorganic and C T Goddard (Chair), M Almond, S Atherton, S Boland, A C Cartwright,
General Chemicals D Caulfield, P Henrys, G Lay, D Malpas, C Mroz, D Riches
MIC: Microbiology V Fenton-May (Chair), S Denyer, D P Hargreaves, B R Matthews,
P Newby
RAD: Radioactive J Ballinger, J Brain, D Graham, S R Hesslewood, G Inwards, P Maltby, R
Materials D Pickett, R Smith, S Waters
VET: Veterinary E Williamson (Chair), A Coulson (Vice-Chair), A Cairns, S Cockbill,
Medicines D Evans, E Flahive, P Lees, B Ward
VIP: Veterinary A M Brady, R Banks, K Redhead, J Salt, P W Wells, R Woodland
Immunological
Products
WORKING PARTIES
AQbD: Analytical G Cook (Chair), S Brown, S Ellison, M Hanna-Brown, S Jones,
Quality by Design D Makohon, P Nethercote, E Razzano
(Corresponding member K Barnett)
DNA: Identification K Helliwell (Chair), J Hawkins, E Mee, A Slater, E Williamson
Techniques
MCS: Microscopy E Williamson (Chair), R Arroo, R Reck, K Helliwell, K MacLellan Gibson
I-xviii
Current British Pharmacopoeia
Staff
ISO 9001
FS 27268
Current British Pharmacopoeia
Laboratory Staff
ISO 9001
FS 27613
I-xx
Current Staff of the Publisher of
the British Pharmacopoeia
ISO 9001
FS 22428
I-xxi
2016 Introduction I-xxiii
Introduction
Effective Date The effective date for British Pharmacopoeia monographs in this edition is
1 January 2016.
Additions A list of monographs included for the first time in the British
Pharmacopoeia 2016 is given at the end of this introduction. It includes 37
new monographs of national origin and 25 new monographs reproduced
from the 8th Edition of the European Pharmacopoeia as amended by
Supplements 8.1 to 8.5.
Traditional H erbal M edicines; Homoeopathic Preparations
Work is continuing on the development of monographs for herbs used in
traditional herbal medicines and homoeopathic medicines. The Latin
scientific names cited in BP monographs for herbal drugs are consistent
with the advice provided by the Medicinal Plant Names Services at the
Royal Botanic Gardens, Kew. As stated in previous editions, the
requirements for the quality of the material are provided in the monograph
to set the standards for Traditional Herbal Medicines in the UK and to
assist the UK Traditional Herbal Medicines Registration Scheme. The
British Pharmacopoeia Commission, however, has not assessed the safety
and efficacy of the materials in traditional use.
2016 Introduction I-xxv
Omissions Thirty five monographs have been omitted from the British Pharmacopoeia
2016. The list of omissions is appended at the end of this Introduction.
In line with recommendations from the Commission on Human Medicines
and the British Pharmacopoeia Commission, reference to chloroform as an
ingredient in licensed medicines has been removed from all affected
monographs in this publication. This has been through either removing the
monograph from the publication or deleting the formula and/or method of
preparation.
Infrared Reference As with the previous edition, the reference spectra are placed in alphabetical
Spectra order within this edition. Two new spectra have been added to the
collection.
Appendices Two new Appendices to harmonise with the European Pharmacopoeia were
first published in the British Pharmacopoeia 2015 electronic updates. These
have been consolidated in the new edition as follows.
Appendix VUIX: Methyl, Ethyl and Isopropyl Toluenesulfonate in Active
Substances (Ph. Eur. method 2.5.40)
Appendix XV K: Carrier Proteins for the Production of Conjugated
Polysaccharide Vaccines for Human Use (Ph. Eur. method 5.2.11)
A new BP method, Appendix XI V: Deoxyribonucleic Acid Based
Identification Techniques for Herbal Drugs, is introduced in this edition
and is accompanied by a worked example for Holy Basil.
Pharmacopoeial It should be noted that any article intended for medicinal use which is
Requirements described by a name at the head of a monograph in the current edition of
the Pharmacopoeia must comply with that monograph *whether or not it is
referred to as BP.
I-xxviii Introduction 2016
Code of Practice Members of the British Pharmacopoeia Commission and its supporting
Expert Advisory Groups, Panels of Experts and Working Parties are
required to comply with a Code of Practice on Declaration of Interests in •
the pharmaceutical industry. Details of the Code are published on the
website (www.pharmacopoeia.com).
Subscribers to the BP online will find that these resources will also be
linked with relevant texts and directly accessible from the BP online
content.
Access to previous editions of the BP will be available as a BP archive
product for purchase by new and existing BP online subscribers. The
content of the archive will start from the BP 2014 onwards and will grow
year-on-year as superseded editions are added to the archive.
Improvements and new features have also been made to the BPCRS
catalogue and ordering facility. Users will be able to receive notifications on
the status of out-of-stock items and view order histories. BPCRS products
will also be linked with relevant BP monographs and subscribers to the BP
online will be able to purchase these directly from the BP online content.
BPCRS customers will continue to be able to make purchases through
invoice or credit card orders.
An email subscription feature will allow users to keep abreast with BP news
and BPCRS updates.
In line with a policy of continuous improvement, users of the newly
consolidated www.pharmacopoeia.com website are invited to provide the
Secretariat with feedback on their experience.
European Pharm acopoeia Websites
httpss://extranet.edqm.eu/publications/recherches_sw.shtml For those texts
reproduced from the European Pharmacopoeia, the EDQM website
provides access to a database (the Knowledge database) containing
information of various sorts related to monographs and intended to
facilitate their proper use. Information is provided on chromatographic
columns used in monograph development, suppliers of reagents and
equipment that may be difficult to find for some users, the status of
monographs (in development, adopted, published, under revision), revisions
of the monographs on a historical basis, beginning from the 5th Edition of
the European Pharmacopoeia as well as other useful information.
https://pharmeuropa.edqm.eu/home The European Pharmacopoeia Forum,
Pharmeuropa, is published quarterly as an aid for the elaboration of
monographs and as a vehicle for information on pharmacopoeial and related
matters. Pharmeuropa is available as a free on-line publication.
Forward Look Electronic Updates The British Pharmacopoeia 2016 online updates will
be published on tile website, www.pharmacopoeia.comj to enable users to
keep up to date with monographs published in tile European
Pharmacopoeia. These updates will be integrated annually with the
publication of the main edition of the British Pharmacopoeia.
Additions The following monographs of the British Pharmacopoeia 2016 were not
included in the British Pharmacopoeia 2015.
Medicinal and Pharm aceutical Substances
Eplerenone*
Glucosamine Sulfate Potassium Chloride*
Imatinib Mesilate*
High-molecular-mass Macrogols*
Macrogol Isotridecyl Ether*
Meldonium Dihydrate*
Methane*
Permethrin*
Polyoxypropylene Stearyl Ether*
Pullulan*
Rosuvastatin Calcium*
Sulfadimethoxine*
Tizanidine Hydrochloride*
* denotes a monograph of the European Pharmacopoeia
I-xxxii Introduction 2016
Tolterodine Tartrate*
Zanamivir Hydrate*
Formulated Preparations: Specific Monographs
Abacavir and Lamivudine Tablets
Abacavir, Zidovudine and Lamivudine Tablets
Gastro-resistant Acamprosate Tablets
Bendroflumethiazide Oral Suspension
Carvedilol Tablets
Ceftazidime Eye Drops
Clotrimazole Eye Drops
Clotrimazole Vaginal Tablets
Diamorphine Tablets
Estradiol Vaginal Tablets
Etynodiol Tablets
Fluconazole Capsules
Fluconazole Infusion
Fluconazole Oral Suspension
Flumetasone and Clioquinol Ear Drops
Flupentixol Tablets
Fluticasone and Salmeterol Pressurised Inhalation Powder, pre-dispensed
Fluticasone and Salmeterol Pressurised Inhalation, Suspension
Interferon Beta-la Injection
Ketoconazole Cream
Ketoconazole Shampoo
Lorazepam Oral Solution
Miconazole Eye Drops
Olanzapine Tablets
Orodispersible Olanzapine Tablets
Olmesartan Tablets
Soluble Prednisolone Tablets
Rizatriptan Tablets
Orodispersible Rizatriptan Tablets
Prolonged-release Sodium Valproate Capsules
Prolonged-release Sodium Valproate Tablets
Terbinafine Tablets
Vecuronium Bromide for Injection
Zidovudine Infusion
Herbal Drugs, Herbal Drug Preparations and Herbal Medicinal
Products
Agnus Castus Fruit Dry Extract*
Barbary Wolfberry Fruit*
Holy Basil Leaf
Nettle Root*
Phellodendron Amurense Bark
Phellodendron Chínense Bark
Materials for use in the Manufacture of Homoeopathic Preparations
Coated Homoeopathic Pillules*
Agaricus Phalloides for Homoeopathic Preparations*
Ignatia for Homoeopathic Preparations*
Nux-vomica for Homoeopathic Preparations*
* denotes a monograph of the European Pharmacopoeia
2016 Introduction I-xxxiii
Blood-related Products
Normal Immunoglobulin for Subcutaneous Administration*
Immunological Products
Influenza Vaccine (Live, Nasal)*
Radiopharmaceutical Preparations
Fluoroethyl-L-tyrosine (18F) Injection*
Omissions The following monographs of the British Pharmacopoeia 2015 are not
included in the British Pharmacopoeia 2016.
Medicinal and Pharmaceutical Substances
Amitriptyline Embonate
Carbenoxolone Sodium
Chloroform
Fosfestrol Sodium
Halibut-liver Oil
Nandrolone Phenylpropionate
Formulated Preparations: Specific Monographs
Aminoglutethimide Tablets
Chloroform Spirit
Chloroform and Morphine Tincture
Double Strength Chloroform Water
Codeine Linctus1
Paediatric Codeine Linctus1
Codergocrine Tablets
Desipramine Tablets
Diethylstilbestrol Pessaries
Dydrogesterone Tablets
Ergometrine Tablets
Fosfestrol Injection
Fosfestrol Tablets
Fructose Infusion
Glucose Irrigation Solution
Halibut-liver Oil Capsules
Isoconazole Pessaries
Menadiol Phosphate Injection
Methylcellulose Granules
Morphine and Atropine Injection
Neonatal Naloxone Injection
Nandrolone Phenylpropionate Injection
Orciprenaline Oral Solution
Orciprenaline Tablets
Paraffin Ointment
Protriptyline Tablets
Tretinoin Solution
Technical Changes The following monographs in the British Pharmacopoeia 2016 have been
technically amended since the publication of the British Pharmacopoeia
2015, or have had a significant editorial change. This list does not include
revised monographs of the European Pharmacopoeia. An indication of the
nature of the change or the section of the monograph that has been
changed is given in italic type in the right hand column.
Medicinal and Pharmaceutical Substances
Nicorandil Loss on drying —> Water; Assay
Phenelzine Sulfate Assay
Changes in Tide The following list gives the alterations in the titles of monographs of the
British Pharmacopoeia 2015 that have been retained in the British
Pharmacopoeia 2016.
Blood-related Products
Normal Immunoglobulin Normal Immunoglobulin for
Intramuscular Administration
General Notices
1-2 General Notices 2016
General Notices
Part I
The British Pharmacopoeia comprises the entire text within this publication. The
word ‘official3is used in the Pharmacopoeia to signify (of the Pharmacopoeia\ It
applies to any title, substance, preparation} method or statement included in the
general notices, monographs and appendices of the Pharmacopoeia. The
abbreviation for British Pharmacopoeia is BP.
Part II
The following général notices apply to the statements made in the monographs of
the British Pharmacopoeia other than those reproduced from the European
Pharmacopoeia and to the statements made in the Appendices of the British
Pharmacopoeia other than when a method, test or other matter described in an
appendix is invoked in a monograph reproduced from the European
Pharmacopoeia.
Official Standards The requirements stated in the monographs of the Pharmacopoeia apply to
articles that are intended for medicinal use but not necessarily to articles
that may be sold under the same name for other purposes. An article
intended for medicinal use that is described by means of an official title
must comply with the requirements of the relevant monograph. A
formulated preparation must comply throughout its assigned shelf-life
(period of validity). The subject of any other monograph must comply
throughout its period of use.
A monograph is to be construed in accordance with any general
monograph or notice or any appendix, note or other explanatory material
that is contained in this edition and that is applicable to that monograph.
All statements contained in the monographs, except where a specific general
notice indicates otherwise and with the exceptions given below, constitute
standards for the official articles. An article is not of pharmacopoeial quality
unless it complies with all of the requirements stated. This does not imply
that a manufacturer is obliged to perform all the tests in a monograph in
order to assess compliance with the Pharmacopoeia before release of a
product. The manufacturer may assure himself that a product is of
pharmacopoeial quality by other means, for example, from data derived
from validation studies of the manufacturing process, from in-process
controls or from a combination of the two. Parametric release in
appropriate circumstances is thus not precluded by the need to comply with
the Pharmacopoeia. The general notice on Assays and Tests indicates that
analytical methods other than those described in the Pharmacopoeia may be
employed for routine purposes.
Requirements in monographs have been framed to provide appropriate
limitation of potential impurities rather than to provide against all possible
impurities. Material found to contain an impurity not detectable by means
of the prescribed tests is not of pharmacopoeial quality if the nature or
amount of the impurity found is incompatible with good pharmaceutical
practice.
The status of any statement given under the headings Definition,
Production, Characteristics, Storage, Labelling or Action and use is defined
within the general notice relating to the relevant heading. In addition to any
exceptions indicated by one of the general notices referred to above, the
following parts of a monograph do not constitute standards: (a) a graphic or
molecular formula given at the beginning of a monograph; (b) a molecular
weight; (c) a Chemical Abstracts Service Registry Number; (d) any
information given at the end of a monograph concerning impurities known
to be limited by that monograph; (e) information in any annex to a
2016 General Notices 1-5
Definition of Terms Where the term ‘about’ is included in a monograph or test it should be
taken to mean approximately (fairly correct or accurate; near to the actual
value).
Where the term ‘corresponds’ is included in a monograph or test it
should be taken to mean similar or equivalent in character or quantity.
Where the term ‘similar’ is included in a monograph or test it should be
taken to mean alike though not necessarily identical.
Further qualifiers (such as numerical acceptance criteria) for the above
terms are not included in the BP. The acceptance criteria for any individual
case is set based on the range of results obtained from known reference
samples, the level of precision of the equipment or apparatus used and the
level of accuracy required for the particular application. The user should
determine the variability seen in his/her own laboratory and set in-house
acceptance criteria that he/she judges to be appropriate based on the local
operating conditions.
Weights and The metric system of weights and measures is employed; SI Units have
Measures generally been adopted. Metric measures are required to have been
graduated at 20 ° and all measurements involved in the analytical operations
of die Pharmacopoeia are intended, unless otherwise stated, to be made at
that temperature. Graduated glass apparatus used in analytical operations
should comply with Class A requirements of the appropriate International
Standard issued by the International Organization for Standardization. The
abbreviation for litre is ‘L’ throughout die Pharmacopoeia. In line with
European Dừeetive 80/181/EEC, the abbreviation T is also permitted for
use.
Atomic Weights The atomic weights adopted are the values given in the Table of Relative
Atomic Weights 2001 published by the International Union of Pure and
Applied Chemistry (Appendix XXV).
Constant Weight The term ‘constant weight’, used in relation to the process of drying or the
process of ignition, means that two consecutive weighings do not differ by
more than 0.5 mg, the second weighing being made after an additional
period of drying or ignition under the specified conditions appropriate to
die nature and quantity of the residue (1 hour is usually suitable).
Expression of The term ‘per cent’ or more usually the symbol .*.%■is used with one of four
Concentrations different meanings in the expression of concenưations according to
cừcumstances. In order that the meaning to be attached to the expression
in each instance is clear, the following notation is used:
Per cent w/w (% w/w) (percentage weight in weight) expresses the
number of grams of solute in 100 g of product.
Per cent w/v (% w/v) (percentage weight in volume) expresses the
number of grams of solute in 100 mL of product.
Per cent v/v (% v/v) (percentage volume in volume) expresses the
number of millilitres of solute in 100 mL of product.
Per cent v/w (% v/w) (percentage volume in weight) expresses the
number OĨ millilitres of solute in 100 g of product.
Usually the sưength of solutions of solids in liquids is expressed as
percentage weight in volume, of liquids in liquids as percentage volume in
volume and of gases in liquids as percentage weight in weight.
When the concentration of a solution is expressed as parts per million
(ppm), it means weight in weight, unless otherwise specified.
When the concentration of a solution is expressed as parts of dissolved
substance in parts of the solution, it means parts by weight (g) of a solid in
parts by volume (mL) of the final solution; or parts by volume (mL) of a
liquid in pans by volume (mL) of the final solution; or parts by weight (g)
of a gas in parts by weight (g) of the final solution.
2016 General Notices 1-7
Water Bath The term ‘water bath’ means a bath of boiling water, unless water at some
other temperature is indicated in the text. An alternative form of heating
may be employed providing that the required temperature is approximately
maintained but not exceeded.
Reagents The reagents required for the assays and tests of the Pharmacopoeia are
defined in appendices. The descriptions set out in the appendices do not
imply that the materials are suitable for use in medicine.
Indicators Indicators, the colours of which change over approximately the same range
of pH, may be substituted for one another but in the event of doubt or
dispute as to the equivalence of indicators for a particular purpose, the
indicator specified in the text is alone authoritative.
The quantity of an indicator solution appropriate for use in acid-base
titrations described in assays or tests is 0.1 mL unless otherwise stated in
the text.
Any solvent required in an assay or test in which an indicator is specified
is previously neutralised to the indicator, unless a blank test is prescribed.
Caution Statements A number of materials described in the monographs and some of the
reagents specified for use in the assays and tests of the Pharmacopoeia may
be injurious to health unless adequate precautions are taken. The principles
of good laboratory practice and the provisions of any appropriate
regulations such as those issued in the United Kingdom in accordance with
the Health and Safety at Work etc. Act 1974 should be observed at all times
in carrying out the assays and tests of the Pharmacopoeia.
Attention is drawn to particular hazards in certain monographs by means
of an italicised statement; the absence of such a statement should not
however be taken to mean that no hazard exists.
Titles Subsidiary tides, where included, have the same significance as the main
tides. An abbreviated title constructed in accordance with the directions
given in Appendix XXI A has the same significance as the main title.
Titles that are derived by the suitable inversion of words of a main or
subsidiary tide, with the addition of a preposition if appropriate, are also
official titles. Thus, the following are all official tides: Aspirin Tablets,
Tablets of Aspirin; Atropine Injection, Injection of Atropine.
A title of a formulated preparation that includes the full nonproprietary
name of the active ingredient or ingredients, where this is not included in ^
the title of the monograph, is also an official tide. For example, the title
Promethazine Hydrochloride Oral Solution has the same significance as
Promethazine Oral Solution and the title Brompheniramine Maleate Tablets
has the same significance as Brompheniramine Tablets.
Where the English tide at the head of a monograph in the European
Pharmacopoeia is different from that at the head of the text incorporated
into the British Pharmacopoeia, an Approved Synonym has been created on
the recommendation of the British Pharmacopoeia Commission. Approved
Synonyms have the same significance as the main title and are thus official
1-8 General Notices 2016
Production Statements given under the heading Production draw attention to particular
aspects of the manufacturing process but are not necessarily comprehensive.
They constitute mandatory instructions to manufacturers. They may relate,
for example, to source materials, to the manufacturing process itself and its
validation and control, to in-process testing or to testing that is to be
carried out by the manufacturer on the final product (bulk material or
dosage form) either on selected batches or on each batch prior to release.
These statements cannot necessarily be verified on a sample of the final
product by an independent analyst. The competent authority may establish
that the instructions have been followed, for example, by examination of
data received from the manufacturer, by inspection or by testing
appropriate samples.
The absence of a section on Production does not imply that attention to
features such as those referred to above is not required. A substance,
preparation or article described in a monograph of the Pharmacopoeia is to
be manufactured in accordance with the principles of good manufacturing
practice and in accordance with relevant international agreements and
supranational and national regulations governing medicinal products.
Where in the section under the heading Production a monograph on a
vaccine defines the characteristics of the vaccine strain to be used, any test
methods given for confirming these characteristics are provided as examples
of suitable methods. The use of these methods is not mandatory.
Additional statements concerning the production of formulated
preparations are given in the general notice on Manufacture of Formulated
Preparations.
Freshly and The direction, given under the heading Extemporaneous Preparation, that a
Recently Prepared preparation must be freshly prepared indicates that it must be made not
more than 24 hours before it is issued for use. The direction that a
preparation should be recendy prepared indicates that deterioration is likely
if the preparation is stored for longer than about 4 weeks at 15° to 25°.
Water The term water used without qualification in formulae for formulated
preparations means either potable water freshly drawn direct from the
public supply and suitable for drinking or freshly boiled and cooled Purified
2016 General Notices 1-11
Water. The latter should be used if the public supply is from a local storage
tank or if the potable water is unsuitable for a particular preparation.
Antimicrobial When the term ‘suitable antimicrobial preservative’ is used it is implied that
Preservatives the preparation concerned will be effectively preserved according to the
appropriate criteria applied and interpreted as described in the test for
efficacy of antimicrobial preservation (Appendix XVI C). In certain
monographs for formulated preparations defined by means of a full formula,
a specific antimicrobial agent or agents may be prescribed; suitable
alternatives may be substituted provided that their identity and
concentration are stated on the label.
Identification The tests described or referred to under the heading Identification are not
necessarily sufficient to establish absolute proof of identity. They provide a
means of verifying that the identity of the material being examined is in
accordance with the label on the container.
Unless otherwise prescribed, identification tests are carried out at a
temperature between 15° and 25°.
Reference spectra Where a monograph refers to an infrared reference
spectrum, this spectrum is provided in a separate section of the
Pharmacopoeia. A sample spectrum is considered to be concordant with a
reference spectrum if the transmission minima (absorption maxima) of the
principal bands in the sample correspond in position, relative intensities and
shape to those of the reference. Instrumentation software may be used to
calculate concordance with a previously recorded reference spectrum.
When tests for infrared absorption are applied to material extracted from
formulated preparations, strict concordance with the specified reference
spectrum may not always be possible, but nevertheless a close resemblance
between the spectrum of the extracted material and the specified reference
spectrum should be achieved.
Assays an d Tests The assays and tests described are the official methods upon which the
standards of the Pharmacopoeia depend. The analyst is not precluded from
employing alternative methods, including jnethods of micro-analysis, in any
assay or test if it is known that the method used will give a result of
equivalent accuracy. Local reference materials may be used for routine
analysis, provided that these are calibrated against the official reference
materials. In the event of doubt or dispute, the methods of analysis, the
reference materials and the reference spectra of the Pharmacopoeia are
alone authoritative.
Where the solvent used for a solution is not named, the solvent is
Purified Water.
Unless otherwise prescribed, the assays and tests are carried out at a
temperature between 15° and 25°.
A temperature in a test for Loss on drying, where no temperature range
is given, implies a range of ± 2 ° about the stated value.
Visual com parative tests, unless otherwise prescribed, are carried out
using identical tubes of colourless, transparent, neutral glass with a flat
base. The volumes of liquid prescribed are for use with tubes 16 mm in
internal diameter; tubes with a larger internal diameter may be used but the
volume of liquid examined must be increased so that the depth of liquid in
the tubes is not less than that obtained when the prescribed volume of
liquid and tubes 16 mm in internal diameter are used. Equal volumes of the
liquids to be compared are examined down the vertical axis of the tubes
against a white background or, if necessary, against a black background.
The examination is carried out in diffuse light.
Where a direction is given that an analytical operation is to be carried out
‘in subdued light’, precautions should be taken to avoid exposure to direct
sunlight or other strong light. Where a direction is given that an analytical
operation is to be carried out ‘protected from light’, precautions should be
taken to exclude actinic light by the use of low-actinic glassware, working in
a dark room or similar procedures.
For preparations other than those of fixed strength, the quantity to be
taken for an assay or test is usually expressed in terms of the active
ingredient. This means that the quantity of the active ingredient expected to
2016 General Notices 1-13
substance itself. Where necessary for clarification the terms in which the
limit is expressed are stated within the monograph.
In all cases where an impurity limit is given in parentheses, the figures
given are approximations for information only; conformity with the
requirements is determined on the basis of compliance or otherwise with
the stated test.
The use of a proprietary designation to identify a material used in an
assay or test does not imply that another equally suitable material may not
be used.
Biological Assays Methods of assay described as Suggested methods are not obligatory, but
and Tests when another method is used its precision must be not less than that
required for the Suggested method.
For those antibiotics for which the monograph specifies a microbiological
assay the potency requirement is expressed in the monograph in
International Units (IU) per milligram. The material is not of
pharmacopoeial quality if the upper fiducial limit of error is less than the
stated potency. For such antibiotics the required precision of the assay is
stated in the monograph in terms of the fiducial limits of error about the
estimated potency.
For other substances and preparations for which the monograph specifies
a biological assay, unless otherwise stated, the precision of the assay is such
that the fiducial limits of error, expressed as a percentage of the estimated
potency, are within a range not wider than that obtained by multiplying by
a factor of 10 the square roots of the limits given in the monograph for the
fiducial limits of error about the stated potency.
In all cases fiducial limits of error are based on a probability of 95%
(P = 0.95).
Where the biological assay is being used to ascertain the purity of the
material, the stated potency means the potency stated on the label in terms
of International Units (IU) or other Units per gram, per milligram or per
millilitre. When no such statement appears on the label, the stated potency
means the fixed or minimum potency required in the monograph. This
interpretation of stated potency applies in all cases except where the
m onograph specifically directs otherwise.
Where the biological assay is being used to determine the total activity in
the container, the stated potency means the total number of International
Units (IU) or other Units stated on the label or, if no such statement
appears, the total activity calculated in accordance with the instructions in
the monograph.
Wherever possible the primary standard used in an assay or test is the
respective International Standard or Reference Preparation established by
the World Health Organization for international use and the biological
activity is expressed in International Units (IU).
In other cases, where Units are referred to in an assay or test, the Unit
for a particular substance or preparation is, for the United Kingdom, the
specific biological activity contained in such an amount of the respective
primary standard as the appropriate international or national organisation
indicates. The necessary information is provided with the primary standard.
Unless otherwise directed, animals used in an assay or a test are healthy
animals, drawn from a uniform stock, that have not previously been treated
with any material that will interfere with the assay or test. Unless otherwise
stated, guinea-pigs weigh not less than 250 g or, when used in systemic
General Notices 1-15
toxicity tests, not less than 350 g. When used in skin tests they are white or
light coloured. Unless otherwise stated, mice weigh not less than 17 g and
not more than 22 g.
Certain of the biological assays and tests of the Pharmacopoeia are such
that in the United Kingdom they may be carried out only in accordance
with the Animals (Scientific Procedures) Act 1986. Instructions included in
such assays and tests in the Pharmacopoeia, with respect to the handling of
animals, are therefore confined to those concerned with the accuracy and
reproducibility of the assay or test.
Action an d Use The statements given under this heading in monographs are intended only
as information on the principal pharmacological actions or the uses of the
materials in medicine or pharmacy. It should not be assumed that the
substance has no other action or use. The statements are not intended to be
binding on prescribers or to limit their discretion.
C rude D rugs; Herbal and complementary medicines are classed as medicines under European
T raditional H erbal Directive 2001/83/EC as amended. It is emphasised that, although requirements
and C om plem entary for the quality of the material are provided in the monograph to assist the
M edicines registration scheme by the UK Licensing Authority, the British Pharmacopoeia
Commission has not assessed the safety or efficacy of the material in traditional
use.
Monograph Title For traditional herbal medicines, the monograph title
is a combination of the binomial name together with a description of use.
Monographs for the material that has not been processed (the herbal drug)
and the processed material (the herbal drug preparation) are published
where possible. To distinguish between the two, the word ‘Processed’ is
included in the relevant monograph title.
Definition Under the heading Definition, the botanical name together
with any synonym is given. Where appropriate, for material that has not
been processed, information on the collection/harvesting and/or treatment/
drying of the whole herbal drug may be given. For processed materials, the
method of processing, where appropriate, will normally be given in a
separate section.
Characteristics References to odour are included only where this is
highly characteristic. References to taste are not included.
Control methods Where applicable, the control methods to be used in
monographs are:
(a) macroscopical and microscopical descriptions and chemical/
chromatographic tests for identification
(b) tests for absence of any related species
(c) microbial test to assure microbial quality
(d) tests for inorganic impurities and non-specific purity tests, including
extractive tests, Sulfated ash and Heavy metals where appropriate
(e) test for Loss on drying or Water
(f) wherever possible, a method for assaying the active constituent(s) or
suitable marker constituent (s).
The macroscopical characteristics include those features that can be seen
by the unaided eye or by the use of a hand lens. When two species/
subspecies of the same plant are included in the Definition, individual
differences between the two are indicated where possible.
The description of the microscopical characteristics of the powdered drug
includes information on the dominant or the most specific characters.
Where it is considered to be an aid to identification, illustrations of the
powdered drug may be provided.
The following aspects are controlled by the general monograph for
Herbal Drugs: they are required to be free from moulds, insects, decay,
animal matter and animal excreta. Unless otherwise prescribed the amount
of foreign matter is not more than 2 % w/w. Microbial contamination should
be minimal.
1-18 General Notices 2016
Part III
Monographs and other texts of the European Pharmacopoeia that are incorporated
in thừ edition of the British Pharmacopoeia are governed by the general notices of
the European Pharmacopoeia; these are reproduced below.
GENERAL NOTICES OF THE EUROPEAN
PHARMACOPOEIA
1.1. GENERAL STATEMENTS
The General Notices apply to all monographs and other texts of the
European Pharmacopoeia.
The official texts of the European Pharmacopoeia are published in
English and French. Translations in other languages may be prepared by
die signatory States of die European Pharmacopoeia Convention. In case of
doubt or dispute, die English and French versions are alone authoritative.
In the texts of the European Pharmacopoeia, the word ‘Pharmacopoeia5
without qualification means the European Pharmacopoeia. The official
abbreviation Ph. Eur. may be used to indicate the European
Pharmacopoeia.
The use of the title or the subtitle of a monograph implies that the article
complies with die requirements of die relevant monograph. Such references
to monographs in die texts of the Pharmacopoeia are shown using die
monograph title and reference number in italics.
A preparation must comply throughout its period of validity; a distinct
period of validity and/or specifications for opened or broached containers
may be decided by the competent authority. The subject of any other
monograph must comply throughout its period of use. The period of
validity that is assigned to any given article and the time from which that
period is to be calculated are decided by the competent authority in light of
experimental results of stability studies.
Unless otherwise indicated in the General Notices or in the monographs,
statements in monographs constitute mandatory reqmrements. General
chapters become mandatory when referred to in a monograph, unless such
reference is made in a way that indicates that it is not die intention to make
the text referred to mandatory but rather to cite it for information.
The active substances, excipients, pharmaceutical preparations and other
articles described in the monographs are intended for human and veterinary
use (unless explicitly restricted to one of these uses).
Quality systems The quality standards represented by monographs are valid only where the
articles in question are produced within the framework of a suitable quality
system. The quality system must assure that the articles consistently meet
die requirements of die Pharmacopoeia.
Alternative methods The tests and assays described are the official methods upon which the
standards of die Pharmacopoeia are based. With the agreement of die
competent authority, alternative methods of analysis may be used for
conưol purposes, provided that the methods used enable an unequivocal
decision to be made as to whether compliance with the standards of the
2016 General Notices 1-21
Demonstration of (1) An article is not of Pharmacopoeia quality unless it complies with all
compliance with the the requirements stated in the monograph. This does not imply that
Pharmacopoeia performance of all the tests in a monograph is necessarily a prerequisite
for a manufacturer in assessing compliance with the Pharmacopoeia
before release of a product. The manufacturer may obtain assurance
that a product is of Pharmacopoeia quality on the basis of its design,
together with its control strategy and data derived, for example, from
validation studies of the manufacturing process.
(2) An enhanced approach to quality control could utilise process analytical
technology (PAT) and/or real-time release testing (including parametric
release) strategies as alternatives to end-product testing alone. Real-time
release testing in circumstances deemed appropriate by the competent
authority is thus not precluded by the need to comply with the
Pharmacopoeia.
(3) Reduction of animal testing: the European Pharmacopoeia is dedicated
to phasing out the use of animals for test purposes, in accordance with
the 3Rs (Replacement, Reduction, Refinement) set out in the European
Convention for the Protection of Vertebrate Animals used for
Experimental and Other Scientific Purposes. In demonstrating
compliance with the Pharmacopoeia as indicated above ( 1),
manufacturers may consider establishing additional systems to monitor
consistency of production. With the agreement of the competent
authority, the choice of tests performed to assess compliance with the
Pharmacopoeia when animal tests are prescribed is established in such
a way that animal usage is minimised as much as possible.
Grade of materials Certain materials that are the subject of a pharmacopoeial monograph may
exist in different grades suitable for different purposes. Unless otherwise
indicated in the monograph, the requirements apply to all grades of the
material. In some monographs, particularly those on excipients, a list of
functionality-related characteristics that are relevant to the use of the
substance may be appended to the monograph for information. Test
methods for determination of one or m ore of these characteristics may be
given, also for information.
Validation of The test methods given in monographs and general chapters have been
pharmac opoeial validated in accordance with accepted scientific practice and current
methods recommendations on analytical validation. Unless otherwise stated in the
monograph or general chapter, validation of the test methods by the analyst
is not required.
Conventional terms The term ‘competent authority’ means the national, supranational or
international body or organisation vested with the authority for making
decisions concerning the issue in question. It may, for example, be a
national pharmacopoeia authority, a licensing authority or an official control
laboratory.
The expression ‘unless otherwise justified and authorised’ means that the
requirements have to be met, unless the competent authority authorises a
modification or an exemption where justified in a particular case.
Statements containing the word ‘should’ are informative or advisory.
In certain monographs or other texts, the terms ‘suitable’ and
‘appropriate’ are used to describe a reagent, micro-organism, test method
etc.; if criteria for suitability are not described in the monograph, suitability
is demonstrated to the satisfaction of the competent authority.
Medicinal product (a) Any substance or combination of substances
presented as having properties for treating or preventing disease in human
beings and/or animals; or (b) any substance or combination of substances
that may be used in or administered to human beings and/or animals with a
view either to restoring, correcting or modifying physiological functions by
exerting a pharmacological, immunological or metabolic action, or to
making a medical diagnosis.
Herbal m edicinal product Any medicinal product, exclusively
containing as active ingredients one or more herbal drugs or one or more
herbal drug preparations, or one or more such herbal drugs in combination
with one or more such herbal drug preparations.
Active substance Any substance intended to be used in the manufacture
of a medicinal product and that, when so used, becomes an active
ingredient of the medicinal product. Such substances are intended to
furnish a pharmacological activity or other direct effect in the diagnosis,
cure, mitigation, treatment or prevention of disease, or to affect the
structure and function of the body.
Excipient (auxiliary substance). Any constituent of a medicinal product
that is not an active substance. Adjuvants, stabilisers, antimicrobial
preservatives, diluents, antioxidants, for example, are excipients.
Interchangeable Certain general chapters contain a statement that the text in question is
methods harmonised with the corresponding text of the Japanese Pharmacopoeia
and/or the United States Pharmacopeia and that these texts are
interchangeable. This implies that if a substance or preparation is found to
2016 General Notices 1-23
Apparatus and Volumetric glassware complies with Class A requirements of the appropriate
procedures International Standard issued by the International Organisation for
Standardisation.
Unless otherwise prescribed, analytical procedures are carried out at a
temperature between 15 °C and 25 °C.
Unless otherwise prescribed, comparative tests are carried out using
identical tubes of colourless, transparent, neutral glass with a flat base; the
volumes of liquid prescribed are for use with tubes having an internal
diameter of 16 mm, but tubes with a larger internal diameter may be used
provided the volume of liquid used is adjusted (2.1.5). Equal volumes of
the liquids to be compared are examined down the vertical axis of the tubes
against a white background, or if necessary against a black background. The
exam ination is carried out in diffuse light.
Any solvent required in a test or assay in which an indicator is to be used
is previously neutralised to the indicator, unless a blank test is prescribed.
1-24 General Notices 2016
Water-bath The term ‘water-bath’ means a bath of boiling water unless water at
another temperature is indicated. Other methods of heating may be
substituted provided the temperature is near to but not higher than 100 °C
or the indicated temperature.
Drying and ignition The terms ‘dried to constant mass’ and ‘ignited to constant mass’ mean
to constant mass that 2 consecutive weighings do not differ by more than 0.5 mg, the 2nd
weighing following an additional period of drying or of ignition respectively
appropriate to the nature and quantity of the residue.
Where drying is prescribed using one of the expressions ‘in a desiccator’
or ‘in vacuo’, it is carried out using the conditions described in chapter
2.2.32. Loss on drying.
Solvents Where the name of the solvent is not stated, the term ‘solution’ implies a
solution in water.
Where the use of water is specified or implied in the analytical
procedures described in the Pharmacopoeia or for the preparation of
reagents, water complying with the requirements of the monograph Purified
water (0008) is used, except that for many purposes the requirements for
bacterial endotoxins (Purified water in bulk) and microbial contamination
(Purified water in containers) are not relevant. The term ‘distilled water’
indicates purified water prepared by distillation.
The term ‘ethanol’ without qualification means anhydrous ethanol. The
term ‘alcohol’ without qualification means ethanol (96 per cent). Other
dilutions of ethanol are indicated by the term ‘ethanol’ or ‘alcohol’ followed
by a statement of the percentage by volume of ethanol (C2H 60 ) required.
Relative Atomic and The relative atomic mass (A r) or the relative molecular mass (Mr) is shown,
Molecular Masses as and where appropriate, at the beginning of each monograph. The relative
atomic and molecular masses and the molecular and graphic formulae do
not constitute analytical standards for the substances described.
Chemical Abstracts CAS registry numbers are included for information in monographs, where
Service (CAS) applicable, to provide convenient access to useful information for users.
Registry Number CAS Registry Number^ is a registered trademark of the American
Chemical Society.
Characters The statements under the heading Characters are not to be interpreted in a
strict sense and are not requirements.
Solubility In statements of solubility in the Characters section, the terms
used have the following significance, referred to a temperature between
15 °C and 25 °C.
Soluble from 10 to 30
The term ‘partly soluble’ is used to describe a mixture where only some
of the components dissolve. The term ‘miscible’ is used to describe a liquid
that is miscible in all proportions with the stated solvent.
Identification Scope The tests given in the Identification section are not designed to give
a full confirmation of the chemical structure or composition of the product;
they are intended to give confirmation, with an acceptable degree of
assurance, that the article conforms to the description on the label.
First and second identifications Certain monographs have
subdivisions entitled ‘First identification’ and ‘Second identification’. The
test or tests that constitute the ‘First identification’ may be used in all
circumstances. The test or tests that constitute the ‘Second identification’
may be used in pharmacies provided it can be demonstrated that the
substance or preparation is fully traceable to a batch certified to comply
with all the other requirements of the monograph.
Certain monographs give two or more sets of tests for the purpose of the
first identification, which are equivalent and may be used independently.
One or more of these sets usually contain a cross-reference to a test
prescribed in the Tests section of the monograph. It may be used to
simplify the work of the analyst carrying out the identification and the
prescribed tests. For example, one identification set cross-refers to a test for
enantiomeric purity while the other set gives a test for specific optical
rotation: the intended purpose of the two is the same, that is, verification
that the correct enantiomer is present.
Powdered herbal drugs Monographs on herbal drugs may contain
schematic drawings of the powdered drug. These drawings complement the
description given in the relevant identification test.
Tests and Assays Scope The requirements are not framed to take account of all possible
impurities. It is not to be presumed, for example, that an impurity that is
not detectable by means of the prescribed tests is tolerated if common sense
and good pharmaceutical practice require that it be absent. See also below
under Impurities.
Calculation Where the result of a test or assay is required to be
calculated with reference to the dried or anhydrous substance or on some
other specified basis, the determination of loss on drying, water content or
other property is carried out by the method prescribed in the relevant test
in the monograph. The words ‘dried substance’ or ‘anhydrous substance’
etc. appear in parentheses after the result.
Where a quantitative determination of a residual solvent is carried out
and a test for loss on drying is not carried out, the content of residual
solvent is taken into account for the calculation of the assay content of the
substance, the specific optical rotation and the specific absorbance. No
further indication is given in the specific monograph.
Limits The limits prescribed are based on data obtained in normal
analytical practice; they take account of normal analytical errors, of
acceptable variations in manufacture and compounding and of deterioration
to an extent considered acceptable. No further tolerances are to be applied
to the limits prescribed to determine whether the article being examined
complies with the requirements of the monograph.
In determining compliance with a numerical limit, the calculated result of
a test or assay is first rounded to the number of significant figures stated,
unless otherwise prescribed. The limits, regardless of whether the values are
1-28 General Notices 2016
Storage The information and recommendations given under the heading Storage do
not constitute a pharmacopoeial requirement but the competent authority
may specify particular storage conditions that must be met.
The articles described in the Pharmacopoeia are stored in such a way as
to prevent contamination and, as far as possible, deterioration. Where
special conditions of storage are recommended, including the type of
container (see section 1.3. General chapters) and limits of temperature, they
are stated in the monograph.
The following expressions are used in monographs under Storage with
the meaning shown.
In an airtight container Means that the product is stored in an airtight
container (3.2). Care is to be taken when the container is opened in a damp
atmosphere. A low moisture content may be maintained, if necessary, by
the use of a desiccant in the container provided that direct contact with the
product is avoided.
Protected fr o m light Means that the product is stored either in a
container made of a material that absorbs actinic light sufficiently to protect
the contents from change induced by such light, or in a container enclosed
2016 General Notices 1-29
Warnings Materials described in monographs and reagents specified for use in the
Pharmacopoeia may be injurious to health unless adequate precautions are
taken. The principles of good quality control laboratory practice and the
provisions of any appropriate regulations are to be observed at all times.
Attention is drawn to particular hazards in certain monographs by means of
a warning statement; absence of such a statement is not to be taken to
mean that no hazard exists.
Impurities A list of all known and potential impurities that have been shown to be
detected by the tests in a monograph may be given. See also chapter 5.10.
Control of impurities in substances for pharmaceutical use. The impurities are
designated by a letter or letters of the alphabet. Where a letter appears to be
missing, the impurity designated by this letter has been deleted from the list
during monograph development prior to publication or during monograph
revision.
Md Specific optical rotation PPb Parts per billion (micrograms per kilogram)
bp Boiling point ppm Parts per million (milligrams p er kilogram)
BRP Biological Reference Preparation R Substance or solution defined under
CRS Chemical Reference Substance 4. Reagents
j20 Rf Retardation factor (see chapter 2.2.46)
“20 Relative density
X W avelength R:t U sed in chrom atography to indicate the ratio
o f the distance travelled by a substance to the
HRS H erbal reference standard distance travelled by a reference substance
IU International U nit RV Substance used as a prim ary standard in
M M olarity volumetric analysis (chapter 4.2.1)
Mr Relative m olecular mass
L D 50 T h e statistically determ ined quantity o f a Lo/10 dose T h e largest quantity of a toxin that, in the
substance that, when administered by the conditions of the test, when mixed with 0.1 IU
specified route, may be expected to cause the o f antitoxin and adm inistered by the specified
death o f 50 p er cent of the test animals within route, does not cause symptoms of toxicity in
a given period the test animals within a given period
M LD M inim um lethal dose L f dose T h e quantity of toxin or toxoid that flocculates
in the shortest time with 1 IU o f antitoxin
L + /1 0 dose T h e smallest quantity of a toxin that, in the
conditions of the test, when mixed with 0.1 IU CCIDso T h e statistically determ ined quantity o f virus
of antitoxin and administered by the specified th at may be expected to infect 50 p er cent of
route, causes th e death of the test anim als the cell cultures to which it is added
within a given period
EID50 T h e statistically determ ined quantity o f virus
L + dose T he smallest quantity o f a toxin that, in the th at may be expected to infect 50 per cent of
conditions of th e test, when mixed with 1 IU fertilised eggs into which it is inoculated
of antitoxin an d adm inistered by the specified ID 50 T h e statistically determ ined quantity of a virus
route, causes the death of the test animals that may be expected to infect 50 per cent of
within a given period the animals into which it is inoculated
lr/100 dose T he smallest quantity o f a toxin that, in the T h e statistically determ ined dose of a vaccine
P D 50
conditions o f the test, when mixed with that, in the conditions of the test, may be
0.01 IU of antitoxin and injected expected to protect 50 per cent o f the animals
intracutaneously causes a characteristic against a challenge dose of the micro
reaction at the site o f injection within a organisms or toxins against which it is active
given period
EDsn T h e statistically determ ined dose of a vaccine
L p/10 dose T h e smallest quantity o f toxin that, in the that, in the conditions of the test, may be
conditions o f the test, when mixed w ith 0.1 IU expected to induce specific antibodies in
of antitoxin an d adm inistered by the specified 50 per cent o f the animals for the relevant
route, causes paralysis in the test anim als vaccine antigens
within a given period
PFU Pock-forming units or plaque-form ing units
SPF Specified-pathogen-free.
2016 General Notices 1-31
Collections of micro-organisms
A TCC American Type Culture Collection NCTC N ational Collection of Type Cultures
10801 University Boulevard C entral Public Health Laboratory
Manassas, Virginia 20110-2209, USA Colindale Avenue
London NW9 5HT, Great Britain
C.I.P. Collection de Bactéries de l’Institut Pasteur
B.P. 52, 25 rue du D octeur Roux NCYC N ational Collection of Yeast C ultures
75724 Paris Cedex 15, France A FRC Food Research Institute
Colney Lane
IM I International Mycological Institute
Norwich NR4 7UA, Great Britain
Bakeham Lane
Surrey T W 20 9TY, Great Britain N IT E Biological Resource Center
D epartm ent of Biotechnology
I.P . Collection Nationale de Culture de
N ational Institute o f Technology and
Microorganismes (C.N .C.M .)
Evaluation
Institut Pasteur
2-5-8 Kazusakamatari, Kisarazu-shi, Chiba,
25, rue du D octeur Roux
292-0818
75724 Paris Cedex 15, France
Japan
N C IM B N ational Collection of Industrial and M arine
S.S.I. Statens Serum Institut
Bacteria Ltd
80 Amager Boulevard, Copenhagen, D enm ark
23 St Machar Drive
Aberdeen AB2 1RY, Great Britain
N CPF N ational Collection of Pathogenic Fungi
London School of Hygiene and Tropical
Medicine
Keppel Street
London W C1E 7 H T , Great Britain
1 The definitions of the units used in the International System are given in the booklet "Le Systeme International d’Unites (SI)” published by
the Bureau International des Poids et Mesures, Pavillion de Breteuil, F-92310 Sevres.
1-32 General Notices 2016
t = T-To
g = 9.806 65 m • s ~ 2
Length I metre m The metre is the length of the path travelled by light in a vacuum during a time
interval of 1/299 792 458 of a second.
Mass m kilogram kg The kilogram is equal to the mass of the international prototype of the kilogram.
The second is the duration of 9 192 631 770 periods of the radiation corresponding
Time t second s to the transition between the two hyperfine levels of the ground state of the
caesium-133 atom.
Electric current I ampere A The ampere is that constant current which, maintained in two straight parallel
conductors of infinite length, of negligible circular cross-section and placed 1 metre
apart in vacuum would produce between these conductors a force equal to 2 x 10'7
newton per metre of length.
Thermodynamic T kelvin K The kelvin is the fraction 1/273.16 of the thermodynamic temperature of the triple
temperature point of water.
Amount of substance n mole mol The mole is the amount of substance of a system containing as many elementary
entities as there are atoms in 0.012 kilogram of carbon-12*.
Luminous intensity I, candela cd The candela is the luminous intensity in a given direction of a source emitting
monochromatic radiation with a frequency of 540 x 10IJ hertz and whose energy
intensity in that direction is 1/683 watt per steradian.
* When the mole is used, the elementary entities must be specified and may be atoms, molecules, ions, electrons, other particles or specified groups of
such particles.
2016 General Notices 1-33
Table 1.6.-2. - SI units used in the European Pharmacopoeia and equivalence with other units
Quantity Unit
Name Name Symbol Expression b SI Expression in other Conversion of other units into SI units
Symbol
base units SI units
Wave number V one per metre 1/m m '1
pm
3° 3"
Wavelength X micrometre
o o
nanometre nm
Density P kilogram per cubic kg/m3 kg-m"3 1 g/mL = 1 g/cm3 =■ 103 kg-m'3
metre
Velocity V metre per second m/s m-s'1
Name Symbol
day d 1 d = 24 h - 86 400 s
Relative retention W ith reference to abacavir (retention — total: n o t more than 5 times the area o f the principal peak
time = about 17 min): im purity D = about 0.8; in the chrom atogram obtained with reference solution (b)
im purity A = about 0.9. (0.5 per cent);
System suitability: reference solution (a): — disregard limit. 0.5 times the area o f the principal peak in
— resolution: minim um 1.5 betw een the peaks due to the chrom atogram obtained with reference solution (b)
impurities D and A; m inim um 1.5 between the peaks due (0.05 per cent).
to impurity A and abacavir. H eav y m e ta ls (2.4.8)
Limit. M aximum 20 ppm.
— impurity A: not m ore than 3 times the area o f the 12 m L of solution S complies with test A. Prepare the
principal peak in the chrom atogram obtained with reference solution using 2 m L of lead standard solution
reference solution (b) (0.3 p er cent). (1 ppm Pb) R.
Related substances W a te r (2.5.32)
Liquid chromatography (2.2.29). Prepare the solutions M aximum 0.5 p er cent, determined on 60.0 mg.
immediately before use and transfer them to low-adsorption, inert
S u lfa te d a s h (2.4.14)
glass vials.
M aximum 0.2 per cent, determined on 1.0 g.
Test solution Dissolve 25 mg o f the substance to be examined
in water R and dilute to 100.0 m l. with the same solvent. A SSAY
Sonicate until dissolution is complete. Dissolve 0.300 g in 50 m L of water R. T itrate with 0.1 M
Reference solution (a) Dissolve 2.5 mg o f abacavir for peak sodium hydroxide, determining the end-point
potentiometrically (2 . 2 . 2 0 ).
identification CRS (containing impurities B and D ) in
10.0 m L of water R. 1 m L of 0.1 M sodium hydroxide is equivalent to 33.54 mg of
Reference solution (b) D ilute 1.0 m L of the test solution to C 28H 38N 12O 6S.
100.0 m L with water R. D ilute 1.0 m L o f this solution to IM P U R IT IE S
10.0 m L w ith water R. Specified impurities: A , B.
Column: Other detectable impurities (the following substances would, if
— size. I = 0.15 m , 0 = 3.9 m m ; present at a sufficient level, be detected by one or other o f
— stationary phase: end-capped octadecylsQyl silica gelfor the tests in the monograph. They are limited by the general
chromatography R (5 pm); acceptance criterion for other/unspecified impurities and/or
— temperature: 30 °C. by the general m onograph Substances for pharmaceutical use
Mobile phase: (2034). It is therefore n o t necessary to identify these
— mobile phase A: dilute 0.5 m L of trifluoroacedc acid R in impurities for dem onstration o f compliance. See also 5.10.
1000 m L o f water R ; Control of impurities in substances for pharmaceutical use): C, D,
— mobile phase B: water R, methanol R (15:85 V!V)\ E, F.
NH2 CHARACTERS
Acacia is almost completely b u t very slowly soluble, after
HjNr x >N about 2 h, in twice its mass of water leaving only a very small
residue of vegetable particles; the liquid obtained is colourless
or yellowish, dense, viscous, adhesive, translucent and weakly
a d d to blue litmus paper. Acada is practically insoluble in
ethanol (96 per cent).
C. [(15j4R )-4-(2,6-diarnino-9/f-purin-9-yl)cyclopent-2-
IDENTIFICATION
enyl] m ethanol,
A. A cada occurs as yellowish-white, yellow or pale amber,
sometimes with a pinkish tint, friable, opaque, spheroidal,
oval or reniform pieces (tears) o f a diameter from about
1-3 cm, frequently with a cracked surface, easily broken into
irregular, whitish or slightly yellowish angular fragments with
conchoidal fracture and a glassy and transparent appearance.
^ nI^ " >
h2na n In the centre o f an unbroken tear there is sometimes a small
cavity.
H
"Oc OH
B. Reduce to a powder (355) (2.9.12). T he pow der is white
or yellowish-white. Examine under a microscope using a
D. [(1 i?34i?)-4 -[2-amino-6-(cyclopropylainino)-9/f-purin-9- 50 per cent V/V solution of glycerol R. T h e pow der shows the
yl] cyclopent-2-enyl] methanol, following diagnostic characters: angular, irregular, colourless,
transparent fragments. Only traces of starch or vegetable
tissues are visible. N o stratified membrane, is ap p arent
^N H C. Examine the chromatograms obtained in the test for
glucose and fructose.
Results T h e chromatogram obtained with the test solution
h2n^ n^ n shows 3 zones due to galactose, arabinose and rhamnose.
H—r ^ \ ^ H N o other im portant zocies are visible, particularly in the
upper p art o f the chromatogram.
D . Dissolve 1 g o f the powdered herbal drug (355) (2.9.12)
E. [(li?,35)-3-[2-amino-6-(cyclopropylamino)-9/f-purin-9- in 2 m L of water R by stirring frequently for 2 h. Add 2 m L
yl] cyclopentyl] methanol, o f ethanol (96 per cent) R. After shaking, a white, gelatinous
mucilage is formed which becomes fluid on adding 10 m L of
water R.
NH TESTS
Solution S
H2NJ¿C> N N
Dissolve 3.0 g o f the powdered herbal drug (355) (2.9.12) in
25 m L o f water R by stirring for 30 min. Allow to stand for
30 min and dilute to 30 m L with water R.
"O<>x 0H
h í‘ h 3c ch3
Insoluble matter
M aximum 0.5 per cent.
T o 5.0 g o f the powdered herbal drug (355) (2.9.12) add
F. 6-(cyclopropylamino)-9- [(li?,45)-4- [ [(1,1 - 100 m L o f water R and 14 m L of dilute hydrochloric add R,
dimethylethyl)oxy] methyl] cyclopent-2-enyl] -9jFí-purine-2- boil gently for 15 min, shaking frequently and filter while hot
amine. through a tared sintered-glass filter (2.1.2). W ash with hot
PhEur water R and dry at 100-105 °C. The residue weighs a
maximum o f 25 mg.
Glucose and fructose
Thin-layer chromatography (2.2.27).
Acacia ★★+ ★★ Test solution T o 0.100 g of the powdered herbal drug (355)
★ ★ (2.9.12) in a thick-walled centrifuge tube add 2 m L of a
(Ph. Ever, monograph 0307) ***** 100 g/L solution of trifluoroacetic acid R, shake vigorously to
dissolve the forming gel, stopper the tube and heat the
Action and use mixture at 120 °C for 1 h. Centrifuge the hydrolysate,
Bulk-forming laxative; excipient transfer the clear supernatant carefully into a 50 m L flask,
W hen Powdered Acacia is prescribed or demanded, material add 10 m L of water R and evaporate the solution to dryness
complying with the requirements below with the exception of under reduced pressure. To the resulting clear film add
Identification test A shall be dispensed or supplied. 0.1 m L o f water R and 0.9 m L of methanol R. Centrifuge to
separate the amorphous predpitate. Dilute the supernatant, if
PhEur. necessary, to 1 m L with methanol R.
DEFINITION Reference solution Dissolve 10 mg of arabinose R, 10 mg of
Air-hardened, gummy exudate flowing naturally from or galactose R, 10 mg of glucose R, 10 mg o f rhamnose R and
obtained by incision of the trunk and branches o f Acacia 10 mg of xylose R in 1 m L of water R and dilute to 10 m L
Senegal L. Willd. (syn. Senegalia Senegal (L.) Britton), other with methanol R.
species of Acacia of African origin and Acacia seyal Delile.
1-42 Acacia 2016
Plate TUG silica gel plate R. part of the monograph since they also represent mandatory quality
Mobile phase 16 g/L solution of sodium dihydrogen phosphate R, criteria. In such cases, a cross-reference to the tests described in the
butanol R , acetone R (10:40:50 VIV/V). mandatory part is included in the Functionality-related
Application 10 ^L as bands. characteristics section. Control of the characteristics can contribute
to the quality of a medicinal product by improving the consistency
Development A Over a path o f 10 cm. of the manufacturing process and the performance of the medicinal
Drying A In a current o f warm air for a few minutes. product during use. Where control methods are cited, they are
Development B Over a path o f 15 cm using the same mobile recognised as being suitable for the purpose, but other methods can
phase. also be used. Wherever results for a particular characteristic are
Drying B A t 110 °C for 10 m in. reported, the control method must be indicated.
Detection treat with amsaldehyde solution R and heat at 110 °C The following characteristic may be relevant for acacia used as a
for 10 min. viscosity-increasing agent andlor suspending agent in aqueous
Results T he chromatogram obtained with the reference preparations.
solution shows 5 clearly separated coloured zones due to Apparent viscosity
galactose (greyish-green or green), glucose (grey), arabinose D eterm ine the dynamic viscosity using a capillary viscometer
(yellowish-green), xylose (greenish-grey or yellowish-grey) (2.2.9) or a rotating viscometer (2.2.10) on a 100 g/L
and rham nose (yellowish-green), in order of increasing RF solution o f acacia (dried substance).
value. T he chrom atogram obtained with the test solution ___________________________________________________________ PhEur
shows no grey zone and no greyish-green zone between the
zones corresponding to galactose and arabinose in the
chrom atogram obtained with the reference solution.
S ta r c h , d e x trin a n d a g a r Spray-dried Acacia ******
T o 10 m L o f solution S previously boiled and cooled add
0.1 m L of 0.05 M iodine. N o blue or reddish-brown colour (Ph Eur monograph 0308) *
develops. PhEur___________________________________________________________
S te rc u lia g u m D E F IN IT IO N
A. Place 0.2 g of the powdered herbal drug (355) (2.9.12) in Spray-dried acacia is obtained from a solution of acacia.
a 10 m L ground-glass-stoppered cylinder graduated in-
CHARACTERS
0.1 m L . Add 10 m L o f ethanol (60 per cent VIV) R and
shake. Any gel form ed occupies a m aximum o f 1.5 mT- It dissolves completely and rapidly, after about 20 m in, in
twice its mass of water. T h e liquid obtained is colourless or
B. T o 1.0 g of the powdered herbal drug (355) (2.9.12) add
yellowish, dense, viscous, adhesive, translucent and weakly
100 m L of water R and shake. Add 0.1 m L of methyl red
a d d to blue litmus paper. Spray-dried acada is practically
solution R. N o t m ore than 5.0 m L o f 0.01 M sodium hydroxide
insoluble in ethanol (96 p er cent).
is required to change the colour o f the indicator.
ID E N T IF IC A T IO N
T a n n in s
T o 10 m L o f solution S add 0.1 m L o f ferric chloride A. Examined under a microscope, in ethanol (96 per cent) R,
solution R l. A gelatinous precipitate is formed, bu t neither the the powder is seen to consist predom inantly o f spheroidal
precipitate nor the liquid are dark blue. particles about 4-40 (jm in diameter, w ith a central cavity
containing 1 or several air-bubbles; a few m inute flat
T ra g a c a n th
fragments are p resen t Only traces o f starch granules are
Examine the chromatograms obtained in the test for glucose visible. N o vegetable tissue is seen.
and fructose.
B. Examine the chromatograms obtained in the test for
Results T he chrom atogram obtained with the test solution glucose and fructose.
shows no greenish-grey or yellowish-grey zone corresponding
to the zone of xylose in the chrom atogram obtained with the
Results T h e chrom atogram obtained with the test solution
shows 3 zones due to galactose, arabinose and rham nose.
reference solution.
N o other im portant zones are visible, particularly in the
L oss o n d ry in g (2.2.32) upper p art o f the chromatogram.
M axim um 15.0 p er cent, determ ined on 1.000 g o f the
C. Dissolve 1 g of the drug to be examined in 2 m L of
pow dered herbal drug (355) (2.9.12) by drying in an oven at
water R by stirring frequently for 20 min. A dd 2 m L of
105 °C.
ethanol (96 per cent) R. After shaking a white gelatinous
T o ta l a sh (2.4.16) m udlage is formed which becomes fluid on adding 10 m L of
M axim um 4.0 per cent. water R.
M ic ro b ia l c o n ta m in a tio n TESTS
T A M C : acceptance criterion 104 C FU /g (2.6.12).
S o lu tio n S
TY M C : acceptance criterion 104 C FU /g (2.6.12). Dissolve 3.0 g o f th e drug to be examined in 25 m L of
Absence o f Escherichia cod (2.6.13). water R by stirring for 10 min. Allow to stand for 20 m in and
Absence o f Salmonella (2.6.13). dilute to 30 m L with water R.
F U N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S G lu co se a n d fru c to se
Thin-layer chromatography (2.2.27).
This section provides information on characteristics that are
recognised as being relevant control parameters for one or more Test solution T o 0.100 g in a thick-walled centrifuge tube add
functions of the substance when used as an excipient (see chapter 2 m L o f a 100 g/L solution o f trifluoroacetic acid R, shake
5.15). Some of the characteristics described in the Functionality- vigorously to dissolve the forming gel, stopper the tube and
related characteristics section may also be present in the mandatory h eat the mixture at 120 °C for 1 h. Centrifuge the
hydrolysate, transfer the clear supernatant carefully, into a
2016 Acamprosate Calcium 1-43
★★*★★
and rham nose (yellowish-green), in order of increasing
Rp value. T he chromatogram obtained with the test solution Acamprosate Calcium ★ ★
shows n o grey zone and no greyish-green zone between the *****
(Ph Eur monograph 1585)
zones corresponding to galactose and arabinose in the
chrom atogram obtained w ith the reference solution.
S ta r c h , d e x trin a n d a g a r
T o 10 m l. of solution S previously boiled and cooled add Ca2+
0.1 m L of 0.05 M iodine. N o blue or reddish-brown colour J2
develops.
Sterculia gum C io H ^ C a N ^ A ^ 400.5 77337-73-6
A. Place 0.2 g in a 10 m L ground-glass-stoppered cylinder
graduated in 0.1 mL. Add 10 m L of ethanol A ctio n a n d u se
(60 per cent VIV) R and shake. Any gel formed occupies not Treatm ent o f alcoholism.
more than 1.5 mL.
P re p a r a tio n
B. T o 1.0 g add 100 m L o f water R and shake. Add 0.1 m L Gastro-resistant Acamprosate Tablets
o f methyl red solution R. N o t more than 5.0 m L of
0.01 M sodium hydroxide is required to change the colour of PhEur__________________________________
the indicator. D E F IN IT IO N
T a n n in s Calcium bis[3-(acetylamino)propane-l-sulfonate].
T o 10 m L of solution S add 0.1 m L o f ferric chloride C o n te n t
solution R l. A gelatinous precipitate is formed, b u t neither the 98.0 per cent to 102.0 per cent (dried substance).
precipitate nor the liquid shows a dark blue colour.
CHARACTERS
Tragacanth
A p p e a ra n c e
Examine the chromatograms obtained in the test for Glucose
White or almost white powder.
and fructose.
Results T h e chromatogram obtained with the test solution Solubility
shows no greenish-grey o r yellowish-grey zone corresponding Freely soluble in water, practically insoluble in ethanol
to the zone of xylose in the chromatogram obtained with the (96 per cent) and in methylene chloride.
reference solution. ID E N T IF IC A T IO N
L oss o n d ry in g (2.2.32) A. Infrared absorption spectrophotometry (2.2.24).
M axim um 10.0 per cent, determined on 1.000 g by drying in Comparison Ph. Eur. reference spectrum of acamprosate calcium.
an oven at 105 °C. B. It gives reaction (a) o f calcium (2.3.1).
T o ta l a s h C2.4.16) TESTS
M axim um 4.0 per c e n t
S o lu tio n S
M ic ro b ia l c o n ta m in a tio n Dissolve 5.0 g in carbon dioxide-free water R and dilute to
T A M C : acceptance criterion 104 CFU/g (2.6.12). 100 m L with the same solvent.
T Y M C : acceptance criterion 102 C FU/g (2.6.12). A p p e a ra n c e o f so lu tio n
Absence of Escherichia colt (2.6.13). Solution S is clear (2.2.1) and colourless (2 .2 .2 , Method II).
1-44 Acarbose 2016
B. (li?,4i?,55,6iQ-4,5,6-trihydroxy-2-
(hydroxymethyl)cydohex-2-enyl 4-0-[4,6-dideoxy-4-
[ [(1 S,4uR,55, 65)-4,5,6-trihydroxy-3-
(hydroxymethyl)cyclohex-2-enyI] am ino]-a-D -
glucopyranosyl] -^-D -glucopyranoside,
G. a-D-glucopyranosyl 0 -4,6-dideoxy-4-[[(15,4ß ,55,65)-
4j5j6-trihydroxy-3-(hydroxymethyl)cyclohex-2-
en yl]am in o]-a-D -gIu cop yran osyl-(l -> 4 )-0 -a -D -
glucopyranosyl-(l ->4)-0 -a-D-glucopyranoside (a -D -
glucopyranosyl a-acarboside).
G )T 0H
lo \ — * OH
and enantiomer
O and enantiomer
A N - [3-acetyl-4- [(2i?3)-oxiran-2-
ylmethoxy]phenyl]butanamide, H . NyN'-[(2-hydroxypropane-l,3-diyl)bis[oxy(3-acetyl-1,4-
phenylene)]]dibutanamide.
R2 H OH PhEur
° ^ X ^ N CH3
j and enantiomer
R1. CH3
Aceclofenac ★ ★
★ ★
B. R I = R2 = C O -C H 3: N - [3-acetyl-4-[(2i?5)-2-hydroxy-3-
[( 1-methylethyl) amino] propoxy]phenyl] acetamide (Ph Eicr monograph 1281) *****
(diacetolol),
o co 2h
D . R I = H , R2 = CO-CH3: 1- [5-amino-2-[(2&S)-2-
hydroxy-3-[( 1-methyiethyl)amino]propoxy]phenyI] ethanone,
E. R I = C O -C H 2-C H 2-C H 33 R2 = H : N-[4-[(2i?5)-2-
o a
hydroxy-3-
[( 1-methylethyl) amino] propoxy]phenyI] butanamide,
J. R I = CO-CH2-CH3, R2 = CO-CH3: Ar-[3-acetyl-4-
[(2i?5)-2-hydroxy-3- C16H13a 2N04 354.2 89796-99-6
[( 1-methylethyl) amino] propoxy] phenyl] propanamide,
K R I = R2 = C O -C H 2-C H 2-C H 3: N-[3-butanoyl-4- A ctio n a n d u se
[(2i?5)-2-hydroxy-3- Cyclo-oxygenase inhibitor; analgesic; anti-inflammatory.
[( 1-methylethyl) amino] propoxy]phenyI]butanamide,
PhEur_______________________________________________________
DEFINITION
[[[2-[(2,6-Dichlorophenyl)amino]phenyl]acetyl]oxy]acetic
ad d .
C o n te n t
99.0 per cent to 101.0 p er cent (dried substance).
CHARACTERS
C. N-(3-acetyl-4-hydroxyphenyI)butanamide, A p p e a ra n c e
W hite or almost white, crystalline powder.
S o lu b ility
Practically insoluble in water, freely soluble in acetone,
soluble in ethanol (96 p er cent).
ID E N T IF IC A T IO N
First identification B.
2016 Aceclofenac 1-49
L oss o n d ry in g (2.2.32)
o
M aximum 0.5 per cent, determ ined on 1.000 g by drying in
an oven at 105 °C.
S u lfa te d a s h (2.4.14)
M aximum 0.1 per cent, determ ined on 1.0 g.
A SSA Y
Dissolve 0.300 g in 40 m L o f methanol R. T itrate with 0.1 M
sodium hydroxide, determining the end-point E. ethyl [[[2-[(2,6-
potentiometrically (2 . 2 . 2 0 ). dichlorophenyl)amino]phenyl] acetyl] oxy] acetate (ethyl ester
1 m L of 0.1 M sodium hydroxide is equivalent to 35.42 mg o f o f aceclofenac),
C 16H 13CI2N O 4.
STORAGE o
Protected from light.
IM P U R IT IE S
Specified impurities A, B, C, D , E, F , G, H , I
F. benzyl [[[2-[(2,6-
dichlorophenyl)amino]phenyI]acetyl]oxy]acetate (benzyl ester
o f aceclofenac),
O
A [2-[(2,6-dichlorophenyi)amino]phenyI] acetic acid
(diclofenac).
G. [[[[[2-[(2,6-
dichlorophenyl)amino]phenyl]acetyI] oxy] acetyl] oxy]acetic
acid (acetic aceclofenac),
B. methyl [2-[(2j6-dichlorophenyl)amino]phenyl]acetate
(methyl ester of diclofenac),
o
^ Y ^ r 0' ^ o / v 0 ^ ' c ° 2H
• OCT^
1. 1-(2 ,6-dichlorophenyl)-1,3-dihydro-2/f-indol-2-one.
PhEur
D . methyl [[[2-[(2,6-
dichlorophenyl)amino]phenyl]acetyl]oxy]acetate (methyl ester
of aceclofenac),
2016 Acemetacin 1-51
★ ★ — temperature: 40 °C.
Acemetacin ★ ★ Mobile phase:
(Ph Eitr monograph 1686) ***** — mobile phase A: dissolve 1.0 g of potassium dihydrogen
phosphate R in 900 m L of water R, adjust to p H 6.5 with
1 M sodium hydroxide and dilute to 1000 m L with
water R,
— mobile phase B: acetonitrUefor chromatography R]
DEFINITION
[ [[ l-(4-Chlorobenzoyl)-5-methoxy-2-methyl-1H-indol-3- Flow rate 1.0 mTVmin.
yl] acetyl] oxy] acetic ad d . Detection Spectrophotom eter at 235 nm.
C o n te n t Injection 20 pL.
99.0 per cent to 101.0 per cent (dried substance). Identification of impurities:
CHARACTERS — use the chromatogram supplied with acemetacin
A p p e a ra n c e impurity mixture CRS and the chromatogram obtained
Yellow or greenish-yellow, crystalline powder. with reference solution (e) to identify the peaks due to
impurities C, D , E and F;
Solubility — use the chromatogram obtained with reference
Practically insoluble in water, soluble in acetone, slightly solution (b) to identify the peak due to impurity B.
soluble in anhydrous ethanol.
Relative retention W ith reference to acemetacin (retention
It shows polymorphism (5.9). tim e = about 15 min): impurity A = about 0.7;
IDENTIFICATION impurity B = about 0.9; impurity F = about 1.2;
Infrared absorption spectrophotometry (2.2.24). impurity C = about 1.3; impurity D = about 1.5;
Comparison acemetacin CRS. impurity E = about 2.2.
I f the spectra obtained in the solid state show differences, System suitability Reference solution (d):
dissolve the substance to be examined and the reference — peak-to-valley ratio: minimum 15, where Hp = htight
substance separately in acetone R, evaporate to dryness and above the baseline of the peak due to impurity B and
record new spectra using the residues. Hv = height above the baseline of the lowest point o f the
curve separating this peak from the peak due to
TESTS acemetacin.
R e la te d su b sta n c e s Limits:
Liquid chromatography (2.2.29). — correction factors: for the calculation of content, multiply
Test sohaion Dissolve 0.100 g of the substance to be the peak areas o f the following impurities by the
examined in acetonitrUe for chromatography R and dilute to corresponding correction factor: impurity C = 1.3;
20.0 m L with the same solvent. impurity D = 1.4; impurity F = 1.3;
Reference solution (a) Dilute 5.0 mT. of the test solution to — impurity E: no t more than 3 times the area of the
50.0 m L with acetonitrUe for chromatography R. Dilute 1.0 m L p rindpal peak in the chromatogram obtained with
o f this solution to 100.0 m L with acetonitrUe for reference solution (a) (0.3 per cent);
chromatography R. — impurity B: not more than the area of the mi-responding
Reference sohaion (b) Dissolve 5.0 mg of acemetacin peak in the chromatogram obtained with reference
impurity A CRS and 10.0 m g o f indometacin CRS solution (c) (0.2 per cent);
(impurity B) in acetonitrUe for chromatography R, and dilute to — impurity A: not more than the area of the corresponding
50.0 m L with the same solvent. peak in the chromatogram obtained with reference
solution (c) (0.1 per cent);
Reference solution (c) Dilute 1.0 m L o f reference solution (b)
— impurities C, D, F: for each impurity, not more than the
to 20.0 m L with acetomtrUe for chromatography R.
area o f the prindpal peak in th e chromatogram obtained
Reference sohaion (d) T o 1 m L of reference solution (b), add with reference solution (a) (0.1 per cent);
10 m L o f the test solution and dilute to 20 m L with — unspecified impurities: for each impurity, not more than the
acetomtrUefor chromatography R. area o f the prindpal peak in the chromatogram obtained
Reference sohaion (e) Dissolve the contents o f a vial of with reference solution (a) (0.10 per cent);
acemetacin impurity mixture CRS (containing impurities C, D , — total: n o t more than 4 times the area of the principal peak
E and F) in 1.0 m L o f the test solution. in the chromatogram obtained with reference solution (a)
Column: (0.4 per cent);
— size: I = 0.25 m , 0 = 4 mm; — disregard limit. 0.5 times the area of the prindpal peak in
— stationary phase: spherical end-capped octadecylsUyl siHca gel the chromatogram obtained w ith reference solution (a)
for chromatography R (5 pm); (0.05 p er cent).
1-52 Acenocoumarol 2016
H eav y m e ta ls (2.4.8)
M aximum 20 ppm.
Acenocoumarol
Solvent mixture methanol R, acetone R (10:90 V!V).
0.250 g complies with test H . Prepare the reference solution
using 0.5 m L o f lead standard sohaion (10 ppm Pb) R.
L oss o n d ry in g (2.2.32)
M aximum 0.5 per cent, determ ined on 1.000 g by drying in
an oven at 105 °C.
S u lfa te d a s h (2.4.14)
M aximum 0.1 per cent, determined on 1.0 g. and enantiomer
A SSAY
Dissolve 0.350 g in 20 m L o f acetone R and add 10 m L of c 19h 15n o 6 353.3 152-72-7
water R. T itrate with 0.1 M sodium hydroxide, determining the
end-point potentiometrically (2 . 2 . 2 0 ). A ctio n a n d use
Vitamin K epoxide reductase inhibitor; oral anticoagulant.
1 m L o f 0.1 M sodium hydroxide is equivalent to 41.58 mg
o f C 2iH i8C 1N 06. P re p a r a tio n
Acenocoumarol Tablets
STORA GE
Protected from light. D E F IN IT IO N
IM P U R IT IE S Acenocoumarol is (R.S)-4-hydroxy-3-( 1-p-nitrophenyl-3-
Specified impurities A, B, C, D , E, F oxobutyl)coumarin. It contains not less than 98.5% and not
more than 100.5% o f C 19H15N 0 6, calculated with reference
COjH to the dried substance.
C H A R A C T E R IS T IC S
An almost white to buff powder.
Practically insoluble in water and in ether, slightly soluble in
ethanol (96%). It dissolves in aqueous solutions of the alkali
hydroxides. It exhibits polymorphism.
A. 4-chlorobenzoic acid,
ID E N T IF IC A T IO N
T he infrared absorption spectrum, Appendix II A, is concordant
with the reference spectrum of acenocoumarol (RS 001). If the
R3
spectra are n o t concordant, dissolve 0.1 g o f the substance
being examined in 10 m L o f acetone and add water drop wise
until the solution becomes turbid. H eat on a w ater bath until
the solution is clear and allow to stand. Filter, wash the
crystals with a mixture o f equal volumes of acetone and water
and dry at 100° at a pressure o f 2 kPa for 30 minutes.
B. R1 = R2 = R3 = H : [l-(4-chlorobenzoyl)-5-methoxy-2- Prepare a new spectrum o f the residue.
methylindol-3-yl]acetic acid (indom etadn),
TESTS
C. R1 = Cl, R2 = H , R3 = C H 2 -C 0 2H: [[[l-(3,4-
C la rity a n d c o lo u r o f so lu tio n
dichlorobenzoyl)-5-methoxy-2-methyl-l.H'-indol-3- A. A 2.0% w/v solution in acetone is clear, Appendix IV A.
yl] acetyl] oxy] acetic acid,
B. T he absorbance of a 4-cm layer of a 2.0% w/v solution in
D. R1 = H , R2 = C (C H 3)3, R3 = C H 2 -C 0 2H : [[[1-
acetone at 460 nm is not more than 0.12, Appendix II B.
(4-chlorobenzoyl)-6-( 1,1 -dimethylethyl)-5-methoxy-2-methyl-
1H-indol-3-yl] acetyl] oxy] acetic ad d , C. A 2.0% w/v solution in 0.1m sodium hydroxide is dear,
Appendix IV A, and yellow.
E. R1 = R2 = H , R3 = C H 2 -C 0 -0 -C (C H 3) 3:
1,1 -dimethylethyl [[[1 -(4-chlorobenzoyl)-5-methoxy-2- L ig h t a b so rp tio n
methyl- l/i-indol-3-yl] acetyl] oxy]acetate, Absorbance of a 0.001% w/v solution in a mixture of
1 volume o f 1m hydrochloric add and 9 volumes o f methanol at
F. R1 = R2 = H , R3 = C H 2- C 0 -0 - C H 2- C 0 2H: [[[[[1-
the maximum at 306 nm, 0.50 to 0.54, calculated with
(4-chlorobenzoyl)-5-methoxy-2-methjd-l/i-indol-3-
reference to the dried substance, Appendix II B.
yl] acetyl] oxy] acetyl] oxy] acetic ad d .
R e la te d su b sta n c e s
__________________________________________________________ PhEur
Carry out the m ethod for thin-layer chromatography,
Appendix HI A, using the following solutions in acetone.
(1) 2.0% w/v of the substance being examined.
(2) 0.0020% w/v o f the substance being examined.
CHROMATOGRAPHIC CONDITIONS
(a) U se as the coating silica gd GF2 s4 -
(b) Use die mobile phase as described below.
(c) Apply 20 nL of each solution.
(d) Develop the plate to 15 cm.
2016 Acesulfame Potassium 1-53
(e) After removal o f the plate, allow it to dry in air and Reference solution (a) Dissolve 5 mg o f acesulfame
im m ediately examine under ultraviolet light (254 nm). potassium CRS in water R and dilute to 5 m L with the same
solvent.
M O BILE PHASE
20 volumes of glacial acetic acid, 50 volumes of cychhexane Reference solution (b) Dissolve 5 mg o f acesulfame
and 50 volumes o f cHchloromethane.
potassium CRS and 5 m g o f saccharin sodium R in water R and
dilute to 5 m L with the same solvent.
LIMITS
Plate cellulose for chromatography R as the coating substance.
Any secondary spot in the chromatogram obtained with
Mobile phase concentrated ammonia R, acetone R, ethyl acetate R
solution (1) is n o t more intense than the spot in the
(10:60:60 V/V/V).
chrom atogram obtained with solution (2) (0.1%).
Application 5 pL as bands.
L oss o n d ry in g
W hen dried to constant weight at 105°, loses not more than Development Twice over 2/3 of the plate.
0.5% o f its weight. U se 1 g. Drying In a current o f warm air.
S u lfa te d ash Detection Examine in ultraviolet light at 254 nm.
N o t m ore than 0.1% , Appendix IX A. System suitability: reference solution (b):
— the chromatogram shows 2 clearly separated zones.
A SSA Y
Dissolve 0 .6 g in 5 0 m L of acetone and titrate with
Results T he principal zone in the chromatogram obtained
with the test solution is similar in position and size to the
0 .1 m sodium hydroxide VS using bromothymol blue solution R3
prindpal zone in the chromatogram obtained with reference
as indicator. Repeat the operation without the substance
solution (a).
being examined. T h e difference between the titrations
represents the am ount of sodium hydroxide required. Each C. 0.5 m L o f solution S (see Tests) gives reaction (b) of
mT. o f 0 .1 m sodium hydroxide KS is equivalent to 3 5 .3 3 m g of potassium (2.3.1).
C 19H 15N CV TESTS
S o lu tio n S
Dissolve 10.0 g in carbon dioxide-free water R and dilute to
50 m L with the same solvent.
Acesulfame Potassium ****** A p p e a ra n c e o f so lu tio n
*+ +* Solution S is clear (2.2.1) and colourless (2.2.2, Method IT).
(Ph Eur monograph 1282) *
A cid ity o r alk a lin ity
T o 20 m L of solution S add 0.1 m L of bromothymol blue
solution R l. N o t m ore than 0.2 m L o f 0.01 M hydrochloric
acid or 0.01 M sodium hydroxide is required to change the
colour of the indicator.
Im p u rity A
Thin-layer chrom atography (2.2.27).
Test solution Dissolve 0.80 g of the substance to be examined
C4H4KNO4S 201.2 55589-62-3
in water R and dilute to 10 m L with the same solvent
A c tio n a n d use Reference solution (a) Dissolve 50 m g o f acetylacetamide R
Sweetening agent. (impurity A) in water R and dilute to 25 m L with the same
solvent. T o 5 m L o f the solution add 45 m L of water R and
PhEur________________________________________________________ __ dilute to 100 m L with methanol R.
D E F IN IT IO N Reference solution (b) T o 10 m L of reference solution (a) add
Potassium 6-methyl-1,2,3-oxathiazin-4-olate 2,2-dioxide. 1 m L o f the test solution and dilute to 20 m L with
C o n te n t methanol R.
99.0 per cent to 101.0 per cent (dried substance). Plate TLC silica gel plate R.
CHARACTERS Mobile phase water R, ethanol (96 per cent) R, ethyl acetate R
(2:15:74 V/VIV).
A p p e a ra n c e
W hite or almost white, crystalline powder or colourless Application 5 piL.
crystals. Development Over 2/3 o f the plate.
S o lu b ility Drying In air until the solvents are completely removed.
Soluble in water, very slightly soluble in acetone and in Detection Spray with phosphoric vanillin solution R and heat at
ethanol (96 per cent). 120 °C for about 10 min; examine in daylight
ID E N T IF IC A T IO N System suitability T h e chromatogram obtained with reference
First identification A , C. solution (a) shows a clearly visible spot and the
chromatogram obtained with reference solution (b) shows
Second identification B, C.
2 clearly separated spots.
A. Infrared absorption spectrophotometry (2.2.24).
Limit.
Comparison acesulfame potassium CRS. — impurity A: any spot due to im purity A is not more
B. Thin-layer chromatography (2.2.27). intense than the spot in the chromatogram obtained with
Test solution Dissolve 5 mg of the substance to be examined reference solution (a) (0.125 per cent).
in water R and dilute to 5 m L with the same solvent. I m p u rity B
Liquid chromatography (2.2.29).
1-54 Acetazolamide 2016
Specific absorbance at the absorption maximum 162 to 176 for — impurities A , B, C, D, E, F: for each impurity, not more
solution A; 570 to 620 for solution B. than 1.5 times the area of the principal peak in the
B. Infrared absorption spectrophotometry (2.2.24). chromatogram obtained with reference solution (a)
(0.15 per cent);
Comparison acetazolamide CRS.
— unspecified impurities: for each impurity, not more than the
If the spectra obtained in the solid state show differences, area o f the principal peak in th e chromatogram obtained
dissolve the substance to be examined and the reference with reference solution (a) (0.10 per cent);
substance separately in ethanol (96 per cent) R, evaporate to
— total: not m ore than 6 times the area o f the principal peak
dryness and record new spectra using the residues. in the chromatogram obtained with reference solution (a)
C. Introduce about 20 mg into a test-tube and add 4 m L of (0.6 per cent);
dilute hydrochloric add R and 0.2 g o f zinc powder R — disregard limit. 0.5 times the area of the principal peak in
Im mediately place a piece of lead acetate paper R over the the chromatogram obtained w ith reference solution (a)
m outh o f the tube. T he paper shows a brownish-black (0.05 per cent).
colour.
S u lfates (2.413)
D . Dissolve about 25 mg in a mixture o f 0.1 m L o f dilute M axim um 500 ppm.
sodium hydroxide sdurion R and 5 m L of water R Add 0.1 m L
T o 0.4 g add 20 m L o f distilled water R and dissolve by
o f copper sulfate solution R. A greenish-blue precipitate is
heating to boiling. Allow to cool w ith frequent shaking and
formed.
filter.
TESTS H eav y m e ta ls (2.48)
Appearance of solution M aximum 20 ppm.
T he solution is no t more opalescent than reference
1.0 g complies with test C. Prepare the reference solution
suspension II (2.2.1) and not more intensely coloured than
using 2 m L o f lead standard solution (10 ppm Pb) R.
reference solution Y 5 or BY5 (2.2.2, Method II).
L oss o n d ry in g (2.2.32)
Dissolve 1.0 g in 10 m L of 1 M sodium hydroxide.
M aximum 0.5 per cent, determined on 1.000 g by drying in
Related substances an oven at 105 °C.
Liquid chromatography (2.2.29).
S u lfa te d a sh (2.414)
Test solution Dissolve 40 mg o f die substance to be examined M aximum 0.1 per cent, determined on 1.0 g.
in the mobile phase and dilute to 100.0 m L with the mobile
phase. A SSAY
Reference solution (a) Dilute 1.0 m L o f the test solution to Dissolve 0.200 g in 25 m L of dimethylforrnamide R. T itrate
100.0 m L with the mobile phase. Dilute 1.0 m L o f this with 0.1 M ethanolic sodium hydroxide, determining the
end-point potentiometrically (2 . 2 . 2 0 ).
solution to 10.0 m L with the mobile phase.
Reference solution (b) Dissolve the contents o f a vial o f 1 m L of 0.1 M ethanolic sodium hydroxide is equivalent to
acetazolamide for system suitability CRS (containing 22.22 m g of C 4H 6N 4O 3S2.
impurities A, B, C , D , E and F) in 1.0 m L o f the mobile IM P U R IT IE S
phase. Specified impurities A, B, C, D , E, F
Cdumn: Other detectable impurities (the following substances would, if
— size: I = 0.15 m , 0 = 4.6 mm; present at a sufficient level, be detected by one o r other of
— stationary phase: end-capped propoxybenzene silica gel for the tests in the monograph. They are limited by the general
chromatography R (4 (im). acceptance criterion for other/unspecified impurities and/or
Mobile phase acetonitrüe for chromatography R, 6.8 g/L solution by the general monograph Substances for pharmaceutical use
of potassium dihydrogen phosphate R (10:90 V!V). (2034). It is therefore not necessary to identify these
Flow rate 1.0 mL/min. impurities for demonstration of compliance. See also 5.10.
Detection Spectrophotom eter at 265 nm. Control of impurities in substancesfor pharmaceutical use): G.
Injection 25 jil. H
Run time 3.5 times the retention time of acetazolamide.
Identification of impurities Use the chromatogram supplied
\
N -N
Y R1
with acetazolamide for system suitability CRS and the
chromatogram obtained with reference solution (b) to A. R1 = C O -C H 3, R2 = Cl: N -(5-chloro-l,3,4-thiadiazol-2-
identify the peaks due to impurities A, B, C, D , E and F. yl)acetamide,
Relative retention W ith reference to acetazolamide (retention B. R1 = C O -C H 3, R2 = H: 2V-(l,3,4-thiadiazol-2-
time = about 8 min): impurity E = about 0.3; yl)acetamide,
impurity D = about 0.4; impurity B = about 0.6;
C. R1 = C O -C H 3, R2 = SH: N-(5-sulfanyl-1,3,4-thiadiazol-
impurity C = about 1.4; impurity A = about 2.1;
2-yl)acetamide,
impurity F = about 2.6.
D . R 1 = H , R2 = S 0 2-N H 2: 5-am ino-1,3,4-thiadiazole-2-
System suitability: reference solution (b):
sulfonamide,
— resolution: minim um 2.0 between the peaks due to
impurities E and D . E. R1 = C O -C H 3, R2 = S 0 2-0 H : 5-acetamido-1,3,4-
thiadiazole-2-sulfonic ad d ,
Limits:
— correction factory, for the calculation o f content, multiply
the peak areas o f the following impurities by the
corresponding correction factor impurity B = 2.3;
impurity C = 2 . 6; impurity D = 1.6;
1-56 Acetic Acid 2016
liberated iodine with 0.1m sodium tkiosidfate F 5 using starch mucilage as indicator. N ot less than 1.0 m L of 0.1m sodium
mucilage as indicator. N ot less than 0 .2 m L o f 0.1m sodium tkiosidfate VS is required.
thiosulfate F 5 is required. R ead ily o x id isa b le im p u ritie s
R ead ily o x id isab le im p u ritie s T o 5 .0 m L add 2 0 m L o f water and 0 .2 m L of
T o 2 5 m L add 0 .2 m L of 0.02m potassium permanganate FiS 0.02m potassium permanganate KS and allow to stand for
and allow to stand for 1 minute. T he pink colour is not 1 minute. T h e pink colour is not entirely discharged.
entirely discharged. N o n -v o la tile m a t te r
N o n -v o latile m a t te r W hen evaporated to dryness and dried at 105°, leaves not
W hen evaporated to dryness and dried at 105°, leaves not more th an 0 .01 % w/w of residue.
more than 0 .01 % w/w o f residue.
ASSAY
A SSAY Weigh 5 g into a stopper flask containing 5 0 m L o f water and
A dd 30 m L of water to 20 g in a stopper flask and titrate titrate w ith 1m sodium hydroxide F 5 using phenolphthalein
with 1m sodium hydroxide F 5 using phenolphthalein solution R1 solution R1 as indicator. Each mL o f 1m sodium hydroxide F S
as indicator. E ach m L of 1m sodium hydroxide F 5 is is equivalent to 6 0 .0 5 mg o f C2H 4 0 2.
equivalent to 60.05 mg of C 2H 4O 2.
Acetone ★ ★
★ ★
Acetic Acid (33 per cent) *****
(Ph Eur monograph 0872)
Acetic A d d
P r e p a r a tio n 0
Acetic A d d (6 p er cent) X , CH;
H3C
D E F IN IT IO N
Acetic A d d (33 p er cent) contains not less than 32.5% and
n o t m ore than 33.5% w/w of acetic a d d , C 2H 4O 2. CjHeO 58.08 67-64-1
PhEir___
C H A R A C T E R IS T IC S
A clear, colourless liquid. D E F IN IT IO N
M isdble with water, with ethanol (96%) and with glycerol. Propanone.
ID E N T IF IC A T IO N CHARACTERS
A. Strongly addic, even when diluted fredy. A p p e a ra n c e
Volatile, clear, colourless liquid.
B. W hen neutralised, yields the reactions characteristic of
acetates, A ppendix VI. S o lu b ility
M isdble w ith w ater and with ethanol (96 per cent).
TESTS
W eig h t p e r m l . T h e vapour is flammable.
1.040 to 1.042 g, Appendix V G. ID E N T IF IC A T IO N
H eav y m e ta ls A. Relative density (see Tests).
Evaporate 10.0 m L to dryness and add 20 m L o f water. B. T o 1 m L, add 3 m L o f dilute sodium hydroxide solution R
12 m L of the resulting solution complies with limit test A for and 0.3 m L of a 25 g/L solution o f sodium nitroprusside R.
heavy metals, Appendix VII. Use lead standard solution An intense red colour is produced which becomes violet with
(1 ppm Pb) to prepare the standard (2 ppm). the addition of 3.5 m L o f acetic acid R.
Chloride C. T o 10 m L of a 0.1 per cent VIV solution o f the substance
Dilute 5.0 m L w ith suffident water to produce 100 m L to be examined in ethanol (50 per cent VIV) R, add 1 m L o f a
15 m L of the resulting solution complies with the limit test for 10 g/L solution o f nitrobenzaldehyde R in ethanol
chlorides, Appendix VII (70 ppm). (50 per cent VIV) R and 0.5 mL o f strong sodium hydroxide
S u lfate solution R. Allow to stand for about 2 min and addify with
12.5 m L of the solution used in the test for Chloride, diluted
acetic acid R. A greenish-blue colour is produced.
to 15 m L with water, complies with the limit test for sulfates, TESTS
Appendix VII (240 ppm). A p p e a ra n c e o f so lu tio n
A ldehydes T o 10 m L add 10 m L o f water R. T he solution is clear
Distil 15 m L T o the first 5 m L of the distillate add 10 m L (2.2.1) and colourless (2.2.2, Method II).
of a 5% w/v solution of mercury(n) chloride, make alkaline A c id ity o r a lk a lin ity
w ith 5m sodium hydroxide, allow to stand for 5 minutes and T o 5 m L add 5 m L of carbon dioxide-free water R, 0.15 m L of
make addic w ith 1m sulfuric acid. The solution shows not phenolphthalein solution R and 0.5 m L of 0.01 M sodium
m ore than a faint turbidity. hydroxide. T h e solution is pink. A dd 0.7 m L o f 0.01 M
F o rm ic a c id a n d o x id isab le im p u ritie s hydrochloric acid and 0.05 m L of methyl red solution R.
M ix 5 m L with 6 m L o f sulfuric acid and cool to 20°. T h e solution is red or orange.
A dd 2 m L of 0 .0 1 6 7 m potassium dickromate F5, allow to R e lativ e d e n sity (2.2.5)
stand for 1 m inute, add 25 m L of water and 1 m L of freshly 0.790 to 0.793.
prepared dilute potassium iodide solution and titrate the
liberated iodine w ith 0.1m sodium thiosulfate VS using starch
1-58 Acetylcholine Chloride 2016
same solvent.
Reference solution (c) Dissolve 20 mg of choline chloride R in C. NjN-dimethylmethanamine.
methanol R, add 0.4 m L of test solution (a) and dilute to PhEur
2.0 m L with methanol R.
Plate TLC silica gel plate R.
Mobile phase Mix 20 volumes of a 40 g/L solution o f
ammonium nitrate R, 20 volumes of methanol R and Acetylcysteine ★* * ★★
60 volumes of acetonârUe R. ★ ★
Application 5 nL as bands o f 10 mm by 2 mm. (Ph. Eur. monograph 0967) *****
Development Over 2/3 o f the plate.
ch 3
Detection Spray with potassium iodobismuthate solution R3.
System suitability T h e chromatogram obtained with reference
0={
H NH
solution (c) shows 2 clearly separated zones.
co 2h
Limits:
— any impurity: any zones in the chromatogram obtained
with test solution (a), apart from the principal zone, are C 5H 9N 03 S 163.2 616-91-1
n o t more intense than the principal zone in the
Action and use
chrom atogram obtained with reference solution (a)
Sulfydryl donor; antidote to paracetamol poisoning;
(1 per cent).
mucolytic.
T rim e th y la m in e
P r e p a r a tio n
Dissolve 0.1 g in 10 m L of sodium carbonate solution R and
Acetylcysteine eye drops
heat to boiling. N o vapours appear which turn red litmus
paper R blue. Acetylcysteine Injection
Loss o n d ry in g (2.2.52)
Acetyldigoxin ★■k+ic★
M aximum 1.0 per cent, determined on 1.000 g by drying in ★ ★
★ ★
an oven in vacuo at 70 °C for 3 h. fl}~Acetyldigoxin, Ph Eur monograph 2168)
S u lfated a s h (2.4.14)
Maximum 0.2 per cent, determined on 1.0 g.
ASSAY
Dissolve 0.140 g in 60 m L o f water R and add 10 m L of
dilute hydrochloric acid R. After cooling in iced water, add
10 m L of potassium iodide solution R and titrate with 0.05 M
iodine, using 1 m L of starch solution R as indicator.
1 m L of 0.05 M iodine is equivalent to 16.32 m g of
C5H 9 N 0 3S.
STORAGE
Protected from light.
IMPURITIES
Specified impurities A, B, C, D
H NH,
HOjC
COjH
H NH2 C43H660 15 823 5355-48-6
PhEur_______________
H NH,
DEFINITION
co 2h
3 P- [(4- O-Acetyl-2,6-dideoxy-P-D-n&o-hexopyranosyl- (1 -> 4)-
2,6-dideoxy-P-D-nZ>o-hexopyranosyl-(l->4)-2,6-dideoxy-P-D-
B. (2i^-2-am ino-3-sulfanylpropanoic a d d (L-cysteine), n&o-hexopyranosyl)oxy]-l 2P, 14-dihydroxy-5P-card-20(22)-
enolide.
ch 3
Content
0=\ 97.0 p er cent to 102.0 p er cent (dried substance).
H NH
ho 2c CHARACTERS
> r ^ S ' Sx— X 'COjH
Appearance
H NH
W hite or alm ost white powder.
°= <
ch 3 Solubility
Practically insoluble in water, sparingly soluble in methylene
C. (2i?,2/i?)-3>3 /-disulfanediylbis[2-(acetylamino)propanoic chloride, slightly soluble in ethanol (96 per cent).
add] (N ^'-diacetyl-L-cystine), IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
ch 3 Comparison ^-acetyldigoxin CRS.
0=< TESTS
H NH
h3C^ S p ecific o p tic a l ro ta tio n (2.2.7)
Yo COjH
+ 26.2 to + 28.2 (dried substance).
Dissolve 0.50 g in a mixture of equal volumes o f methanol R
and methylene chloride R and dilute to 25.0 m L with the same
D. (2i?)-2-(acetylamino)-3-(acetylsulfenyl)propanoic a d d m ixture o f solvents.
(2V,.S-diacetyl-L-cysteine).
Related substances
PhEur
L iquid chrom atography (2.2.29). Prepare the solutions
immediately before use.
Solvent mixture M ix equal volumes of methanol R2 and
acetomtrUe for chromatography R.
Test solution Dissolve 50.0 mg of the substance to be
examined in the solvent mixture and dilute to 100.0 m L w ith
the solvent m ixture.
Reference solution (a) Dissolve 10.0 m g o f /?-acetyldigoxin CRS
in the solvent m ixture and dilute to 20.0 m L with the solvent
mixture.
Reference solution (b) Dilute 1.0 m L of the test solution to
20.0 m L w ith th e solvent mixture. D ilute 1.0 m L of this
solution to 10.0 m L with the solvent mixture.
1-62 Acetyldigoxin 2016
C. 3ß, 12ßjl4-trihydroxy-5ß-card-20(22)-enolide
(digoxigenin),
CH3
F . 3 ß- [(3,4-0-diacetyl-2,6-dideoxy-ß-i>nk>-hexopyranosyl-
( 1-^4)-2,6-dideoxy-ß-D-rÄö-hexopyranosyl-(l ->4)-2j6-
dideoxy-ß-D-rÄo-hexopyranosyl) oxy] -12 ßj 14-dihydroxy-5 ß-
card-20 (22)-enolide (diacetyldigoxin),
D . 3ß-[(2j6-dideoxy-ß-D-rÄo-hexopyranosyl-(l^4)-2j6-
dideoxy-ß-D-rÄo-hexopyranosyl-(l-^4)-2j6-dideoxy-ß-D-rÄo-
hexopyranosyl)oxy]-14jl6ß-dihydroxy-5ß-card-20(22)-enolide
(gitoxin),
ch3
G . 3 ß- [(3-0-acetyl-2,6-dideoxy-ß-i>nZ>o-hexopyranosyl-
(1 -+4)-2j6-*dideoxy-ß-D-r&o-hexopyranosyl-(l -+4)-2,6-
dideoxy-ß-D-rÄo-hexopyranosyl) oxy] -14-hydroxy-5 ß-caid-
20 (22)-enolide (a-acetyldigitoxin)j
H . 3ß-[(4-0-acetyl-2j6-dideoxy-ß-i>nZ>o-hexopyranosyl-
(1 ^4)-2,6-dideoxy-ß-D-rÄo-hexopyranosyl-(l -+4)-2,6-
dideoxy-ß-D-r&o-hexopyranosyl) oxy] -14-hydroxy-5 ß-card-
20(22)-enolide (ß-acetyldigitoxm).
___________________ :______________________________________ PhEur
1-64 Acetyltryptophan 2016
c o 2h
A. (S)-2-amino-3-(lH-indol-3-yl)propanoic acid
(tryptophan), J. R = C H O H -C H 2-O H: (5)-2-amino-3-[2- [2,3-dihydroxy-1-
(l/f-indol-3-yl) propyl] - lH-indol-3-yl]propanoic add,
K. R = H : (5)-2-amino-3-[2-(1 //-indol-3-ylmethyl)-1H-
and epimer at C*
indol-3-yl] propanoic ad d .
CO2H
B. (S)-2-amino-3-[(3i?S)-3-hydroxy-2-oxo-2,3-dihydro-li/-
indol-3-yl]propanoic a d d (dioxyindolylalanine),
1-66 Acetyltyrosine 2016
"¿0
M axim um 20 ppm .
OH
In a separating funnel, dissolve 0.5 g in 10 m L of dilute
h 2n
hydrochloric add R. Shake with 3 quantities, each o f 10 m L,
of methyl isobutyl ketone R l, shaking for 3 m in each time.
T o the combined organic layers add 10 m L o f water R and
C 8H 11N 5O 3 225.2 592 7 7 -8 9 -3
shake for 3 m in. T h e aqueous layer complies with the test.
H eav y m e ta ls (2.4.8) A c tio n a n d u se
Maximum 10 ppm . Purine nucleoside analogue; antiviral (herpesviruses).
Dissolve 2.0 g in water R and dilute to 20 m L with the same P re p a r a tio n s
solvent. 12 m L o f the solution complies with test A. Prepare Aciclovir C ream
the reference solution using lead standard solution Aciclovir Eye O intm ent
(1 ppm Pb) R.
Acidovir Infusion
L oss o n d ry in g (2.2.32)
A d d o v ir Oral Suspension
M aximum 0.5 p er cent, determined on 1.000 g by drying in
an oven at 105 °C. Adclovir Tablets
Dispersible A dd o v ir Tablets
S u lfa te d a sh (2.4.14)
M aximum 0.1 per cent, determined on 1.0 g. P h E u r __________________________________________________________________________________________
B a c te ria l e n d o to x in s (2.6.14)
D E F IN IT IO N
Less than 25 IU /g, if intended for use in the manufacture of
2-Amino-9- [(2-hydroxyethoxy) methyl] -1,9-dihydro-6//-purin-
parenteral preparations without a further appropriate
6-one.
procedure for the removal of bacterial endotoxins.
C o n te n t
A SSAY 98.5 per cent to 101.0 per cent (anhydrous substance).
Dissolve 0.180 g in 50 m L o f carbon dioxide-free water R.
T itrate with 0.1 M sodium hydroxide, determining the CHARACTERS
end-point potentiometrically (2 . 2 . 2 0 ). A p p e a ra n c e ^
W hite or almost white, crystalline powder.
1 m L of 0.1 M sodium hydroxide is equivalent to 22.32 m g of
C n H 13N 0 4. S o lu b ility
Slightly soluble in water, very slightly soluble in ethanol
STORA GE
(96 per cent), practically insoluble in heptane. It dissolves in
Protected from light. If the substance is sterile, store in a dilute solutions o f mineral adds and alkali hydroxides.
sterile, airtight, tam per-proof container.
ID E N T IF IC A T IO N
IM P U R IT IE S
Infrared absorption spectrophotometry (2.2.24).
Specified impurities A
Comparison aciclovir CRS.
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of TESTS
the tests in the monograph. They are limited by the general A p p e a ra n c e o f so lu tio n
acceptance criterion for other/unspecified impurities and/or T he solution is clear (2.2.1) and n o t more intensely coloured
by the general monograph Substances for pharmaceutical use than reference solution Y7 (2.2.2, Method IT).
(2034). It is therefore not necessary to identify these Dissolve 0.25 g in a 4 g/L solution of sodium hydroxide R and
impurities for demonstration o f compliance. See also 5.10. dilute to 25 m L with the same solvent.
Control of impurities in substancesfor pharmaceutical use): B. . R e la te d su b sta n c e s
Liquid chromatography (2.2.29). Prepare the solutions
immediately before use.
Solvent mixture dimethyl sulfoxide R, water R (20:80 VIV).
Phosphate buffer solution pH 2.5 Dissolve 3.48 g of dipotassium
A. (26)-2-amino-3-(4-hydroxyphenyl)propanoic acid hydrogen phosphate R in 1000 m L of water R and adjust to
(tyrosine), p H 2.5 with phosphoric add R.
1-68 Aciclovir 2016
H2N
i V>
h N N
1 aV>
h3c ^ n^ n^ n
L. N - (9-acetyl-6-oxo-6,9-dihydro-1.ff-purin-2-yl) acetamide
o -O OH (2^9-diacetylguanine),
1 1 / VJ
H2N N N 0
/^°x P ~ich3
C. 2-am ino-7- [(2-hydroxyethoxy)methyl]-l ,7-dihydro-6/i-
purin-6-onej
h3c^X ^JL^JL^
n n n
H
o
M . 2- [ [2-(acetylamino)-6-oxo-1,6-dihydro-7H-puiin-7-
yl]methoxy] ethyl acetate,
h3c ^ X a JCn> ;
" nA
H
^ n
\ — 0'
N . unknown structure,
O. unknown structure,
F. N-[9-[(2-hydroxyethoxy)methyl]-6-oxo-6,9-dihydro-l/i- 0
HNX .
purin-2-yl] acetamide,
o h2n ^ lX > n^ n^ ^ oh
O
i iY> ^
/
h3c ^ n ^ n ^ - n j—f ( 3
ch
P. 2-amino-9-(2-hydroxyethyl)-1,9-dihydro-6/f-purin-6-onej
H \— o
0
G. 2- [ [2-(acetylamino)-6-oxo-1,6-dihydro-9H-purin-9-
yl]methoxy] ethyl acetate.
x YN > N OH
V
HN JLn O
+
i xN >
H2N
x V>
h -O
/ \
OH
N NH2
H2N
a N
I> N / -----\
ccSl
N N NH2
'—O O—’
J. 9 ,9 '- [ethylenebis(oxymethylene)]bis(2-amino-l,9-dihydro-
6//-purin-6-one),
R. 9 ,9 '-
[methylenebis(oxyethyleneoxymethylene)]bis(2-amino-l,9-
dihydro-6//-purin-6-one).
PhEur
K. 2,2'-(methylenediimino)bis[9-[(2-hydroxyethoxy)methyl]-
1,9-dihydro-6/i-purin-6-one],
1-70 Acitretin 2016
TESTS
Acitretin ******
Related substances
(Ph Eur monograph 1385) * Liquid chrom atography (2.2.29). Maintain the sampler at
4 °C.
Test solution (a) Dissolve 2 5 .0 m g o f the substance to be
examined in 5 m L of tetrahydrofuran R and dilute
immediately to 1 0 0 .0 m L with anhydrous ethanol R.
Test solution (b) Dilute 1 0 .0 m L o f test solution (a) to
2 5 .0 m l. with anhydrous ethanol R.
Reference solution (a) Dissolve 2 5 .0 m g o f acitretin CRS in
Cj i H ^ O s 326.4 55079-83-9 5 m L of tetrahydrofuran R and dilute immediately to
100.0 m L with anhydrous ethanol R. D ilute 10.0 m L o f this
A ctio n a n d u se solution to 2 5 .0 m l. with anhydrous ethanol R
Vitamin A analogue (retinoid); treatm ent o f psoriasis; Reference solution (b) Dissolve 1 .0 m g of tretinoin CRS in
ichthyosis; D arier’s disease. anhydrous ethanol R and dilute to 2 0 .0 m L with the same
P re p a r a tio n solvent. M ix 5 .0 m L o f this solution with 2 .5 m L o f
A dtretin Capsules reference solution (a) and dilute to 1 0 0 .0 m L with anhydrous
ethanol R
PhEtr___________________________________________________________
Reference solution (c) Dilute 2 .5 m L o f reference solution (a)
D E F IN IT IO N to 5 0 .0 m L with anhydrous ethanol R. Dilute 3 .0 m L o f this
(aU-£)-9-(4-Methoxy-2,3, 6-trimethylphenyl)-3,7 - solution to 2 0 .0 m l. with anhydrous ethanol R
dimethylnona-2j4j6j8-tetraenoic ad d . Column’.
C o n te n t — size I = 0 .2 5 m , 0 = 4 mm;
98.0 per cent to 102.0 per cent (dried substance). — stationary phase: microparticulate octadecylsQyl silica gel for
chromatography R (5 pm) with a specific surface area of
CHARACTERS 2 0 0 m 2/g, a pore size o f 15 n m and a carbon loading o f
A p p e a ra n c e 20 per cent;
Yellow or greenish-yellow, crystalline powder. — temperature: 25 °C.
S o lubility Mobile phase A 0 .3 per cent V/V solution o f glacial acetic
Practically insoluble in water, sparingly soluble in acid R in a mixture o f 8 volumes o f water R and 9 2 volumes
tetrahydrofuran, slightly soluble in acetone and in ethanol o f anhydrous ethanol R.
(96 per cent), very slightly soluble in cyclohexane. Flow rate 0 .6 mL/min.
It is sensitive to air, heat and light, espedally in solution. Detection Spectrophotom eter at 3 6 0 nm .
It shows polymorphism. Injection 10 pL o f test solution (a) and reference solutions (b)
Cany ota all operations as rapidly as possible and avoid exposure and (c).
to actinic light; use freshly prepared solutions. Run time 2 .5 times the retention time o f adtretin.
ID E N T IF IC A T IO N Retention time Impurity A = about 4 .8 min; tretinoin = about
First identification B 5 .2 m in; adtretin = about 6 .2 min; impurity B = about
Second identification A , C 10.2 min.
A. Ultraviolet and visible absorption spectrophotom etry System suitability: reference solution (b):
(2.2.25). — resolution: minim um 2.0 betw een the peaks due to
Test solution Dissolve 15.0 m g in 10 m L o f tetrahydrofuran R adtretin and tretinoin; if necessary, adjust the
and dilute immediately to 100.0 m L with the same solvent. concentration o f anhydrous ethanol R
D ilute 2.5 m L o f this solution to 100.0 m L with Limits:
tetrahydrofuran R — impurities A , B: for each im purity, n o t more than the area
o f the peak due to adtretin in the chrom atogram obtained
Spectral range 300-400 nm.
w ith reference solution (c) (0 .3 p er cent);
Absorption maximum A t 358 nm . — total: n o t more than the area o f the peak due to ad tretin
Specific absorbance at the absorption maximum 1350 to 1475. in the chrom atogram obtained with reference solution (b)
B. Infrared absorption spectrophotom etry (2.2.24). (1.0 per cent);
Preparation Discs. — disregard limit. 0.1 times the area o f the principal peak in
the chrom atogram obtained with reference solution (c).
Comparison acitretin CRS.
If the spectra obtained in the solid state show differences, Palladium
dissolve die substance to be examined and the reference M axim um 10 ppm.
substance separately in 2-propanol R heating under reflux, A tomic absorption spectrometry (2.2.23, Method I).
filter, evaporate to dryness and record new spectra using the Test solution Introduce 2 .0 g into a quartz beaker and add
residues. 3 m l. o f magnesium nitrate solution R. H eat in a muffle
C. Examine the chrom atograms obtained in the assay. furnace to 3 5 0 °C at a rate o f 4 0 °C/min to incinerate the
Results T he prindpal peak in the chrom atogram obtained content. Ignite at about 4 5 0 °C for 8 h and then at
5 5 0 ± 5 0 °C for a further hour. Dissolve the residue in a
with test solution (b) is similar in retention time to the
m ixture o f 0 .7 5 m L o f hydrochloric add R and 0 .2 5 m L o f
principal peak in the chrom atogram obtained with reference
solution (a).
nitric acid R, warming gently. Cool, then transfer the solution
2016 Adapalene 1-71
Reference solution Dissolve 0.163 g o f heavy magnesium oxide R (Ph. Eur. monograph 2445) *
in a mixture o f 0.5 m L of nitric acid R, 1.5 m L o f hydrochloric
acid R and 50 m L o f water R, add 2.0 m L o f palladium
standard solution (20 ppm Pd) R and dilute to 100.0 m L with
water R.
Source Palladium hollow-cathode lamp.
Wavelength 247.6 nm.
Atomisation device Air-acetylene flame.
H eavy m e ta ls {2.4.8)
M aximum 20 ppm. C ss H a A 412.5 J 06685-40-9
2.0 g complies with test C. Prepare the reference solution A ctio n a n d u se
vising 2 m l. o f lead standard solution (10 ppm Pb) R. Vitamin A analogue (retinoid); treatm ent of acne.
L oss o n d ry in g (2.2.32) P re p a r a tio n s
M aximum 0.5 per cent, determined on 1.000 g by drying Adapalene Cream
in vacuo at 100 °C for 4 h. Adapalene Gel
S u lfated a s h (2.4.14)
M aximum 0.1 per cent, determined on 1.0 g. Ph Eur____________ _____________________________________________
ASSAY DEFINITION
Cany out the assay protected from light, use amber volumetric 6-(4-Methoxy-3-tricyclo [3.3.1.13,7] dec-1-
flasks and prepare the solutions immediately before use. ylphenyI)naphthalene-2-carboxylic acid.
Liquid chromatography (2.2.29) as described in the test for C o n te n t
related substances with the following modifications. 98.0 per cent to 102.0 per cent (dried substance).
Injection T est solution (b) and reference solution (a). CHARACTERS
System suitability: A p p e a ra n c e
— repeatability: maximum relative standard deviation of White or almost white powder.
1.0 per cent after 6 injections o f reference solution (a); Solubility
if necessary, adjust the integration parameters.
Practically insoluble in water, sparingly soluble in
Calculate the percentage content of C 21H 26O 3 from the tetrahydrofuran, practically insoluble in ethanol
declared content of acitrean CRS. (96 per cent).
STORAGE ID E N T IF IC A T IO N
In an airtight container, protected from light, at a Infrared absorption spectrophotometry (2.2.24).
tem perature o f 2 °C to 8 °C. Comparison adapalene CRS.
I t is recom mended that the contents o f an opened container
TESTS
be used as soon as possible and any unused part be protected
by an atmosphere o f inert gas. A p p e a ra n c e o f solution
T h e solution is clear (2.2.1) and not more intensely coloured
IM P U R IT IE S than reference solution BY6 (2.2.2, Method II).
Specified impurities A, B. Dissolve 0.2 g in tetrahydrofuran R and dilute to 20 m l. with
the same solvent.
ch 3 ch3 ch 3
R e la te d su b sta n c e s
Liquid chromatography (2.2.29).
h3co
Solvent mixture tetrahydrofuran R, acetomtrUe R, water R
(20:37:43 V/V/V).
ch 3
Test solution (a) Dissolve 40.0 mg o f the substance to be
examined in 10 m L o f tetrahydrofuran R, add 7 m L of the
A. (2Zj4£,6£’,8£)-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-
solvent mixture and dilute to 20.0 m L with tetrahydrofuran R.
dim ethylnona-2,4,6,8-tetraenoic add.
Test solution (b) Dissolve 20.0 m g o f the substance to be
examined in 50 m L of tetrahydrofuran R, add 35 m l. of the
solvent mixture and dilute to 100.0 m L with
tetrahydrofuran R. Dilute 5.0 m L o f the solution to 50.0 m L
with the solvent mixture.
Reference solution (a) Dilute 1.0 m L o f test solution (a) to
10.0 m L with tetrahydrofuran R. D ilute 1.0 m l. o f this
solution to 100.0 m l, with the solvent mixture.
B. ethyl (all-£)-9-(4-methoxy-2,336-trimethylphenyl)-3,7-
Reference solution (b) Dissolve 2.4 m g of adapalene
dim ethylnona-2,4,6,8-tetraenoate.
impurity C CRS in 2 m L of tetrahydrofuran R and dilute to
—-________________________________________________________ PhEur 20.0 m L with the same solvent. Dilute 2.0 m L o f the
solution to 20.0 m L with the solvent mixture. T o 2.0 m L of
this solution add 2.0 m L of reference solution (a) and dilute
to 20.0 m l. with the solvent mixture.
1-72 Adapalene 2016
Reference solution (c) Dissolve the contents of a vial of — disregard Omit: 0.5 times the area of the principal peak in
adapalene for peak identification CRS (containing impurities A, the chrom atogram obtained w ith reference solution (a)
C and D ) in 0.5 m L o f tetrahydrqfuran R and dilute to (0.05 per cent).
1.0 m L with the solvent mixture. H eav y m e ta ls (2.4.8)
Reference solution (d) Dissolve 20.0 m g o f adapalene CRS in M axim um 20 ppm .
50 m L of tetrahydrqfuran R> add 35 m L o f the solvent 0.250 g complies with test G . Prepare the reference solution
m ixture and dilute to 100.0 m L with tetrahydrofuran R. u sin g 0.5 m L of lead standard solution (10 ppm Pb) R.
D ilute 5.0 m L of the solution to 50.0 m L with die solvent
L oss o n d ry in g (2.2.32)
mixture.
M axim um 0.5 p er cent, determ ined on 1.000 g by drying in
Column: an oven at 105 °C for 4 h.
— size. I = 0.25 m , 0 = 4.6 mm;
— stationary phase: end-capped phenylsQyl silica gel for S u lfa te d a s h ( 2.4.14)
chromatography R (5 tun) with a carbon loading of M axim um 0.1 per cent, determ ined on 1.0 g.
7.5 per cent; A SSA Y
— temperature: 30 °C. Liquid chromatography (2.2.29) as described in the test for
Mobile phase: related substances w ith the following modification.
— mobile phase A: glacial acetic add R, water R (0.1:100 VIV)\ Irtjection T est solution (b) and reference solution (d).
— mobile phase B: tetrahydrofuran R, acetomtrUe R
Calculate the percentage content o f adapalene from the
(35:65 VIV)\
declared content o f adapalene CRS.
IM P U R IT IE S
Time Mobile phase A Mobile phase B
(min)
Specified impurities A, C , D
(percent V/V) (per cent V/V)
0 -2 .5 50 50 Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other o f
23 - 40 50-» 28 50*72
the tests in the m onograph. T hey are limited by die general
40 - 42 28 72 acceptance criterion for other/unspecified impurities and/or
by the general m onograph Substances for pharmaceutical use
(2034). It is therefore n o t necessary to identify these
Flow rate 1 .2 mL/min. impurities for dem onstration o f compliance. See also 5.10.
Detection Spectrophotom eter at 2 7 0 nm . Control of impurities in substances for pharmaceutical use): B.
Injection 2 5 |iL of test solution (a) and reference solutions (a),
(b) and (c).
Identification of impurities Use the chrom atogram supplied
with adapalene for peak identification CRS and the
chrom atogram obtained with reference solution (c) to identify
the peaks due to impurities A, C and D.
Relative retention W ith reference to adapalene (retention
tim e = about 2 0 min): impurity A = about 0 .3 ; A. 2 ,2 /-binaphthalene- 6,6 '-dicarboxylic acid,
impurity C = about 0 .9 ; im purity D = about 1.9.
System suitability: reference solution (b):
— resolution: m in im u m 4 .5 between the peaks due to
impurity C and adapalene;
— signal-to-noise ratio: minimum 1 0 for the peak due to
im purity C.
Limits: COjH
— correction factors: for the calculation of content, multiply
the peak areas o f the following impurities by the B. 6-[3-(3-hydroxytricydo[3.3.1.1 3,7]dec-l-yl)-4-
corresponding correction facto r im purity A = 0 .7 ; methoxyphenyl] naphthalene- 2-carboxylic ad d ,
impurity C = 7; impurity D = 1.4;
- — impurity A: not m ore than 3 times die area o f the
principal peak in the chrom atogram obtained with
reference solution (a) (0 .3 p er cent);
— impurity D: n o t m ore than twice die area o f the principal
peak in the chrom atogram obtained with reference
solution (a) (0.2 per cent);
C. l-(2-m ethoxyphenyi)tricydo[3.3.1. l 3,7]decane,
— impurity C: n o t m ore than 1.5 times the area o f the
principal peak in die chrom atogram obtained with
reference solution (a) (0 .1 5 p er cent);
— unspecified impurities: for each impurity, n o t m ore than the
area o f the principal peak in the chrom atogram obtained
w ith reference solution (a) (0.10 per cent);
— total: n o t m ore th an 5 times the area o f the principal peak
in the chrom atogram obtained with reference solution (a) D . 1, 1 '- [4,4 '-bis(methoxy)biphenyl-3,3
(0 .5 per cent); diyl]bis(tricyclo [3.3.1.13’7] decane).
______________________________________________■ PhEut
2016 Adenine 1-73
D E F IN IT IO N
nh2
Hexanedioic ad d .
C o n te n t
CT> N
N N
H
99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
A p p e a ra n c e
A. 7H-purin-6-amine (adenine).
W hite or alm ost white, crystalline powder.
S o lu b ility
Sparingly soluble in water, soluble in boiling water, fredy
soluble in ethanol (96 per cent) and in methanol, soluble in
acetone.
ID E N T IF IC A T IO N
A. M d tin g point {2.2.14): 151 °C to 154 °C.
B. Infrared absorption spectrophotometry {2.2.24).
OH OH
Comparison adipic acid CRS.
F. l-^i>ribofuranosylpyrim idine-2j4(l//j3/i)-dione TESTS
(uridine), S o lu tio n S
Dissolve 5.0 g with heating in distilled water R and dilute to
50 m L with the same solvent. Allow to cool and to
crystallise. Filter through a sintered-glass filter (40) {2.1.2).
HN W ash the filter with distilled water R. Collect the filtrate and
the washings until a volume of 50 m L is obtained.
HO A p p e a ra n c e o f so lu tio n
T he solution is clear {2.2.1) and colourless (2.2.2,
Method II).
Dissolve 1.0 g in methanol R and dilute to 20 m l. with the
OH OH
same solvent.
V? OH oh
Reference solution (b) Dilute 1.0 m L o f the test solution to
100.0 m L with the mobile phase, dilute 1.0 m L o f the
solution to 10.0 m L with the mobile phase.
Column:
H . 2-amino-9-P-D-ribofuranosyl-1,9-dihydro-6H-purin-6-one — size. I = 0.125 m , 0 = 4.0 mm,
(guanosine). — stationary phase, spherical octadecylstiyl silica gel for
PhEur chromatography R (5 Jim) with a specific surface area o f
350 m 2/g and a pore size of 10 nm,
— temperature: 30 °C.
Mobile phase M ix 3 volumes of acetonitrüe R and 97 volumes
o f a 24.5 g/L solution o f dilute phosphoric acid R.
Flow rate 1 m L/m in.
Detection Spectrophotometer at 209 nm.
Injection 20 (iL.
1-76 Adrenaline 2016
chromatography R and dilute to 1000 m L with the same corresponding correction factor: impurity D = 0.7;
solvent (it is usually necessary to stir for at least 30 min to impurity E = 0.6;
achieve complete dissolution). Adjust to p H 2.8 with — impurities B, C, F: for each impurity, not more than twice
phosphoric acid R. the area of the principal peak in the chromatogram
Solvent mixture B acetomtrUe R l, solvent mixture A obtained with reference solution (a) (0.2 per cent);
(13:87 VIV). — impurities D, E: for each impurity, not more than the area
Test solution Dissolve 40 mg of the substance to be examined o f the principal peak in the chromatogram obtained with
reference solution (a) (0.1 p er cent);
in 5 m L o f 0.1 M hydrochloric add and dilute to 50.0 m L
with solvent mixture B.
— unspedfied impurities: for each impurity, not more than the
area o f the principal peak in the chromatogram obtained
Reference solution (a) Dilute 1.0 m L of the test solution to with reference solution (a) (0.10 per cent);
100.0 m L with solvent mixture B. Dilute 1.0 m L o f this
— total: not m ore than 5 times the area o f the principal peak
solution to 10.0 m L with solvent mixture B.
in the chromatogram obtained with reference solution (a)
Reference solution (b) Dissolve 1.5 mg of noradrenaline (0.5 per cent);
tartrate CRS (impurity B) and 1.5 m g of adrendone — disregard limit. 0.5 times the area of the principal peak in
hydrochloride R (impurity C) in solvent mixture B, add the chromatogram obtained with reference solution (a)
1.0 m L of the test solution and dilute to 100 m L with (0.05 per cent).
solvent mixture B.
Loss on drying (2.2.32)
Reference solution (c) Dissolve the contents o f a vial o f M axim um 0.5 per cent, determined on 1.000 g by drying
adrenaline impurity mixture CRS (containing impurities D over diphosphorus pentoxide R at a pressure not exceeding
and E) in 1.0 m L o f the blank solution. 0.7 kPa for 18 h.
Reference solution (d) Dissolve 4 m g o f adrenaline with Sulfated ash (2.4.14)
impurity F CRS in 0.5 m L of 0.1 M hydrochloric add and M axim um 0.1 per cent, determined on 1.0 g.
dilute to 5 m L with solvent mixture B.
Blank solution 0.1 M hydrochloric acid, solvent mixture B ASSAY
(1:9 VIV). Dissolve 0.150 g in 50 m L of anhydrous acetic acid R. T itrate
with 0 . 1 M perchloric add, determining the end-point
Column:
potentiometrically (2 . 2 .2 0 ).
— size. I = 0.10 m , 0 = 4.6 mm;
— stationary phase, end-capped octadecylsOyl silica gel for 1 m L of 0.1 M perchloric add is equivalent to 18.32 mg
chromatography R (3 |xm); o f C 9H 13N 0 3.
— temperature: 50 °C. STORAGE
Mobile phase: U nder nitrogen, protected from light.
— mobile phase A: acetomtrUe R l, solvent mixture A
IM P U R IT IE S
(5:95 VIV)-,
Specified impurities B, C , D , E, F
— mobile phase B: acetonitrUe R l, solvent mixture A
(45:55 VIV)-,
HO
Time Mobile phase A Mobile phase B
(min) (per cent V7V) (per cent V/V) HO
0 - 15 92*50 8*50
O
Flow rate 2.0 mL/min.
Detection Spectrophotom eter at 210 nm.
Injection 20 (lL.
Identification of impurities Use the chromatogram supplied
with adrenaline impurity mixture CRS and the chromatogram C . 1-(3,4-dihydroxyphenyl)-2-(methylamino) ethanone
obtained with reference solution (c) to identify the peaks due (adrenalone),
to impurities D and E; use the chromatogram supplied with
adrenaline with impurity F CRS and the chromatogram
obtained with reference solution (d) to identify the peak due
to im purity F.
Relative retendon W ith reference to adrenaline (retention
time = about 4 min): impurity F = about 0.2;
impurity B = about 0.8; impurity C = about 1.3;
impurity D = about 3.3; impurity E = about 3.7.
System suitability: reference solution (b): D . 4-[(li?)-2-(benzyImethylamino)-l-hydroxyethyl]benzene-
— resolution: m inim um 3.0 between the peaks due to 1,2-diol,
im purity B and adrenaline.
Limits:
— correction factors: for the calculation of content, multiply
the peak areas o f the following impurities by the
1-78 Adrenaline Acid Tartrate 2016
STORAGE IDENTIFICATION
In an airtight container, or preferably in a sealed tube under A. Examine under a microscope. W hen m ounted in 0.005 M
vacuum or u n d er an inert gas, protected from light. iodine, the strips or flakes are partly stained brownish-violet.
Magnified 100 times, they show the following diagnostic
characters: numerous m inute, colourless, ovoid or rounded
1-80 Air 2016
grains on an am orphous background; occasional brown, solution add 5 m L o f picric acid solution R. N o turbidity
round or ovoid spores with a reticulated surface, measuring appears within 10 min.
up to 60 nm, may be present. Reduce to a powder, if L o ss o n d ry in g (2.2.32)
necessary. T h e pow der is yellowish-white. Examine un d er a M axim um 20.0 per cent, determ ined on 1.000 g o f the
microscope using 0.005 M iodine. T h e powder presents pow dered herbal drug (355) (2.9.12) by drying in an oven at
angular fragments w ith num erous grains similar to those seen 105 °C.
in the strips and flakes; some o f the fragments are stained
T o ta l a s h (2.4.16)
brownish-violet.
M axim um 5.0 per c e n t
B. Dissolve 0.1 g with heating in 50 m L of vocaer R. Cool.
M ic ro b ia l c o n ta m in a tio n
T o 1 m L of the mucilage carefully add 3 m L of water R so as
T A M C : acceptance criterion 103 C FU /g (2.6.12).
to form 2 separate layers. A dd 0.1 m L o f 0.05 M iodine.
A dark brownish-violet colour appears at the interface. Mix. T Y M C : acceptance criterion 102 C FU /g (2.6.12).
T he liquid becomes pale yellow. Absence o f Escherichia coli (2.6.13).
C. H eat 5 m L o f the mucilage prepared for identification Absence o f Salmonella (2.6.13).
test B on a w ater-bath with 0.5 m L o f hydrochloric acid R for
L A B E L L IN G
30 min. A dd 1 m L o f barium chloride solution R l. A white
T h e label states th e swelling index.
turbidity develops within 30 min.
___________________________________________________________ PhEur
D. H eat 0.5 g with 50 m L o f water R on a water-bath until
dissolved. Only a few fragments rem ain insoluble. D uring
cooling, the solution gels between 35 °C and 30 °C. H eat the
gel thus obtained on a water-bath; it does not liquefy below
80 °C. Medical Air *****
TESTS
(Medicinal Air\ Ph Eur monograph 1238) *
Sw elling in d e x (2.8.4)
M inim um 10 and within 10 p er cent o f the value stated on
When Medical Air is intended for use in a room in which
magnetic resonance imaging (MRJ) is being performed, the
the label, determ ined on the pow dered herbal drug (355)
cylinder and fittings should be made from suitable non-
(2.9.12).
ferromagnetic materials and labelled accordingly.
In so lu b le m a t te r
PhEur___________________________________________________________
M aximum 1.0 per cent.
T o 5.00 g o f the powdered herbal drug (355) (2.9.12) add D E F IN IT IO N
100 m i. of water R and 14 m L o f dilute hydrochloric add R. Compressed am bient air.
Boil gendy for 15 m in with frequent stirring. Filter the hot C o n te n t
liquid through a tared, sintered-glass filter (160) (2 . 1 . 2 ), rinse 20.4 per cent VIV to 21.4 per cent VIV o f oxygen (O 2).
the filter with hot water R and dry at 100-105 °C.
CHARACTERS
T h e residue weighs a maximum o f 50 mg.
A p p e a ra n c e
G elatin Colourless gas.
T o 1.00 g add 100 m L of water R and heat on a water-bath
until dissolved. Allow to cool to 50 °C. T o 5 m L o f this S o lu b ility
A t 20 °C at a pressure of 101 kPa, 1 volume dissolves in
about 50 volumes of water.
t
Photomultiplier
t
Amplifier
ID E N T IF IC A T IO N IM P U R IT IE S
First identification C. A. C O 2: carbon dioxide,
Second identification A , B. B. S 0 2: sulfur dioxide,
A. In a conical flask containing the substance to be C. N O : nitrogen monoxide,
examined, place a glowing wood splinter. T he splinter D . N 0 2: nitrogen dioxide,
rem ains glowing. E. oil,
B. U se a gas burette (Figure 1238.-2) o f 25 m L capacity in F. CO: carbon monoxide,
the form of a cham ber in the middle o f which is a tube
G . H 20 : water.
graduated in 0.2 per cent between 19.0 per cent and
23.0 per cent, and isolated at each end by a tap with a Ph Eur
STORAGE
As a compressed gas in suitable containers complying w ith
the legal regulations or as a compressed gas supplied by a
pipe network, after mixing of the components.
L A B E L L IN G
T he label states the nominal content of 0 2 in per cent VIV.
IMPURITIES
A. H 20 : water.
__________________________________________________________ PhEur
★★*★★
Alanine ★ ★
(Ph Eitr monograph 0752) *****
H NH,
V
HjC COjH
C3H7N 0 2 89.1 5 6 -4 1 -7
A ctio n a n d u se
Amino ad d .
PhEur
D E F IN IT IO N
(2S)-2-Aminopropanoic add.
C o n te n t
98.5 per cent to 101.0 per cent (dried substance).
CHARACTERS
A p p e a ra n c e
W hite or almost white, crystalline powder or colourless
crystals.
S o lu b ility
F red y soluble in water, very slightly soluble in ethanol
(96 per cent).
ID E N T IF IC A T IO N
First identification A , B
Second identification A , C, D
A. Specific optical rotation (see Tests).
B. Infrared absorption spectrophotometry (2.2.24).
Comparison alanine CRS.
C. Thin-layer chromatography (2.2.27).
Test solution Dissolve 10 mg of the substance to be examined
in water R and dilute to 50 m L w ith the same solvent.
Reference solution. Dissolve 10 mg of alanine CRS in water R
and dilute to 50 m L with the same solvent.
Plate TLC silica gel plate R.
Figure 1684.-1,- Gas burette
Mobile phase glacial acetic add R, water R, butanol R
(20:20:60 V/V/V).
enters the burette. Allow to stand for 10 min without Application 5 |xL.
shaking. Read the level of the liquid meniscus on the Development Over 2/3 of the plate.
graduated part o f the burette. T his figure represents the
Drying In air.
percentage VIV o f oxygen. T h e value read is 95.0 per cent to
105.0 p er cent o f the nominal value. Detection Spray with ninhydrin solution R and heat at 105 °C
for 15 min.
C. It complies w ith the limits of the assay.
Results T h e p rindpal spot in the chromatogram obtained with
TESTS the T est solution is similar in position, colour and size to the
W a te r v a p o u r prindpal spot in the chromatogram obtained with the
M axim um 67 p p m VIV, determined using a water vapour reference solution.
detector tube (2 . 1 . 6 ).
1-84 Alanine 2016
D . Dissolve 0.5 g in a mixture o f 0.25 m L o f hydrochloric — total: maximum 0.5 per cent;
add R l, 0.5 m L of a 100 g/L solution of sodium nitrite R and — reporting threshold: 0.05 per cent.
1 m L of water R. Shake; gas is given off, Add 2 m L o f dilute C h lo rid e s (2.4.4)
sodium hydroxide solution R, followed by 0.25 m L o f iodinaxed M aximum 200 ppm.
potassium iodide solution R. After about 30 min, a yellow
Dilute 5 m L of solution S to 15 m L with water R.
precipitate is formed.
S u lfates (2.4.13)
TESTS M aximum 300 ppm.
S o lu tio n S
Dilute 10 m L of solution S to 15 m L with dxstHled water R.
Dissolve 2.5 g in distilled water R and dilute to 50 m L with
the same solvent. A m m onium
Amino a d d analysis (2.2.56) as described in the test for
A p p e a ra n c e o f so lu tio n
ninhydrin-positive substances with the following
T he solution is clear ('2.2.1) and not more intensely coloured
modifications.
than reference solution BY 6 (2.2.2, Method II).
Injection T est solution, reference solution (c) and blank
Dilute 10 m L of solution S to 20 m L with water R.
solution.
S pecific o p tic a l r o ta tio n (2.2.7) Limit:
+ 13.5 to + 15.5 (dried substance). — ammonium at 570 ran: not more than the area of the
Dissolve 2.50 g in hydrochloric add R l and dilute to 25.0 m L corresponding peak in the chrom atogram obtained with
with the same acid. reference solution (c) (0.02 per cent), taking into account
N in h y d rin -p o sitiv e su b sta n c e s the peak due to ammonium in the chrom atogram
Amino a d d analysis (2.2.56). F or analysis, use M ethod 1. obtained with the blank solution.
T he concentrations o f the test solution and the reference Ir o n (2.4.9)
solutions may be adapted according to the sensitivity of the M aximum 10 ppm.
equipm ent used. T h e concentrations o f all solutions are In a separating funnd, dissolve 1.0 g in 10 m L of dilute
adjusted so that the system suitability requirements described hydrochloric add R. Shake with 3 quantities, each of 10 mL,
in general chapter 2.2.46 are fulfilled, keeping the ratios of of methyl isobutyl ketone R l, shaking for 3 m in each time.
concentrations between all solutions as described. T o the combined organic layers add 10 m L of water R and
Solution A dilute hydrochloric acid R l or a sample preparation shake for 3 min. Use the aqueous layer.
buffer suitable for the apparatus used. H eav y m e ta ls (2.4.8)
Test solution Dissolve 30.0 m g o f the substance to be M aximum 10 ppm.
examined in solution A and dilute to 50.0 m L with Dissolve 2.0 g in water R and dilute to 20 m L with the same
solution A. solvent. 12 m L of the solution complies with test A. Prepare
Reference solution (a) D ilute 1.0 m L of the test solution to the reference solution using lead standard solution
100.0 m L with solution A Dilute 2.0 m L of this solution to (1 ppm Pb) R.
10.0 m L with solution A. L oss o n d ry in g (2.2.32)
Reference solution (b) Dissolve 30.0 mg of proUne R in Maximum 0.5 per cent, determined on 1.000 g by drying in
solution A and dilute to 100.0 m L with solution A. Dilute an oven at 105 °C.
1.0 m L of the solution to 250.0 m L with solution A. S u lfa te d a s h (2.4.14)
Reference solution (c) Dilute 6.0 m L o f ammonium standard Maximum 0.1 per cent, determined on 1.0 g.
solution (100 ppm NH 4 ) R to 50.0 m L with solution A. D ilute
A SSAY
1.0 m L of this solution to 100.0 m L with solution A.
Dissolve 80.0 mg in 3 mL o f anhydrous formic add R.
Reference solution (d) Dissolve 30 m g o f isoleucine R and Add 30 m l. of anhydrous acetic add R. T itrate with 0.1 M
30 m g of leudne R in solution A and dilute to 50.0 m L with
perchloric add, determining the end-point potentiometrically
solution A. Dilute 1.0 m L o f the solution to 200.0 m L with
(2.2.20).
solution A.
1 m L of 0.1 M perchloric acid is equivalent to 8.91 m g of
Blank solution Solution A.
c 3h 7n o 2.
Inject suitable, equal am ounts of the test, blank and reference
solutions into the amino a d d analyser. Run a program STO RA G E
suitable for the determination of physiological amino adds. Protected from light.
System suitability Reference solution (d): IM P U R IT IE S
— resolution: minimum 1.5 between the peaks due to Other detectable impurities (the following substances would, if
isoleucine and leucine. present at a suffident levd, be detected by one or other of
Calculation of percentage contents: the tests in the monograph. They are limited by the general
— for any ninhydrin-positive substance detected at 570 nm , acceptance criterion for other/unspecified impurities and/or
use the concentration of alanine in reference solution (a); by the general monograph Substances for pharmaceutical use
— for any ninhydrin-positive substance detected at 440 nm , (2034). It is therefore no t necessary to identify these
use the concentration o f proline in reference solution (b); impurities for demonstration of compliance. See also 5.10.
if a peak is above the reporting threshold at both Control of impurities in substances for pharmaceutical use): A , B.
wavelengths, use the result obtained at 570 nm for
quantification. H nh 2
Column:
h o 2c ' — size. I = 0.25 m , 0 = 4.6 mm;
— stationary phase’, spherical end-capped octadecylsUyl silica gel
B. (25)-2-am inopentanedioic ad d (glutamic ad d ). for chromatography R (5 pm) with a pore size of 10 nm
PhEur and a carbon loading o f 19 p er cent.
Mobile phase Mix 300 volumes of a 1.67 g/L solution of
ammonium dihydrogen phosphate R and 700 volumes of
methanol R.
★ ★ Flow rate 0.7 mL/min.
Albendazole ★ ★
Detection Spectrophotom eter at 254 nm.
(Ph Eur monograph 1386) *****
Injection 20 |iL.
ch 3 Run time 1.5 times the retention time of albendazole.
Relative retention W ith reference to albendazole:
-NH impurity D = about 0.40; impurities B and C = about 0.43;
impurity E = about 0.47; impurity F = about 0.57;
impurity A = about 0.80.
C 12H 15N 3O 2S 265.3 54965-21-8 System suitability", reference solution (b):
— resolution', m inim um 3.0 between the peaks due to
A ctio n a n d u se
albendazole and oxibendazole.
Benzimidazole antihelminthic.
Limits:
P re p a r a tio n s — impurities A, B, C, D, E, F: for each impurity, not more
Albendazole Oral Suspension than 1.5 times the area of the principal peak in the
Albendazole Oral Suspension with Minerals chromatogram obtained with reference solution (a)
(0.75 p er cent);
PhEir.
— total: n o t more than 3 times the area o f the prindpal peak
D E F IN IT IO N in the chrom atogram obtained with reference solution (a)
M ethyl [5-(propylsulfanyl)-1 /f-benzimidazol-2 -yl] carbamate. (1.5 p er cent);
C o n te n t — disregard limit: 0.1 times the area of the prindpal peak in
98.0 per cent to 102.0 per cent (dried substance). the chromatogram obtained with reference solution (a)
(0.05 p er cent).
CHARACTERS
Loss on drying (2.2.32)
A p p e a ra n c e
M aximum 0.5 p er cent, determined on 1.000 g by drying in
W hite or slightly yellowish powder.
an oven at 105 °C for 4 h.
S o lu b ility
Sulfated ash (2.4.14)
Practically insoluble in water, freely soluble in anhydrous
M aximum 0.2 p er cent, determ ined on 1.0 g.
formic a d d , very slightly soluble in methylene chloride,
practically insoluble in ethanol (96 p er cent). ASSAY
In order to avoid overheating during the titration, mix thoroughly
ID E N T IF IC A T IO N
throughout and stop the titration immediately after the end-point
Infrared absorption spectrophotometry (2.2.24).
has been reached.
Preparation Discs.
Dissolve 0.250 g in 3 m L of anhydrous formic acid R and add
Comparison albendazole CRS. 40 m L o f anhydrous acetic add R. Titrate with 0.1 M
TESTS perchloric add, determining the end-point potentiometrically
A p p e a ra n c e o f so lu tio n (2.2.20).
T he solution is clear (2.2.1) and not m ore intensely coloured 1 m L of 0.1 M perchloric add is equivalent to 26.53 mg
than reference solution BY 6 (2.2.2, Method II). OfCi2Hj5N302S.
Dissolve 0.10 g in a mixture of 1 volume o f anhydrous formic STORAGE
add R and 9 volumes o f methylene chloride R and dilute to Protected from light.
10 m L w ith the same mixture of solvents.
IMPURITIES
R e la te d s u b s ta n c e s
Specified impurities A, B, C , D , E, F.
liq u id chrom atography (2.2.29).
Test solution Dissolve 25.0 m g o f die substance to be
examined in 5 m L o f methanol R containing 1 per cent VIV
-NH,
o f sulfuric add R and dilute to 50.0 m L with the mobile
phase.
Reference solution (a) Dissolve 10.0 m g o f the substance to be
examined in 10 m L of methanol R containing 1 p er cent VIV A. R = S-CH2-CH2-CH3: 5-(propylsulfanyl)-IH -
o f sulfuric acid R and dilute to 100.0 m L with the mobile benzimidazol- 2-am ine,
phase. D ilute 0.5 m L of this solution to 20.0 m L with the D . R = SO 2-C H 2-C H 2-C H 3: 5-(propyisulfonyI)-lii-
mobile phase. benzimidazol- 2-amine,
Reference solution (b) Dissolve 50.0 m g o f the substance to be
examined and 50 m g o f oxibendazole CRS in 5 m L o f
methanol R c o n taining l per cent VIV o f sulfuric add R and
dilute to 100.0 m L with the mobile phase.
1-86 Alcuronium Chloride 2016
O CH,
Reference solution Dissolve 10 m g o f alcuronium chloride CRS
1 y -o '
in methanol R and dilute to 10 m l. with the same solvent.
> -N H
Plate TLC silica gel plate R.
Mobile phase Mix 15 volumes o f a 58.4 g/L solution of sodium
B. R = SO -C H 2-C H 2-C H 3: methyi [5-(propylsuLfinyl)~ÎH- chloride R, 35 volumes of dilute ammonia R2 and 50 volumes
benzimidazol-2-yl]carfaamate, of methanol R.
C. R = SO 2-C H 2-C H 2-C H 3: methyl [5-(propylsulfonyl)-1H- Application 10 |iL.
benzimida 7ol- 2-yl] carbamate, Development Over a path of 15 cm.
E. R = H: methyl ( l.ff-benzimidazol-2-yl) carbamate, Drying In air for 10 min.
F. R = S-C H 3: methyl [5-(methylsulfanyl)-1/i-benziinidazol- Detection Spray with 0.1 M ammonium and cerium nitrate.
2-yl] carbamate. Results T he principal spot in the chrom atogram obtained with
__________________________________________________________ PhEur the test solution is similar in position, colour and size to the
principal spot in the chromatogram obtained with the
reference solution.
C. It gives reaction (a) of chlorides (2.3.1).
★*★
Alcuronium Chloride ★ ★ TESTS
★ ★
S o lu tio n S
(Ph Eur monograph 1285) *****
Dissolve 0.250 g in carbon dioxide-free water R and dilute to
25.0 m L with the same solvent.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and n o t more intensely coloured
than reference solution Y6, BY 6 or B 6 (2.2.2, Method I).
A cid ity o r a lk alin ity
2 Cl T o 10 m L of solution S add 0.1 m L of methyl red solution R
and 0.2 m L of 0.01 M hydrochloric add T he solution is red.
Add 0.4 m L of 0.01 M sodium hydroxide. T he solution is
yellow.
Specific o p tic a l ro ta tio n (2.2.7)
—430 to —451 (anhydrous substance), determined on
solution S.
C44H 50C I2N 4O2 738 15180-03-7
P ro p a n -2 -o l (2.4.24, System A)
A ctio n a n d u se Maximum 1.0 per cent.
Non-depolarizing neuromuscular blocker. R e la te d su b s ta n c e s
Liquid chromatography (2.2.29).
PhEur______________________________________
Solvent mixture M ix 100 m L of methanol R, 200 m L o f
D E F IN IT IO N acetonitrile R and 200 m L of a 6.82 g/L solution of potassium
(lfl,3aS,10S,llaS,12J?,14aS,19aS,20bS,21S,22aS,23.E,26£)- dihydrogen phosphate R. Dissolve 1.09 g o f sodium
23,26-bis(2-Hydroxyethylidene)-l, 12-bis(prop-2-enyl)- laurylsulfonate for chromatography R in the mixture and adjust
2,3,11,1 la , 13,14,22,22a-octahydro-10tf,21/f-l, 21:10,12- the apparent p H to 8.0 with a 100 g/L solution of sodium
diethano-19aH ,20b/f- [1,5]diazocino [ 1,2,3-Zm: 5,6,7- hydroxide R
l'm '] dipyrrolo[2',3 '-d:2'',3":d ']dicaibazolediium dichloride
Test solution Dissolve 0.20 g o f the substance to be examined
(4 ,4 /-didesmethyl-4,4/-bis(prop-2-enyl)toxiferm I dichloride).
in the solvent mixture and dilute to 100.0 m L with the
C o n te n t solvent mixture.
98.0 per cent to 102.0 per cent (anhydrous substance).
Reference solution (a) Dilute 0.5 m L o f the test solution to
CHARACTERS 100.0 m L with the solvent mixture.
A p p e a ra n c e Reference solution (b) Dilute 4.0 m l, of reference solution (a)
W hite or slightly greyish-white, crystalline powder. to 10.0 m L with the solvent mixture.
S olubility Reference solution (c) Dilute 1.0 m L o f reference solution (a)
Freely soluble in water and in methanol, soluble in ethanol to 10.0 m L with the solvent mixture.
(96 per cent), practically insoluble in cyclohexane. Reference solution (d) T o 5.0 m L o f the test solution add
Carry out the identification, tests and assay as rapidly as possible 5.0 mg of aUylstrychnine bromide CRS, dissolve in the solvent
avoiding exposure to actinic light mixture and dilute to 100.0 m L with the solvent mixture.
ID E N T IF IC A T IO N Column:
First identification A , C — size. I = 0.25 m, 0 = 4 mm;
— stationary phase: octylsQyl silica gel for chromatography R
Second identification B, C
(5 jim).
A. Infrared absorption spectrophotometry (2.2.24).
Mobile phase Mix 200 m L o f methanol R, 400 m L of
Comparison alcuronium chloride CRS. acetonitrile R and 400 m L o f a 6.82 g/L solution o f potassium
B. Thin-layer chromatography (2.2.27). dihydrogen phosphate R Dissolve 2.18 g o f sodium
Test solution Dissolve 10 m g o f the substance to be exam ined laurylsulfonate for chromatography R in the mixture and adjust
in methanol R and dilute to 10 m L with the same solvent.
2016 Alfacalcidol 1-87
6 injections.
Calculate the percentage content of C 27H 44O 2 taking into
account the assigned content o f alfacaladol CRS and, if
necessary, the peak due to pre-alfacalddol.
STORAGE
U nder nitrogen, in an airtight container, protected from light,
at a tem perature of 2 °C to 8 °C.
2016 Alfadex 1-89
CHA RACTERS
Reference solution (c) Dissolve 25.0 m g of alfadex CRS in
water R and dilute to 25.0 m L with the same solvent.
A p p e a ra n c e
Column:
White or almost white, amorphous or crystalline powder.
— sizer. I = 0.25 m, 0 = 4.6 mm;
S o lu b ility — stationary phase:, octadecylsUyl silica gel for chromatography R
Freely soluble in water, slightly soluble in propylene glycol, (10 |im).
practically insoluble in anhydrous ethanol and in methylene
Mobile phase methanol R, water R (10:90 VIV).
chloride.
Flow rate 1.5 mL/min.
ID E N T IF IC A T IO N
Detection Differential refractometer.
A. Specific optical rotation (see Tests).
Equilibration W ith the mobile phase for about 3 h.
B. Examine the chromatograms obtained in the assay.
Injection 50 |JL of test solution (a) and reference solutions (a)
Results T he principal peak in the chromatogram obtained
and (b).
with test solution (b) is similar in retention time and size to
the principal peak in the chromatogram obtained with Run time 3.5 times the retention time of alfadex.
reference solution (c). Relative retention W ith reference to alfadex (retention
C. Dissolve 0.2 g in 2 m L of iodine solution R4 by warming time = about 10 min): impurity B = about 0.7;
on a water-bath, and allow to stand at room temperature; impurity A = about 2.2.
a yellowish-brown precipitate is formed. System suitability: reference solution (a):
— resolution: minimum 1.5 between the peaks due to
TESTS
impurity B and alfadex; if necessary, adjust the
S o lu tio n S concentration of methanol in the mobile phase.
Dissolve 1.000 g in carbon dioxide-free water R and dilute to
Limits:
100.0 m L with the same solvent
— impurities A , B: for each impurity, not more than
A p p e a ra n c e o f so lu tio n 0.5 times the area of the corresponding peak in the
Solution S is clear (2.2.1). chromatogram obtained with reference solution (b)
p H (2.2.3) (0.25 per cent);
5.0 to 8.0. — sum of impurities other than A and B: not more than
Mix 1 m L o f a 223.6 g/L solution o f potassium chloride R and 0.5 times the area of the peak due to alfadex in the
30 m L o f solution S. chromatogram obtained with reference solution (b)
(0.5 per cent).
S p ecific o p tic a l ro ta tio n (2.2.7)
+ 147 to + 152 (dried substance), determined on solution S. Heavy metals (2.4.8)
Maximum 10 ppm.
R e d u c in g s u g a rs
2.0 g complies with test C. Prepare the reference solution
M aximum 0.2 p e r cent.
using 2 m L o f lead standard solution (10 ppm Pb) R.
Test solution T o 1 m L o f solution S add 1 m L o f cupri-tartaric
solution R4. H eat on a water-bath for 10 min, cool to room
1-90 Alfentanil Hydrochloride 2016
L oss o n d ry in g (2.2.32) ★ ★
M axim um 11 per cent, determ ined on 1.000 g by drying in
Alfentanil Hydrochloride .★ ★
an oven at 120 °C for 2 h. (Ph. Eur. monograph 1062) *****
S u lfa te d a s h (2.4.14)
M axim um 0.1 per cent, determ ined on 1.0 g.
n « nv ch3
A SSA Y J. N -V
Liquid chromatography (2.2.29) as described in the test for HCI
related substances with the following modifications.
Injection T est solution (b) and reference solutions (a) and (c). ch3
System suitability:
— repeatability: maximum relative standard deviation o f
2.0 per cent for the peak due to alfadex after 5 injections C21H33CIN6O3 453.0 69049-06-5
o f reference solution (a).
A c tio n a n d u se
Calculate the percentage content o f [ O H i o C ^ from the Opioid receptor agonist; analgesic.
assigned content of alfadex CRS.
PhEur_______________________________
STO R A G E
In an airtight container. D E F IN IT IO N
N -[l- [2-(4-Ethyl-4,5-dihydro-5-oxo-1H -tetrazol-1-yl) ethyl] -4-
IM P U R IT IE S
(methoxymethyl)piperidin-4-yl]-N-phenylpropanamide
Specified impurities A, B hydrochloride.
OH HO
C o n te n t
98.5 per cent to 101.5 per cent (anhydrous substance).
CHARACTERS
A p p e a ra n c e
W hite or alm ost white powder.
S o lu b ility
F red y soluble in water, in ethanol (96 p er cent) and in
methanol.
mp: about 140 °C, with decomposition.
ID E N T IF IC A T IO N
A. Infrared absorption spectrophotom etry (2.2.24).
Comparison Ph. Eur. reference spectrum of alfentanil
hydrochloride.
A. cycloheptakis-(l-»4)-(a-D-glucopyranosyl) (betadex or B. Dissolve 50 mg in a mixture of 0.4 m L o f ammonia R and
cyclomaltoheptaose or ß-cyclodextrin), 2 m L of water R. Mix, allow to stand for 5 min and filter.
A ddify the filtrate with dilute nitric add R. It gives
reaction (a) of chlorides (2.3.1).
TESTS
A p p e a ra n c e o f so lu tio n
T he solution is clear (2.2.1) and colourless (2.2.2,
Method IT).
Dissolve 0.2 g in water R and dilute to 20 m L with the same
solvent.
R e la te d su b s ta n c e s
Liquid chrom atography (2.2.29).
Test solution Dissolve 0.100 g of the substance to be
examined in methanol R and dilute to 10.0 m L with die same
solvent.
Reference solution (a) In order to produce impurity E in situ,
dissolve 10 mg of the substance to be examined in 10.0 m L
o f dilute hydrochloric add R. H eat on a water-bath under a
B. cyclooctakis-(l -*4)-(a-D-glucopyranosyl) reflux condenser for 4 h. Neutralise with 10.0 m L o f dilute
(cydomaltooctaose or y-cydodextrin). sodium hydroxide solution R. Evaporate to dryness on a water-
bath. Cool and take up the residue in 10 m L o f methanol R.
PhEur
Filter.
Reference solution (b) Dilute 1.0 m L o f the test solution to
100.0 m L with methanol R. Dilute 5.0 m L o f this solution to
20.0 m L w ith methanol R.
Column:
— size. I = 0.1 m , 0 = 4.6 mm;
2016 Alfentanil Hydrochloride 1-91
r i^ x î r
Flow rate 1.5 mL/min. o
Detection Spectrophotom eter at 220 nm. ^C H 3
Equilibration W ith acetomtrSe R for at least 30 min and then
0 V e
with the mobile phase at the initial composition for at least
B . trans-N- [ 1- [2-(4-ethyl-4,5-dihydro-5-oxo- lH -tetrazol-1-
5 min.
yl)ethyl]-4-(methoxymethyl)piperidin-4-yl]-iV-
Irqection 10 jiL; inject methanol R as a blank. phenylpropanamide N-oxide,
Retention time Im purity E = about 6 min; alfentanil = about
7 min. Ar / x
I f NH
Identification of impurities Use the chromatogram obtained
with reference solution (a) to identify the peak due to
o *^0 - CH3
impurity E; disregard any other peak.
System suitability: reference solution (a):
— resolution: m in im u m 4.0 between the peaks due to C. N - [4-(methoxymethyl)piperidm-4-yl]-N-
alfentanil and impurity E; if necessary, adjust the phenylpropanamide,
concentration o f acetonitrile in the mobile phase or adjust
the time programme for the linear-gradient elution.
Limits:
— impurities A , B, C, D, E, F, G, H: for each impurity, not
m ore than the area of the principal peak in the
chrom atogram obtained with reference solution (b)
(0.25 p er cent); D . N -[l- [2-(4-ethyl-4,5-dihydro-5-oxo-1//-tetrazol-1-
— total: not more than twice the area of the principal peak in yl)ethyI]-4-(methoxymethyl)piperidm-4-yl]-N-
the chromatogram obtained with reference solution (b) phenylacetamide,
(0.5 per cent);
— disregard limit. 0.2 times the area of the principal peak in
the chrom atogram obtained with reference solution (b)
(0.05 p er cent); disregard any peak due to the blank.
W a te r (2.5.12)
3.0 per cent to 4.0 p er cent, determined on 0.500 g.
E. 1-ethyl-1,4-dihydro-4- [2-[ [4-(methoxymethyl)-4-
A SSA Y phenylamino] piperidin-1-yl] ethyl]-5/f-tetrazol-5-one,
Dissolve 0.350 g in 50 m L of a m ixture o f 1 volume of
ethanol (96 per cent) R and 4 volumes o f water R and add Ar / s ,
5.0 m L o f 0.01 M hydrochloric add. T itrate with 0.1 M 1 I N
nh2 TESTS
Chlorides
B. R = Cl: 2<hloro-637-dimethoxyquina2olin-4-amine3 M axim um 1.0 p er cent.
D . R = N (C H 3)-[C H 2]3-N H 2: N-(4-amino-637- T o 2.50 g add 50 m L of dilute nitric acid R, shake for 1 h
dimethoxyquinazolin-2-yl) -N-methylpropane-1j3-diamine3 and dilute to 100.0 m L with dilute nitric add R. Filter.
T o 50.0 m L of the filtrate add 10.0 m L of 0.1 M silver nitrate
E. R = N (C H 3)-[C H 2]3-N H -CO -H : AT-[3-[(4-amino-6,7-
and 5 m L of toluene R. Titrate with 0.1 M ammonium
dimethoxyquinazolin-2-yl)methylamino]propyi]formamide,
thiocyanate, using 2 m L o f ferric ammonium sulfate solution R2
as indicator and shaking vigorously towards the end-point.
H H. / - , 1 m L of 0.1 M silver nitrate is equivalent to 3.545 mg o f Cl.
H’ °Y ^Y V N'v ^ NY b J Heavy m etals (2.4.8)
h 3c o
M axim um 20 ppm .
NH2 and enantiomer 1.0 g complies w ith test F . Prepare the reference solution
using 2 m L o f lead standard solution (10 ppm Pb) R.
C . (2RS)-N-[3-[(4-amino-6,7-dimethoxyquinazolin-2-yl) Loss on drying (2.2.32)
amino] propyl] -N-meiiiyltetrahydrofuran-2-carfaoxam ide. M axim um 15.0 p er cent, determined on 0.1000 g by drying
in an oven at 105 °C for 4 h.
PhEur
Sulfated ash (2.4.14)
Maximum 8.0 p er cent (dried substance), determined on
0.100 g.
M icrobial contamination
Alginic Acid ?**% T A M C : acceptance criterion 102 C FU /g (2.6.12).
(Ph Eur monograph 0591) * Absence o f Escherichia coli (2.6.13).
Absence o f Salmonella (2.6.13).
Action and use
T reatm ent o f gastro-oesophageal reflux disease; excipient; ASSAY
thickening agent. T o 0.2500 g add 25 m L of footer R, 25.0 m L o f 0.1 M
sodium hydroxide and 0.2 m L of phenolphthalein solution R.
PhEur__________________________________________________________ T itrate with 0.1 M hydrochloric acid.
DEFINITION 1 m L of 0.1 M sodium hydroxide is equivalent to 4.502 m g of
M ixture of polyuronic acids [(C6H80 6) J composed of carboxyl groups (-C 0 2H ).
residues of p-m annuronic and L-guluronic acids, obtained FUNCTIONALITY-RELATED CHARACTERISTICS
mainly from algae belonging to the Phaeophyceae. A small This section provides information on characteristics that are
proportion o f the carboxyl groups may be neutralised. recognised as being relevant control parameters for one or more
Content functions of the substance when used as an excipient (see chapter
19.0 per cent to 25.0 p er cent of carboxyl groups (-C 0 2H) 5.15). This section is a non-mandatory part of the monograph
(dried substance). and it is not necessary to verify the characteristics to demonstrate
CHARACTERS compliance. Control of these characteristics can however contribute
to the quality of a medicinal product by improving the consistency
Appearance
of the manufacturing process and the performance of the medicinal
W hite or pale yellowish-brown, crystalline or amorphous
product during use. Where control methods are cited, they are
powder.
recognised as bang suitable for the purpose, but other methods can
Solubility also be used. Wherever results for a particular characteristic are
Very slightly soluble o r practically insoluble in ethanol reported, the control method must be indicated
(96 per cent), practically insoluble in organic solvents.
The following characteristics may be relevant for alginic add used
It swells in w ater b u t does not dissolve; it dissolves in
as disintegrant and/or binder.
solutions of alkali hydroxides.
1-94 Alimemazine 2016
P a rtic le -siz e d is trib u tio n (2.9.31 or 2.9.38). Dissolve 1.0 g in water R and dilute to 10 m L with the same
S e ttlin g v o lu m e solvent.
Place 75 m L o f water R in a 100 m L graduated cylinder and p H (2.2.3)
add 1.5 g of die substance to be examined in 0.5 g portions, 5.0 to 6.5. Carry out the test protected from light and use a
shaking vigorously after each addition. Dilute to 100.0 m L freshly prepared solution.
with water R and shake again until the substance is Dissolve 1.0 g in carbon dioxide-free water R and dilute to
homogeneously distributed. Allow to stand for 4 h and 50 m L with the same solvent.
determine the volume o f the settled mass.
Related substances
The following characteristic may be relevant for alginic acid used Liquid chromatography (2.2.29). Carry out the test protected
as gelling agent or viscosity-increasing agent. from light and use freshly prepared solutions.
A p p a re n t viscosity Solvent mixture acetomtrüe R, water R (20:80 VIV).
D eterm ine the dynamic viscosity using a rotating viscometer
Test solution Dissolve 35 m g o f the substance to be examined
(2.2.1Q).
in the solvent mixture and dilute to 100.0 m L with the
Prepare a 20 g/L suspension o f alginic a d d (dried substance) solvent mixture.
and add 0.1 M sodium hydroxide until a solution is obtained.
Reference solution (a) D ilute 1.0 m L o f the test solution to
__________________________________________________________ PhEur 100.0 m L with the solvent mixture. D ilute 1.0 m L of this
solution to 10.0 m L with the solvent mixture.
Reference solution (b) Dissolve 3.5 m g o f alimemazine for
system suitability CRS (containing impurities A, B and C) in
***** the solvent mixture and dilute to 10.0 m L with the solvent
Alimemazine Tartrate ★ ★ mixture.
(Alimemazine Hemitartrate Ph. Bur. monograph ***** Column:
2650) — size. I = 0.15 m , 0 = 4.6 mm;
— stationary phase, base-deactivated end-capped octadecylsüyl
silica gelfor chromatography R (3 |im );
H OH — temperature. 40 °C.
n' ^ v ^ ' n ' CH3 COjH Mobile phase acetomtrüe R, methanol R , 3.854 g/L solution o f
I I H i i and dnantiomer , HOî C
CH3 c h 3 ammonium acetate R (10:40:50 VÍVIV).
H OH
Flow rate 1.3 mL/min.
Detection Spectrophotom eter at 253 nm .
Injection 20 jiL.
C20H 25N 2O3S 373.5 4330-99-8
Run time Twice the retention time o f alimemazine.
A ctio n a n d u se Identification of impurities Use the chrom atogram supplied
Histamine H i, receptor antagonist; sedative with alimemazine for system suitability CRS and the
P re p a ra tio n s chromatogram obtained with reference solution (b) to
Paediatric Alimemazine Oral Solution identify the peaks due to impurities A, B and C.
Strong Paediatric Alimemazine Oral Solution Relative retention W ith reference to alimemazine (retention
Alimemazine Tablets time = about 27 min): impurity A = about 0.1;
impurity B = about 0.5; impurity C = about 1.4.
PhEur__________________________________________ System suitability: reference solution (b):
D E F IN IT IO N — resolution: minimum 5.0 between the peaks due to
(2RS)-N,N, 2-Trimethyi-3-( 1OH-phenothiazin-10-yl)propan- alimemazine and im purity C.
1-am ine hemi [(2Æ,3iQ-2,3-dihydroxybutanedioate]. Calculation of percentage contents:
C o n te n t — correction factors: multiply the peak areas o f the following
99.0 per cent to 101.0 per cent (dried substance). impurities by the corresponding correction facto r
impurity A = 4.4; impurity C = 0.4;
CHARACTERS — for each impurity, use the concentration o f alimemazine
A p p e a ra n c e in reference solution (a).
W hite or very slightly yellowish powder. Limits:
S o lu b ility — impurity B: máximum 0.3 per cent;
F red y soluble in water, sparingly soluble in ethanol — impurities A, C: for each impurity, maximum
(96 per cent), practically insoluble in toluene. 0.15 p er cent;
It deteriorates when exposed to air and light. — unspecified impurities: for each impurity, maximum
0.10 p er cent;
ID E N T IF IC A T IO N
— total: maximum 0.5 p er cent;
Infrared absorption spectrophotom etry (2.2.24). — reporting threshold: 0.05 per cent.
Comparison alimemazine hemitartrate CRS. Loss on drying (2.2.32)
TESTS M axim um 0.5 per cent, determined on 1.000 g by drying in
A p p e a ra n c e o f so lu tio n an oven at 105 °C for 3 h.
T he solution is not m ore opalescent than reference Sulfated ash (2.4.14)
suspension II (2.2.1) and n o t more intensely coloured than M axim um 0.1 per cent, determ ined on 1.0 g.
reference solution BY5 (2.2.2, Method IT).
2016 Allantoin 1-95
enantiomer TESTS
3 and
S o lu tio n S
Dissolve 0.5 g in carbon dioxide-free water R, with heating if
necessary, and dilute to 100 m L with the same solvent.
A cid ity o r a lk a lin ity
B . (2i?S}-ATj2-dimethyl-3-( 1OH-phenothiazin-10-yl)propan-1-
T o 5 m L o f solution S add 5 m L of carbon dioxide-free
amine,
water R, 0.1 m L o f methyl red solution R and 0.2 m L of
0.01 M sodium hydroxide. T he solution is yellow. Add 0.4 m L
o f 0.01 M hydrochloric acid The solution is red.
O p tic a l ro ta tio n (2.2.7)
T h e angle of optical rotation, determined on solution S, is
-0 .1 0 ° to + 0.10°.
R ed u c in g su b sta n c e s
Shake 1.0 g with 10 m L o f water R for 2 min. Filter.
C. 1OH-phenothiazine. Add 1.5 m L o f 0.02 Mpotassium permanganate. T he solution
Ph Bur m ust rem ain violet for at least 10 min.
R e la te d su b sta n c e s
Examine by thin-layer chromatography (2.2.27), using a
suitable cellulose for chromatography R as the coating
★ ★ substance.
Allantoin ★ ★
Test solution (a) Dissolve 0.10 g o f the substance to be
(Ph. Eur. monograph 1288) * * * * * examined in 5.0 m L of water R with heating. Allow to cool.
Dilute to 10 m l, with methanol R. Use the solution immediately
o after preparation.
f H-
H. . y ' NH and enantiomer Test solution (b) Dilute 1 m L of test solution (a) to 10 m L
H,N with a mixture o f 1 volume of methanol R and 1 volume o f
water R.
Reference solution (a) Dissolve 10 m g of allantoin CRS in a
€4 ^ 403 158.1 97-59-6 mixture o f 1 volume o f methanol R and 1 volume of water R
and dilute to 10 m L with the same mixture of solvents.
A ctio n a n d u se
Astringent; keratolytic. Reference solution (b) Dissolve 10 m g of urea R in 10 m L o f
water R. Dilute 1 m L o f this solution to 10 m L with
PhEir ___________________ methanol R
D E F IN IT IO N Reference solution (c) M ix 1 m L o f reference solution (a) and
Allantoin contains n o t less than 98.5 per cent and not more 1 m L of reference solution (b).
than the equivalent o f 101.0 per cent of Apply to the plate 10 jiL o f test solution (a) and 5 |iL each
(RS) -(2 j 5-dioxoimidazolidin-4-yl)urea. o f test solution (b), reference solution (a), reference
solution (b) and reference solution (c). Develop over a path
CHARACTERS
o f 10 cm using a mixture of 15 volumes of glacial acetic
A white or alm ost white, crystalline powder, slightly soluble
add R, 25 volumes o f water R and 60 volumes of butanol R.
in water, very slightly soluble in alcohol.
1-96 Allergen products 2016
Allow the plate to dry in air. Spray the plate with a 5 g/L prepared by dilution of aqueous or glycerinated extracts, or
solution of dimethylaminobenzaldehyde R in a mixture of by reconstitution o f unmodified freeze-dried extracts.
1 volume of hydrochloric add R and 3 volumes of methanol R. F or specific immunotherapy, allergen products may be either
Dry the plate in a current o f h o t air. Examine in daylight unmodified extracts or extracts modified chemically and/or
after 30 min. Any spot in the chromatogram obtained with by adsorption onto different carriers (for example, aluminium
test solution (a), apart from th e principal spot, is not more hydroxide, calcium phosphate or tyrosine).
intense than the spot in the chrom atogram obtained with
reference solution (b) (0.5 p er cent). T h e test is not valid P R O D U C T IO N
unless the chrom atogram obtained with reference solution (c) S O U R C E M A T E R IA L S
shows two clearly separated principal spots. Source materials for the preparation of allergen products are
products o f animal o r vegetable origin, mostly pollens,
L oss o n d ry in g ( 2.2.32)
moulds, mites, animal epithelia and outgrowths (such as hair
N ot more than 0.1 p e r cent, determined on 1.000 g by
and feathers) and/or dander, hymenoptera venoms, insects
drying in an oven at 105 °C.
and certain foods.
S u lfa te d a s h (2.4.14)
W here allergen products are m anufactured using materials of
N ot more than 0.1 p e r cent, determined on 1.0 g.
hum an or animal origin, the requirements o f chapter 5.1.7.
ASSAY Viral safety apply.
Dissolve 120.0 mg in 40 m L o f water R. T itrate with 0.1 M T h e source materials are defined by their origin, nature,
sodium hydroxide, determining the end-point m ethod o f collection or production and pretreatm en t
potentiometrically (2.2.20). Control m ethods and acceptance criteria relating to identity
1 m L of 0.1 M sodium hydroxide is equivalent to 15.81 m g of and purity are established. T h e acceptance criteria m ust
C 4H 6N 4O 3. ensure the consistency o f the allergenic source material from
a qualitative and quantitative point o f view. T h e source
IM P U R IT IE S
materials are stored under controlled conditions justified by
H CO2H stability data.
To T h e collection or production, as well as the handling o f the
source materials are such that uniform composition is
A. glyoxylic acid, ensured as far as possible from batch to batch.
T h e content of the relevant residual solvents, heavy metals
o and pesticides is determ ined on a num ber o f batches
according to a justified sampling plan. Residual solvents and
pesticides are limited according to the principles defined in
general chapter 2.4.24. Identification and control of residual
B. carbamide (urea).
solvents and 2.8.13. Pesticide residues respectively.
--------------------------------- — .— _____________________________PhEur
P o lle n s
Potential chemical contam inants, such as pesticides, heavy
metals and solvents, m ust be minimised. T h e content of
foreign pollen m ust be limited to 1 per cent o f total mixed
Allergen Products ***** pollens and 0.5 per cent o f any individual pollen as
determined by a microscopic particle count. D etectable
(Ph Eur. monograph 1063) * m ould spores m ust not exceed 1 per c e n t
Ph Fur _________________________________________ T h e contam ination with particles o f plant origin other than
This monograph does not apply to: chemicals that are used solely pollen m ust be kept to a minim um. T he m axim u m allowed
for diagnosis of contact dermatitis; chemically synthesised products; contam ination m ust be justified.
allergens derived by recombinant DNA technology. It does not M o u ld s
necessarily apply to allergen products for veterinary use. Biologically active contam inants such as mycotoxins in
D E F IN IT IO N moulds m ust be m in im ised and any presence justified.
Appropriate measures have to be im plem ented to avoid
Allergen products are pharm aceutical preparations derived
c o n tam inatio n by foreign m ould strains. C are m ust be taken
from extracts of naturally occurring source materials
to minimise any allergenic constituents o f the media vised for
containing allergens, which are substances that lead to and/or
provoke allergic reactions. T h e allergenic components are the cultivation of moulds as source materials. Culture media
th at contain substances of hum an or anim al origin m ust be
most often o f a proteinaceous nature. Allergen products are
intended for in vivo diagnosis o r treatm ent o f allergic diseases justified and, when required, m ust be suitably treated to
attributed to these allergens. ensure the inactivation or elimination of possible
transmissible agents of disease.
Allergen products are available as finished products, and as
finished products used on a nam ed-patient basis. Allergen T h e production m ethod is validated to demonstrate that
products are generally presented as parenteral preparations, allergen products obtained from moulds and intended for
parenteral administration, if tested* would comply with the
eye preparations, preparations for inhalation, preparations for
test for abnormal toxicity for immunosera and vaccines for
oral use, sublingual preparations or preparations for skin
tests. hum an use (2.6.9).
th at contain substances of hum an or animal origin m ust be of suitable reagents, as well as the intended use. The characterised
justified and, when required, must be suitably treated to IHRP is used as the reference in the batch control of active
ensure the inactivation or elimination of possible substances and intermediates and, if possible, in the batch control
transmissible agents of disease. offinished products.
Anim al epithelia and outgrowths and/or dander T he IH R P is characterised by the protein content
T hey are obtained from healthy animals selected to avoid determination and a protein profile using appropriate
possible transmissible agents of disease. methods (such as isoelectric focusing, sodium dodecyl sulfate
Hymenoptera venoms polyacrylamide gel electrophoresis, Immunoelectrophoresis,
T h e species of hymenoptera from which the venom is capillary electrophoresis, chromatographic techniques and
mass spectrometry).
extracted is identified and specified. T h e m ethods of insect
collection and venom extraction are described and m ust Allergenic components may be detected by appropriate
ensure that the source material is of proper quality. methods (for example, immunoblotting or crossed radio-
im munoelecupphoresis). Characterisation of the allergenic
Food
components may include identification o f relevant allergens
T h e scientific nam e (species, variety, strain etc.) o f the
based on serological or other techniques using pooled or
an im al or vegetable species is indicated and the p art used is
individual sera from allergic patients, or allergen-specific
stated, if applicable. Foods must be o f a quality suitable for
polyclonal or monoclonal antibodies.
hum an consum ption. T he origin o f the food stuff as well as
its processing stage is stated. Determ ination o f the content of relevant allergens is
performed wherever possible. T his determination may be
MANUFACTURING PROCESS m ade using individual allergen-specific reference standards,
Allergen products are generally obtained by extraction, and w hen available. T h e choice of the relevant allergen
m ay be purified, from the source materials using appropriate components subjected to the determination m ust be justified.
m ethods shown to preserve the allergenic properties o f the Individual allergens are identified and named according to
com ponents. Allergens for which there are not enough internationally established nomenclature wherever possible.
patients to determine the total allergenic activity in vivo or in
T h e biological potency o f the first IH R P is determined in
vitro, the extraction ratio indicating the relative proportions
patients by in vivo techniques such as skin testing, and
(mlV) o f allergenic source materials and solvents is a
expressed in units of biological activity except when not
minimum requirem ent. Allergen products presented as
enough patients are available. In this case, the potency of the
parenteral preparations, eye preparations, preparations for
first IH R P is determ ined by an m vitro method.
inhalation and preparations for skin testing are m anufactured
Subsequently, the biological activity of future EHRPs is
u nder aseptic conditions.
demonstrated by m vitro methods by comparison with the
In the manufacture, packaging, storage and distribution o f results obtained with the first IH R P. T he in vitro potency
allergen products intended for administration by other routes, may be measured by a suitable immunoassay (for example,
suitable measures are taken to ensure their microbial quality; an assay based on the inhibition o f the binding capacity of
recom m endations on this aspect are provided in chapter specific immunoglobulin E antibodies).
5.1.4. Microbial quality of non-sterUe pharmaceutical preparations
and substances for pharmaceutical use. IDENTIFICATION
T h e tests for identification are performed as late as possible
All allergen preparations are m anufactured under conditions
in the manufacturing process. In the case of products used
designed to minimise exogenous and endogenous enzymatic
on a nam ed-patient basis, the control is performed on the
degradation.
active substance and/or at the intermediate stage between the
Any purification procedure is designed to minimise the active substance and the finished product.
content o f any potential irritant low molecular mass
Identity is confirmed by comparison with the IH R P using
com ponents and non-allergenic components.
protein profiling by appropriate methods (for example,
Allergen products m ay contain suitable antimicrobial isoelectric focusing, sodium dodecyl sulfate polyacrylamide
preservatives, the nature and concentration of which have to gel electrophoresis, Immunoelectrophoresis, immunoblotting,
be justified. liquid chromatography o r mass spectrometry).
T h e manufacturing process comprises various stages: In exceptional cases, if no IH R P is available, a representative
— source material; batch may be used to confirm identity.
— active substance: it is generally a modified or an
Identity may also be confirmed by comparison with
unm odified allergen extract; where applicable it is stored
individual allergen-specific reference standards, when
under conditions ensuring its stability, for example freeze-
available.
dried;
— finished product. TESTS
All other stages of the manufacturing process are considered T h e tests are perform ed as late as possible in the
as intermediates. manufacturing process. In the case o f products used on a
named-patient basis, th e control is performed on the active
IN-HOUSE REFERENCE PREPARATION substance and/or at the intermediate stage between the active
An appropriate representative preparation is selected as the substance and the finished product.
in-house reference preparation (IHRP), characterised and
Various biochemical and immunological tests have been
used to verify batch-to-batch consistency. T he IH R P is
developed in order to characterise allergens qualitatively and
stored in suitably sized aliquots under conditions ensuring its
quantitatively. In those cases where such m ethods cannot be
stability, for example freeze-dried.
applied, particularly for the determination o f allergenic
Characterisation o f the in-house reference preparation activity and allergen and/or protein profile, justification m ust
The extent of characterisation of the IHRP depends on the source be provided.
material, knowledge of the aUergejric components and availability
1-98 Allopurinol 2016
of salicyUc add in place o f the solution being examined and Reference solution (b) Dissolve 10 m g o f alprazolam CRS and
b eg in n in g at the words ‘add 4 m L o f ừon(w) chloride 10 m g o f midazolam CRS in methanol R and dilute to 10 m L
solution..’. Each g o f salicylic a d d is equivalent to 1.305 g of with the same solvent.
C 9Ỉ Ỉ 8O 4. Plate TLC silica gel GF2 5 4 plate R.
Mobile phase glacial acetic acid R, water R, methanol R, ethyl
acetate R (2:15:20:80 VIV/VIV).
Application 5 |iL.
Alprazolam ★ ★ Development Over a path o f 12 cm.
★ ★
(Ph. Eur. monograph 1065) ***** Drying In. air.
Detection Examine in ultraviolet light at 254 nm.
H3C ^ w System suitability: reference solution (b):
— the chromatogram shows 2 clearly separately spots.
Results T h e principal spot in the chrom atogram obtained with
the test solution is similar in position and size to the principa]
spot in the chromatogram obtained with reference
solution (a).
TESTS
Related substances
Liquid chromatography (2.2.29).
C17H13aN4 308.8 28981-97-7
Buffer solution Dissolve 7.7 g of ammonium acetate R in
1000 m L o f water R and adjust to p H 4.2 with glacial acetic
Action and use
Benzodiazepine. add R.
Test solution Dissolve 0.100 g o f the substance to be
PhEir __________________________________________________________ examined in dimethylformamide R and dilute to 10.0 m L with
DEFINITION the same solvent.
8-Chloro-1-m ethyl-6-phenyl-4/i- [ 1,2,4] triazolo [4,3-a] Reference solution (a) Dissolve 2 m g o f alprazolam CRS and
[1,4] benzodiazepine. 2 m g o f triazolam CRS in dimethylformamide R and dilute to
100.0 m L with the same solvent.
Content
99.0 per cent to 101.0 per cent (dried substance). Reference solution (b) Dilute 5.0 m L o f the test solution to
100.0 m l. with dimethylformamide R. Dilute 0.5 m L o f this
CHARACTERS solution to 10.0 m L with dimethylformamide R.
Appearance
Cdumn:
W hite or almost white, crystallin e powder.
— size. I = 0.25 m, 0 = 4.6 mm;
Solubility — stationary phase: phenylsüyl silica gelfor chromatography R1
Practically insoluble in water, freely soluble in methylene (5 nm).
chloride, sparingly soluble in acetone and in ethanol Mobile phase:
(96 per cent). — mobile phase A: buffer solution, methanol R (44:56 VIV)\
It shows polymorphism (5.9). — mobile phase B: buffer solution, methanol R (5:95 V!V)\
IDENTIFICATION — temperature:. 40 °C;
First identification B.
Second identification A , C. lim e Mobile phaae A Mobile phase B
(min) (percent V/V) (ptT cent V7V)
A. Dissolve the substance to be examined in the smallest
0 -1 5 98 2
necessary quantity o f ethyl acetate R and evaporate to dryness
on a water-bath. Thoroughly mix 5.0 m g o f the substance to 15 -3 5 98 -> 1 2 + 99
be examined with 5.0 mg of alprazolam CRS. T he melting 3 5 -4 0 1 99
point (2.2.14) o f the mixture does n o t differ by more than
2 °C from the melting point o f the substance to be
examined. Flow rate 2 mL/min.
B. Infrared absorption spectrophotometry (2.2.24). Detection Spectrophotom eter at 254 nm.
Preparation Discs. Irgecdon 10 |iL; inject dtmethylformamide R a sa blank.
Comparison alprazolam CRS. Retention time Triazolam = about 9 min;
If the spectra obtained in the solid state show differences, alprazolam = about 10 min.
dissolve the substance to be examined and the reference System suitability: reference solution (a):
substance separately in the m in im u m volume of ethyl — resolution: minim um 1.5 between the peaks due to
acetate R, evaporate to dryness on a water-bath and record triazolam and alprazolam.
new spectra using the residues. Limits:
C. Thin-layer chromatography (2.2.27). — totah not more than the area o f the principal peak in the
Test solution Dissolve 10 m g o f the substance to be exam in ed chrom atogram obtained with reference solution (b)
in methanol R and dilute to 10 m L w ith the same solvent. (0.25 per cent);
— disregard lima: 0.2 times the area o f the principal peak in
Reference solution (a) Dissolve 10 m g o f alprazolam CRS in
the chromatogram obtained with reference solution (b)
methanol R and dilute to 10 m L with the same solvent.
(0.05 per cent).
2016 Alprazolam 1-105
L oss o n d ry in g (2.2.32)
M axim um 0.5 per cent, determined on 1.000 g by drying in
an oven at 105 °C.
S u lfa te d a s h (2.4.14)
Maximum 0.1 per cent, determined on 1.0 g.
A SSA Y
Dissolve 0.140 g in 50 m L of a mixture o f 2 volumes of
acetic anhydride R and 3 volumes of anhydrous acetic acid R.
Titrate with 0.1 M perchloric acid, determining the end-point G. 7-chloro- l-methyl-5-phenyl[ 1,2,4]triazolo [4,3-a] quinolin-
potentiometrically (2.2.20). Titrate to the 2nd point of 4-amine,
inflexion.
1 m L of 0.1 M perchloric add is equivalent to 15.44 mg
o f C 17H 13CIN 4.
STO R A G E
Protected from light.
IM P U R IT IE S
N y CH3
H . bis[[4-(2-ben2oyl-4-chlorophenyl)-5-methyl-4H'-lj2}4-
and enantiomer triazol-3-yl] methyl] amine.
OH
A. (4R5)-3-amino-6-chloro-2-methyl-4-phenyl-3,4-
dihydroquinazolin-4-olj
and enantiomer
E. (2 -amino-5-chlorophenyl)phenylmethanone,
1-106 Alprenolol Hydrochloride 2016
C. Examine the chrom atograms obtained in the test for the mobile phase.
im purity D. Reference solution (a) Dissolve 4.0 m g o f alprenolol
Detection Examine in daylight, after exposure to iodine hydrochloride CRS and 0.8 m g of 4-isopropylphenol R in the
vapour for 30 min. mobile phase and dilute to 100.0 m L with the mobile phase.
Results T he principal spot in the chrom atogram obtained with Reference solution (b) D ilute 4.0 m L of the test solution to
100.0 m L with die mobile phase. D ilute 1.0 m L o f this
test solution (b) is similar in position, colour and size to the
principal spot in the chrom atogram obtained with reference solution to 10.0 m L with the mobile phase.
solution (a). Column:
— size. I = 0.15 m , 0 = 4 mm;
D . It gives reaction (a) of chlorides (2.3.1).
— stationary phase, octylsüyl silica gel for chromatography R
TESTS (5 pm).
Solution S Mobile phase M ix 0.656 g o f sodium octanesidfonate R with
Dissolve 1.0 g in carbon dioxide-free water R and dilute to 150 mT. o f acetomtrüe R and dilute to 500 m L with
50 m L with the same solvent. phosphate buffer p H 2.8 prepared as follows: mix 1.78 g of
Appearance of solution phosphoric add R and 15.6 g o f sodium dihydrogen phosphate R
Solution S is clear (2.2.1) and n o t m ore intensely coloured and dilute to 2000 m L w ith water R
than reference solution B9 (2.2.2, Method II). Flow rate 1 mlVmin.
Acidity or alkalinity Detection Spectrophotom eter at 280 nm .
T o 10 m L o f solution S add 0.2 mT. o f methyl red solution R Equilibration W ith the mobile phase for about 1 h.
and 0.2 m L o f 0.01 M hydrochloric acid; the solution is red.
Injection 20 nL.
A dd 0.4 m L o f 0.01 M sodium hydroxide; the solution is
yellow. Run time Twice the retention time of alprenolol.
Impurity C Retention time Alprenolol = about 11 min;
M axim um 0.1 per c e n t 4-isopropylphenol = about 18 min.
Reference solution (b) Dissolve 1.0 m g o f dinoprostone Time Mobile phase A Mobile phase B
impurity C CRS (alprostadil impurity H ) and 1.0 m g o f the (min)____________ (percent V7V)________ (per cent V7V)
substance to be examined in a mixture o f equal volumes of 0 -5 0 100 0
acetomtrile R1 and water R and dilute to 20.0 m L with the 50-51 100 + 0 0+100
sam e mixture o f solvents.
51 -61 0 100
Reference solution (c) In order to prepare impurities A and B
in situ, dissolve 1 mg o f the substance to be examined in 61 -62 0+100 100 + 0
100 pL o f 1 M sodium hydroxide (the solution becomes 6 2 -7 2 100 0
brownish-red), wait for 3 min and add 100 |iL o f a 112 g/L
solution o f phosphoric add R (yellowish-white opalescent
solution); dilute to 5.0 m L with a mixture of equal volumes Relative retention W ith reference to alprostadil (retention
o f acetomtrUe R1 and water R. time = about 7 min): impurity A = about 2.4;
S y ste m A impurity B = about 2.6.
Column: System suitability:
— size: I = 0.25 m , 0 = 4.0 mm; — resolution: m in im u m 1 .5 between the peaks due to
— stationary phase: base-deactivated octylsUyl silica gel for impurity A and impurity B in the chrom atogram obtained
chromatography R (4 pm) with a pore size of 6 nm ; w ith reference solution (c).
— temperature: 35 °C. C arry out the test according to system A and B.
Mobile phase: Limits:
— mobile phase A: dissolve 3.9 g of sodium dihydrogen — correction factors: for the calculation of content, multiply
phosphate R in water R and dilute to 1.0 L with the same the peak areas o f the impurities listed in Table 1488.-1 by
solvent; adjust to p H 2.5 with a 2.9 g/L solution of the corresponding correction factor;
phosphoric add R (approximately 600 m L is required);
to 740 m L o f the buffer solution add 260 m L o f Table 1488.-1.
acetomtrUe R1;
Imparity Relative retention Relative retention Correction factor
— mobile phase B: dissolve 3.9 g o f sodium dihydrogen (system A) (system B)
phosphate R in water R and dilute to 1.0 L with the same impurity G 0.80 - 0.7
solvent; adjust to p H 2.5 with a 2.9 g/L solution o f
impurity F 0.88 - 0.8
phosphoric add R (approximately 600 m L is required);
to 200 m L o f the buffer solution add 800 m L of impurity D 0.90 - 1.0
acetomtrUe R1; impurity H 0.96 - 0.7
Flow rate 1 mL/m in. — impurity A: n o t more than 3 times the area of the
principal peak in the chrom atogram obtained with
Detection Spectrophotom eter at 200 nm .
reference solution (a) (1.5 per cent);
Injection 20 |iL. — impurity B: n o t more than the area o f the principal peak
Retention time Alprostadil = about 63 m in . in the chrom atogram obtained with reference solution (a)
System suitability: (0.5 per cent);
— resolution: m in im u m 1.5 between the peaks due to — any other impurity: n o t m ore than 1.8 times the area o f the
im purity H and alprostadil in the chromatogram obtained principal peak in the chrom atogram obtained with
with reference solution (b). reference solution (a) (0.9 per cent), and n o t more than 1
S y ste m B such peak has an area greater than the area o f the
U se the same conditions as for system A with the following principal peak in the chrom atogram obtained with
mobile phase and elution programme: reference solution (a) (0.5 per cent). Evaluate impurities
— mobile phase A: dissolve 3.9 g of sodium dihydrogen appearing at relative retentions less than 1.2 by system A
phosphate R in water R and dilute to 1.0 L with the same and impurities appearing at relative retentions greater
solvent; adjust to p H 2.5 with a 2.9 g/L solution o f than 1.2 by system B;
phosphoric add R (approximately 600 m L is required); — total: not more than 3 times the area o f the principal peak
to 600 m L o f the buffer solution add 400 m L of in the chromatogram obtained with reference solution (a)
acetomtrUe R l; (1.5 per cent);
— mobile phase B: use mobile phase B as described under — disregard limit. 0.1 times the area of the principal peak in
system A; the chromatogram obtained with reference solution (a)
(0.05 per cent).
W a te r ( 2.5.32)
M axim um 0.5 per cent, determ ined on 50 mg.
2016 Alprostadil 1-109
ASSAY
Liquid chromatography (2.2.29) as described in the test for
related substances, system A. Prepare the solutions protected
from Ught.
Test solution Dissolve 10.0 m g of the substance to be H OH
examined in a mixture of equal volumes of acetonitrile R1 and
water R and dilute to 25.0 m L with the same mixture of E. 7-[(li?,2J?,35)-3-hydroxy-2-[(l£,35)-3-hydroxyoct-l-
solvents. Dilute 3.0 m L of the solution to 20.0 m L with a enyl]-5-oxocyclopentyl]heptanoic acid
mixture o f equal volumes of acetonitrile R1 and water R. (11-epiprostaglandin E i),
Reference solution Dissolve 5.0 mg of alprostadil CRS in a
mixture of equal volumes of acetonitrile R1 and water R and
dilute to 25.0 m L with the same mixture o f solvents. Dilute
6.0 m L o f the solution to 20.0 m L with a mixture of
equal volumes o f acetonitrile R1 and water R.
HO"
Injection 20 ^L. H OH
Calculate the percentage content of C 20H 34O 5 taking into
account the assigned content of alprostadil CRS.
F . 7-[(l S,2R,3R)-3-hydroxy-2-[(1 £ ,3 6 )-3-hydroxyoct-1-
ST O R A G E enyI]-5-oxocyclopentyl]heptanoic a d d
At a tem perature o f 2 °C to 8 °C. ( 8-epiprostaglandin E i),
IM P U R IT IE S
H ’oh
H OH
G . (5Z)-7- [(1 i?,2i?3^)-3-hydroxy-2-[(l£,35)-3-hydroxyoct-
A 7- [(1 R,2S)-2- [(1 £,3.S)-3-hydroxyoct-l -enyI]-5- l-enyI]-5-oxocydopentyl]hept-5-enoic a d d
oxocyclopent-3-enyI]heptanoic acid (prostaglandin A 0 , (dinoprostone),
O
CO2H
CH3
H OH H OH
H OH
C. 7-[(li?,2i?,3i?)-3-hydroxy-2-[(l£)-3-oxooct-l-enyI]-5-
oxocydopentyl]heptanoic a d d (15-oxoprostaglandin E j), I. ethyl 7-[(1 i?,2i?,3i?)-3-hydroxy-2- [(1 £ ,3 5 )-3-hydroxyoct-
l-enyI]-5-oxocydopentyi]heptanoate (prostaglandin E i,
ethyl ester),
H OH
D . 7-[(li?,2i?,3i?)-3-hydroxy-2-[(l£*3.R)-3-hydroxyoct-l- H OH
enyl]-5-oxocyclopentyl]heptanoic a d d
(15-epiprostagJandin E i),
J. 1-methylethyl 7-[(li?,2i?,3i?)-3-hydroxy-2-[(l£,36)-3-
hydroxyoct-l-enyl]-5-oxocyclopentyl]heptanoate
(prostaglandin E i, isopropyl ester),
1-110 Alteplase for Injection 2016
dimensional separation of glycopeptide variants, lot-to-lot and reference standard and using a relative molecular mass of
consistency of the microheterogeneity o f glycosylation can be 180.2 for mannose and a relative molecular mass of 59 050
demonstrated. for the alteplase protein moiety. T h e neutral sugar content of
T he tryptic-peptide map o f alteplase samples must be the alteplase samples m ust be in the range o f 70 to
consistent with the tryptic-peptide map of alteplase reference 130 per cent compared to alteplase reference standard, which
standard. contains about 12 moles of neutral sugar per mole of
alteplase.
Monomer content
T h e m onom er content of alteplase is m easured by gel- CHARACTERS
perm eation liquid chromatography under non-reduced W hite or slightly yellow powder or solid friable mass.
conditions as described under M onomer content (see Tests). Reconstitute the preparation as stated on the label immediately
T he m onom er content of alteplase bulk samples must be before carrying out the Identification, Tests (except those for
higher than 95 per cent solubility and water) and Assay.
Type I/Type II alteplase content IDENTIFICATION
C H O cells produce 2 glycosylation variants o f alteplase. A. T he assay serves also to identify the preparation.
Type I alteplase contains 1 polymannose-type glycosylation at
B. Tryptic-peptide mapping. Examine by liquid
position Asn 117 and 2 complex-type glycosylation sites at
chromatography (2.2.29).
positions Asn 184 and Asn 448. Type II alteplase is only
glycosylated at positions Asn 117 and Asn 448. Test solution Dilute the preparation to be examined with
water R to obtain a solution containing about 1 mg of
T he ratio o f Type I/Type II alteplase is constant in the range
alteplase per millilitre. Dialyse about 2.5 m L o f the solution
o f 45 to 65 per cent of Type I and 35 to 55 per cent of
for at least 12 h into a solution containing 480 g/L of urea R,
Type II. T h e content of alteplase Type I and Type II can be
44 g/L o f tris(hydroxymethyl) aminomethane R and 1.5 g/L of
determ ined by a densitometric scan o f SDS-PAGE (sodium
sodium edetate R and adjusted to p H 8.6, using a mem brane
dodecyl sulfate polyacrylamide gel electrophoresis) gel.
with a cut-oflf point corresponding to a relative molecular
Plasmin-treated samples of alteplase, which are reduced and
mass o f 10 000 for globular proteins. Measure the volume of
carboxymethylated before loading on the gel, are separated
the solution, transfer it to a clean test-tube and add per
into 3 bands: Type I alteplase A-chain (AA 1-275), Type II
millilitre 10 |iL o f a 156 g/L solution of dithiothreitol R. Allow
alteplase A-chain (AA 1-275) and alteplase B-chain
to stand for 4 h, cool in iced water and add per millilitre of
(AA 276-527). T h e ratio o f Type I/Type II alteplase is
solution 25 pL o f a freshly prepared 190 g/L solution of
determ ined from a calibration curve, which is obtained by a
iodoacetic add R. Allow to stand in the dark for 30 min.
densitometric scan of defined mixtures o f purified Type I
A dd per millilitre 50 |oL of dithiothreitol solution to stop the
alteplase and T ype II alteplase standards.
reaction. Dialyse for 24 h against an 8 g/L solution of
SDS-PAGE ammonium hydrogen carbonate R. Add 1 part of trypsin for
SD S-PAG E (silver staining) is used to demonstrate purity of peptide mapping R to 100 parts o f the protein and allow to
the alteplase bulk material and the integrity o f the alteplase stand for 6 h to 8 h. Repeat the addition of trypsin and allow
molecule. F or alteplase bulk samples, no additional protein to stand for a total of 24 h.
bands compared to reference standard or degradation
Reference solution Prepare as for the test solution using a
products m ust occur in SDS-PAGE gels at a loading am ount
suitable reference standard instead of the preparation to be
o f 2.5 |xg alteplase protein per lane and a limit of detection of
examined.
5 ng per protein (BSA) band.
T he chromatographic procedure may be carried out using:
Bacterial endotoxins (2.6.14) — a column 0.1 m long and 4.6 mm in internal diameter
Less than 1 IU p er milligram of alteplase. packed with octadecylsüyl silica gelfor chromatography R
Sialic adds (5 jim to 10 pm);
Proceed using a suitable validated m ethod devdoped Mobile phase A 8 g/L solution of sodium dikydrogen
according to general chapter 2.2.59. Glycan analysis of phosphate R , adjusted to p H 2.85 with phosphoric
glycoproteins. T h e sialic ad d s content for the test samples acid R, filtered and degassed;
m ust be in the range of 70 to 130 per cent compared to Mobile phase B 75 per cent V/V solution of
alteplase reference standard, which contains about 3 moles of acetomtrUe R in mobile phase A;
sialic a d d s per mole of alteplase. — as detector a spectrophotometer set at 210 nm.
Neutral sugars Equilibrate the system with mobile phase A at a flow rate of
Dilute alteplase samples and the reference standard in the 1 mL/min. After injection of the solution, increase the
assay buffer, containing 34.8 g/L of arginine R, 0.1 g/L of proportion of mobile phase B at a rate of 0.44 per cent per
polysorbate 80 R and adjusted to pH 7.4 with phosphoric m inute until the ratio of mobile phase A to mobile phase B is
acid R, to a protein concentration of 50 [ig/m L Prepare the 60:40, then increase the proportion of mobile phase B at a
following concentrations o f mannose in the same assay buffer rate o f 1.33 per cent per minute until the ratio o f mobile
for a calibration curve: 20, 30, 40, 50 and 60 (ig/mL. Pipette phase A to mobile phase B is 20:80 and then continue
2 m L of alteplase samples and reference standard, as well as elution with this mixture for a further 10 min. Record the
2 m L o f each mannose concentration in duplicate in reagent chromatogram for the reference solution: the test is not valid
tubes. A dd 50 jaL of phenol R, followed by 5 m L of sulfuric unless the resolution of peaks 6 (peptides 268-275) and 7
acid R, in each reagent tube. Incubate the mixture for 30 m in (peptides 1-7) is at least 1.5; whf and are n o t more than
at room tem perature. M easure the absorbance at 492 nm for 0.4 min. Inject about 100 |iL o f the test solution and record
each tube. Read the content of neutral sugars from the the chromatogram. Verify the identity of the peaks by
mannose calibration curve. T he neutral sugar content is comparison with the chromatograms of the reference
expressed in moles of neutral sugar per mole of alteplase, solution. There should n o t be any additional significant peaks
taking into account the dilution factor for alteplase samples o r shoulders, a significant peak or shoulder being defined as
1-112 Alteplase for Injection 2016
one with an area response equal to or greater than 5 p er cent — a colum n 0.6 m long and 7.5 m m in internal diam eter
of peak 19 (peptides 278-296); no significant peak is missing. packed with silica-based, rigid, hydrophilic gel with
A type chromatogram for identification of the peaks cited is spherical particles 10 pm to 13 pm in diam eter, suitable
shown in Figure 1170.-1. for size-exclusion chromatography;
— as mobile phase at a flow rate o f 0.5 m L/m in a solution
TESTS
c o n tainin g 30 g/L of sodium dihydrogen phosphate R and
A p p e a ra n c e o f so lu tio n
1 g/L o f sodium dodecyl sulfate R, adjusted to p H 6.8 with
T he reconstituted preparation is clear (2.2. I) and n o t more
dilute sodium hydroxide solution R;
intensely coloured than reference solution Y7 (2.2.2,
— as detector a spectrophotom eter set at 214 nm .
Method II).
Inject about 50 pL o f the test solution and record the
p H (2.2.3)
chrom atogram. T he chrom atogram shows 2 m ajor peaks
7.1 to 7.5. corresponding to single-chain and two-chain alteplase.
S o lu b ility Calculate the relative am ount o f single-chain alteplase from
A dd the volume o f the liquid stated on the label. the peak area values.
T h e preparation dissolves completely within 2 m in at 20 °C T h e test is n o t valid unless: the num ber o f theoretical plates
to 25 °C. calculated on the basis o f the single-chain alteplase peak is at
P r o te in c o n te n t least 1000. T h e content of single-chain alteplase is n o t less
Prepare a solution of the substance to be examined with an than 60 per cent o f the total am ount o f alteplase-related
accurately known concentration of about 1 g/L. Using a substances found.
34.8 g/L solution of arginine R adjusted to p H 7.3 with M o n o m e r c o n te n t
phosphoric acid R, dilute an accurately measured volume o f Examine by liquid chromatography (2.2.29).
the solution o f the substance to be examined so that the
Test solution Reconstitute the preparation to be examined to
absorbance measured at the maximum at about 280 nm is
obtain a solution containing about 1 m g per millilitre.
0.5 to 1.0 (test solution). M easure the absorbance (2.2.25) o f
the solution at the maximum at about 280 nm and at T h e chromatographic procedure may be carried out using:
320 nm using the arginine solution as the compensation — a colum n 0.6 m long and 7.5 m m in internal diam eter
liquid. Calculate the protein content in the portion o f packed with silica-based rigid, hydrophilic gel with
alteplase taken from the following expression: spherical particles 10 pm to 13 pm in diameter, suitable
for size-exclusion chromatography;
— as mobile phase at a flow rate o f 0.5 m L/m in a solution
V (-^280 ~ -<4.320)
containing 30 g/L of sodium dihydrogen phosphate R and
1.9
1 g/L o f sodium dodecyl sulfate R, adjusted to p H 6.8 with
dilute sodium hydroxide solution R;
in which V is the volume o f th e test solution, A 2 S0 is the
— as detector a spectrophotom eter set at 214 nm .
absorbance at the maximum at about 280 nm and A 32q is the
absorbance at 320 nm. Inject the test solution and record the chromatogram.
T h e test is n o t valid unless the num ber o f theoretical plates
Single-chain c o n te n t
calculated for the alteplase m onom er peak is at least 1000.
Examine by liquid chrom atography (2.2.29).
M easure the response for all peaks, i.e. peaks corresponding
Test solution Dissolve the preparation to be examined in to alteplase species o f different molecular masses. Calculate
water R to obtain a solution containing about 1 m g o f the relative content of m onom er from the area values of these
alteplase per millilitre. Place about 1 m L o f the solution in a peaks. T h e m onom er content for alteplase m ust be at least
tube, add 3 m L of a 3 g/L solution o f dxthiotkreitd R in the 95 p er c e n t
mobile phase, place a cap on the tube and heat at about
80 °C for 3 m in to 5 min.
T h e chromatographic procedure may be carried out using:
2016 Altizide 1-113
PhEur__________________________________________________________ PhEtr.
D E F IN IT IO N D E F IN IT IO N
C o n ten t C o n te n t
99.0 per cent to 100.5 per cent of A IK C S O ^ 12H 20 . 95.0 p er cent to 101.0 per cent.
CHARACTERS CHARACTERS
A p p e a ra n c e A p p e a ra n c e
Granular powder or colourless, transparent, crystalline W hite or slightly yellow, crystalline powder or colourless
masses. crystals, deliquescent
S olubility S o lu b ility
Freely soluble in water, very soluble in boiling water, soluble Very soluble in water, freely soluble in ethanol (96 per cent),
in glycerol, practically insoluble in ethanol (96 per cent). soluble in glycerol.
ID E N T IF IC A T IO N ID E N T IF IC A T IO N
A. Solution S (see Tests) gives the reactions o f sulfates A. Dilute 0.1 m L of solution S2 (see Tests) to 2 m L with
(2.3.1). water R. T he solution gives reaction (a) o f chlorides (2.3.1).
B. Solution S gives the reaction of aluminium (2.3.1). B. Dilute 0.3 m L o f solution S2 to 2 m L with water R.
C. Shake 10 m L o f solution S with 0.5 g o f sodium hydrogen T he solution gives the reaction o f aluminium (2.3.1).
carbonate R and filter. T he filtrate gives reaction (a) o f TESTS
potassium (2.3.1). S o lu tio n S I
TESTS Dissolve 10.0 g in distilled water R and dilute to 100 m L w ith
S o lu tio n S the same solvent.
Dissolve 2.5 g in water R and dilute to 50 m L with the same S o lu tio n S 2
solvent Dilute 50 m L o f solution SI to 100 m L with water R.
A p p e a ra n c e o f so lu tio n A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and colourless (2.2.2, Method IT). Solution S2 is clear (2.2.1) and no t more intensely coloured
p H (2.2.3) than reference solution B 7 (2.2.2, Method II).
3.0 to 3.5. S u lfates (2.4.13)
Dissolve 1.0 g in carbon dioxide-free water R and dilute to Maximum 100 ppm , determined on solution S I.
10 m L with die same solvent. Ir o n (2.4.9)
A m m o n iu m (2.4.1) Maximum 10 ppm , determined on solution S i.
Maximum 0.2 per cent. A lkali a n d alk alin e -e a rth m e ta ls
T o 1 m l, o f solution S add 4 m L of water R. Dilute 0.5 m L Maximum 0.5 per c e n t
o f this solution to 14 m L with water R. T o 20 m L o f solution S2 add 100 m L o f water R and heat to
Iron ( 2. 4. 9) boiling. T o the hot solution add 0.2 m l. of methyl red
Maximum 100 ppm. solution R. Add dilute ammonia R1 until the colour of the
Dilute 2 m L o f solution S to 10 m L with water R. U se in this indicator changes to yellow and dilute to 150 m L with
test 0.3 m L o f thiogfycoUic add R. water R. H eat to boiling and filter. Evaporate 75 m L of the
filtrate to dryness on a water-bath and ignite to constant
H eavy m e ta ls (2.4.8)
mass. T h e residue weighs a maximum of 2.5 mg.
M aximum 20 ppm.
H eav y m e ta ls (2.4.8)
12 m L o f solution S complies with test A. Prepare the
Maximum 20 ppm.
reference solution using lead standard solution (1 ppm Pb) R.
12 m L of solution SI complies w ith test A. Prepare the
ASSAY reference solution using lead standard solution (2 ppm Pb) R.
Dissolve 0.900 g in 20 m L of water R and carry out the
W a te r (2.5.12)
complexometric titration o f aluminium (2.5.11).
42.0 p e r cent to 48.0 per cent, determined on 50.0 mg.
1 m L of 0.1 M sodium edetate is equivalent to 47.44 mg
ofA IK (S 04 ) 2, 12H 20 . A SSAY
Dissolve 0.500 g in 25.0 m L of water R. Carry out the
__________________________________________________________ PhEur
complexometric titration o f aluminium (2.5.11). Titrate with
0.1 M zinc sulfate until the colour o f the indicator changes
from greyish-green to pink. Carry out a blank titration.
1 m L of 0.1 M sodium edetate is equivalent to 24.14 mg
o f A1C13,6H 20 .
1-116 Aluminium Glycinate 2016
T o 10 m L o f solution S add about 0.5 m L o f dilute Place 5 g in a test-tube immersed in ice-water, add 0.4 m L
hydrochloric add R and about 0.5 m L of thioacetamide of a 100 g/L solution of potassium chloride R, 0.1 mT. of
reagent R. N o precipitate is formed. A dd dropwise 5 m L of diphenylandne solution R and, dropwise with sh a k in g , 5 m L of
dilute sodium hydroxide solution R. Allow to stand for 1 h. sulfuric acid R. Transfer the tube to a water-bath at 50 °C.
A gelatinous white precipitate is formed which dissolves upon After 15 min, any blue colour in the solution is no t more
addition o f 5 mT. of dilute sodium hydroxide solution R. intense than that in a standard prepared a t the same time
Gradually add 5 m L of ammonium chloride solution R and and in the same manner using 5 m L of nitrate standard
allow to stand for 30 min. T he gelatinous white precipitate is solution (100 ppm NO?) R.
re-formed. S u lfates (2.4.13)
TESTS M aximum 0.5 p er cent.
S o lu tio n S Dilute 2 m L o f solution S to 20 m L with water R.
A dd 1 g to 4 m L of hydrochloric acid R. H eat at 60 °C for A m m o n iu m (2. 4.1, Method B)
1 h, cool, dilute to 50 m L with distilled water R and filter if M aximum 50 ppm , determined on 1.0 g.
necessary.
Prepare the standard using 0.5 m L of ammonium standard
p H (2.2.5) solution (100 ppm NH 4) R.
5.5 to 8.5.
A rse n ic (2.4.2, Method A)
A d so rp tio n p o w e r M aximum 1 ppm , determined on 1 g.
Dilute the substance to be examined with distilled water R to
Ir o n (2.4.9)
obtain an aluminium concentration o f 5 mg/mL. Prepare
M aximum 15 ppm , determined on 0.67 g.
bovine albumin R solutions with the following concentrations
o f bovine albumin: 0.5 mg/mL, 1 mg/mL, 2 mg/mL, H eav y m e ta ls (2.4.8)
3 mg/mL, 5 m g/m L and 10 mg/mL. I f necessary, adjust the M aximum 20 ppm .
gel and the bovine albumin R solutions to p H 6.0 with dilute Dissolve 2.0 g in 10 m L o f dilute nitric acid R and dilute to
hydrochloric acid R or dilute sodium hydroxide solution R. 20 m L with water R. T h e solution complies with test A.
F or adsorption, mix 1 part o f the diluted gel with 4 parts o f Prepare the reference solution using lead standard solution
each of the solutions o f bovine albumin R and allow to stand (2 ppm Pb) R.
at room tem perature for 1 h. During this time shake the B a c te ria l en d o to x in s (2.6.14)
mixture vigorously at least 5 times. Centrifuge or filter Less than 5 IU of endotoxin per m illigram of alu m in iu m , if
through a non-protein-retaining filter. Immediately determine intended for use in the manufacture of an adsorbed product
the protein content (2.5.33, Method 2) o f either the w ithout a further appropriate procedure for the removal of
supernatant or the filtrate. bacterial endotoxins.
It complies with the test if no bovine alb u m in is detectable in A SSAY
the supernatant or filtrate of the 2 mg/mL bovine albumin R Dissolve 2.50 g in 10 m L o f hydrochloric acid R, heating for
solution (maximum level of adsorption) and in the 30 min at 100 °C on a water-bath. Cool and dilute to 20 m L
supernatant or filtrate o f bovine albumin R solutions o f lower with water R. T o 10 m L o f the solution, add concentrated
concentrations. Solutions containing 3 mg/mL, 5 m g/m L and ammonia R until a precipitate is obtained. Add the smallest
10 mg/mL bovine albumin R may show bovine albumin in the quantity o f hydrochloric acid R needed to dissolve the.
supernatant or filtrate, proportional to the am ount o f bovine precipitate and dilute to 20 m L with water R. Carry out the
albumin in the solutions. complexometric titration o f alu m in ium (2.5.11). Carry out a
S e d im e n ta tio n blank titration.
If necessary, adjust the substance to be examined to p H 6.0
STORA GE
using dilute hydrochloric add R or dilute sodium hydroxide
At a temperature not exceeding 30 °C. D o not allow to
solution R. D ilute with distilled water R to obtain an
freeze. If the substance is sterile, store in a sterile, airtight,
aluminium concentration of approximately 5 mg/mL. I f the
tam per-proof container.
aluminium content of the substance to be e x am in e d is lower
than 5 mg/m L, adjust to p H 6.0 and dilute with a 9 g/L LABELLING
solution o f sodium chloride R to obtain an aluminium T h e label states the declared content o f alu m in iu m .
concentration o f about 1 mg/mL. After shaking for at least -------------------:____________________________________________ PhEur
30 s, place 25 m L of the preparation in a 25 m L graduated
cylinder and allow to stand for 24 h.
It complies with the test if the volume o f the clear
supernatant is less than 5 m L for die gel with an aluminium
content o f about 5 mg/mL.
Dried Aluminium Hydroxide *****
*★ ★*
It complies with the test if the volume o f the clear (Hydrated Aluminium Oxide, *
supernatant is less than 20 m L for the gel with an aluminium Ph Eur monograph 0311)
content o f about 1 mg/mL.
A ctio n a n d u se
C h lo rid e s (2.4.4)
Antacid.
Maximum 0.33 per cent.
P re p a ra tio n s
Dissolve 0.5 g in 10 m L of dilute nitric add R and dilute to
Aluminium Hydroxide Oral Suspension
500 m L w ith water R.
Chewable Aluminium Hydroxide Tablets
N itra te s
Chewable Compound M agnesium Trisilicate Tablets
M axim um 100 ppm.
Co-magaldrox Oral Suspension
Co-magaldrox Tablets
1-118 Aluminium Magnesium Silicate 2016
PhEur.
1.0 g complies with limit test A.
H e a v y m e ta ls (2.4.8)
D E F IN IT IO N
M axim um 20 ppm.
C o n te n t
94.0 per cent to 102.0 per cent of A1P0 4 (Mt 122.0) (ignited Dissolve 1.0 g in dilute hydrochloric add R and dilute to
substance). 20 m L with the same ad d . 12 m L o f the solution complies
with test A. Prepare the reference solution using lead standard
CHARACTERS solution (1 ppm Pb) R.
A p p e a ra n c e
L o ss o n ig n itio n
W hite or alm ost white powder.
10.0 p er cent to 20.0 per cent, determ ined on 1.000 g at
S o lu b ility 800 ± 50 °C.
Very slightly soluble in water, practically insoluble in ethanol
N e u tra lisin g c a p a c ity
(96 per cent). It dissolves in dilute solutions o f mineral ad d s
A dd 0.50 g to 30 m L of 0.1 M hydrochloric add previously
and alkali hydroxides.
heated to 37 °C and maintain at this tem perature for 15 min
ID E N T IF IC A T IO N while stirring. T h e p H (2.2.3) o f the mixture after 15 m in at
A. Solution S (see Tests) gives reaction (b) of phosphates 37 °C is 2.0 to 2.5.
(2.3.1). A SSA Y
B. Solution S gives the reaction o f aluminium (2.3.1). Dissolve 0.400 g in 10 m L o f dilute hydrochloric add R and
TESTS dilute to 100.0 m L with water R. T o 10.0 m L o f the
S o lu tio n S solution, add 10.0 m L o f 0.1 M sodium edetate and 30 m L of
Dissolve 2.00 g in dilute hydrochloric acid R and dilute to a mixture o f equal volumes o f ammonium acetate solution R
100 m L with the same ad d . and dilute acetic add R Boil for 3 min, then cool. A dd 25 m L
o f ethanol (96 per cent) R and 1 m L o f a freshly prepared
A p p e a ra n c e o f so lu tio n
0.25 g/L solution o f dithizone R in alcohol R. T itrate the
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
excess of sodium edetate with 0.1 M zinc sulfate until the
p H (2.2.3) colour changes to pink.
5.5 to 7.2 1 m L o f 0.1 M sodium edetate is equivalent to 12.20 m g o f
Shake 4.0 g with carbon dioxide-free water R and dilute to AIPO 4.
100 m L with the same solvent.
STORA GE
C h lo rid e s (2.4.4)
In an airtight container.
M axim um 1.3 per c e n t
PhEur
Dissolve 50.0 m g in 10 m L o f dilute nitric add R and dilute
to 200 m L with water R.
S o lu b le p h o sp h a te s
M axim um 1.0 per cent, calculated as PO 43-. ,* * * ★
★
Test solution Stir 5.0 g with 150 m L o f water R for 2 h. Filter Aluminium Phosphate Gel ★ ★
and wash the filter with 50 m L o f water R. Com bine the (Ph Eur monograph 2166) * * * * *
filtrate and the washings and dilute to 250.0 m L with
water R D ilute 10.0 m L o f this solution to 100.0 m L with A c tio n a n d u se
water R. A ntad d ; vacdne adjuvant
Reference solution (a) Dissolve 2.86 g o f potassium dihydrogen
PhEur-----------------------------------------------------------------------------------------
phosphate R in water R and dilute to 100 m L with th e same
solvent D E F IN IT IO N
Reference solution (b) Dilute 1 m L o f reference solution (a) to H ydrated A1P0 4 in g d form.
5 m L with water R. C o n te n t
Reference solution (c) Dilute 3 m L o f reference solution (a) to 19.0 per cent to 21.0 per cent o f AIPO 4.
5 m L with water R.
CHARACTERS
T reat each solution as follows. T o 5.0 m L add 4 m L o f dilute A p p e a ra n c e
sulfuric add R, 1 m L of ammonium mdybdate solution R, 5 m L Gel.
o f water R and 2 m L o f a solution containing 0.10 g of
S o lu b ility
4-methylaminophenol sulfate R, 0.5 g o f anhydrous sodium
Practically insoluble in water, in ethanol (96 per cent) and in
sulfite R and 20.0 g o f sodium metabisulfite R in 100 m L of
methylene chloride. It dissolves in dilute solutions o f mineral
water R Shake and allow to stand for 15 min. Dilute to
ad d s.
25.0 m L with water R and allow to stand for a further
2016 Aluminium Powder 1-121
add R. D ilute 10.0 m L of the solution to 25.0 m L with 1.0 m L of lanthanum chloride solution R and dilute to
water R. 50.0 m L with water R.
Lead Reference solutions Into 5 separate 50 m L volumetric flasks,
M axim um 5 ppm . introduce respectively 1.0 mL, 2.5 m l , 5.0 ml-, 7.5 m L and
10.0 m L o f aluminium standard solution (100 ppm Al) R, add
Atomic absorption spectrometry (2.2.23, Method I).
1 m L of lanthanum chloride solution R and 10 m L o f the blank
Test solution T ransfer 5.0 g to a 250 m L beaker containing solution, and dilute to 50.0 m L with water R.
50 m L o f dilute hydrochloric add R Mix, cover with a watch
glass an d boil for 15 min. Allow to cool to room Source Aluminium hollow-cathode lamp.
temperature. Centrifuge, and decant the supernatant through Wavelength 309.3 nm .
a rapid-flow filter paper into a 250 m L beaker. T o the Atomisation device Acetylene-nitrous oxide flame.
insoluble m atter add 25 m L of hot water R. Stir vigorously, Sodium
centrifuge, and decant the supernatant through the same Atomic emission spectrometry (2.2.22, Method i).
filter into the beaker. Repeat the extraction with 2 additional
Test solution T o 2.0 m L o f solution A, prepared in the assay
quantities, each o f 25 mL, o f hot water R, decanting each
o f aluminium, add 1 m L of a 12.5 g/L solution o f caesium
supernatant through the filter into the beaker. W ash the filter
chloride R and dilute to 20.0 m L with water R.
with 25 m L o f h o t water R, collecting die filtrate in the
beaker. C oncentrate the combined filtrates by gently boiling Reference solutions Into 5 separate 200 m L volumetric flasks,
to about 15 mT. Add about 0.05 m L o f heavy metal-free nitric each containing 10 m L o f a 12.5 g/L solution of caesium
add R, heat to boiling and allow to cool to room chloride R, introduce respectively 1.0 m l, 2.0 m l , 4.0 m l,
temperature. Filter the concentrated extracts through a rapid- 6.0 m L and 10.0 m L o f sodium standard solution
flow filter paper into a 25 m L volumetric flask. Transfer the (200 ppm Na) R and dilute to 200.0 m L with water R.
rem aining contents o f the beaker through the filter paper and Wavelettgth 589.0 nm .
into die volum etric flask with water R and dilute to 25.0 m L __________________________________________________________ PhEur
with the sam e solvent.
Reference solutions Into 4 separate 100 m L volumetric flasks,
introduce respectively 3.0 mL, 5.0 mL, 10.0 m L and
15.0 m L o f lead standard solution (10 ppm Pb) R, add
0.20 mT. o f heavy metal-free nitric add R and dilute to Aluminium Stearate *****
100.0 m L w ith water R.
(Ph Eur monograph 1663) *
Source L ead hollow-cathode lamp.
PhEur__________________________________________________________
Wavelength 217.0 nm.
D E F IN IT IO N
Atomisation device Air-acetylene flame.
Aluminium salts o f a mixture of solid organic a d d s consisting
L oss o n d ry in g (2.2.32) mainly o f variable proportions o f aluminium stearate and
M axim um 8.0 per cent, determ ined on 1.000 g by drying in aluminium palmitate. T h e organic adds are obtained from
an oven at 105 °C for 4 h. sources o f vegetable o r animal origin.
L oss o n ig n itio n C o n te n t
5.0 per cent to 11.0 per cent (dried substance), determ ined — aluminium (Al; A t 26.98): 3.0 per cent to 9.0 per cent
on 1.000 g by ignition in a platinum crucible to constant (dried substance);
mass a t 1000 ± 25 °C. — stearic add in the fatty add fraction: m inim um
M ic ro b ia l c o n ta m in a tio n 40.0 per cent;
T A M C : acceptance criterion 103 CFU /g (2.6.12). — sum of stearic acid and palmitic add in the fatty add fraction:
T Y M C : acceptance criterion 102 C FU /g (2.6.12). m inim um 90.0 per cent.
D. 1 m L o f solution S gives the reaction of aluminium Reference solution Prepare a solution containing
(2.3.1). T he addition of 0.5 m L o f dilute hydrochloric add R 0.00165 ng/mL o f cadmium nitrate tetrahydrate R in the blank
described in the general m ethod is omitted. solution (equivalent to 0.006 pg/m L o f Cd).
TESTS D ilute 1.0 m L o f the test solution to 10.0 m L with the blank
S o lu tio n S solution. Prepare mixtures of this solution, the reference
T o 5.0 g add 50 m L o f peroxide-free ether R, 20 m L o f dilute solution and the blank solution in the following proportions:
nitric add R and 20 m L of distilled water R and heat gently (1.0:0:1.0 V/V/V), (1.0:0.25:0.75 V/V/V),
under a reflux condenser until dissolution is complete. Allow (1.0:0.5:0.5 V/V/V), (1.0:0.75:0.25 V/V/V). T o each mixture
to cool. In a separating funnel, separate the aqueous layer add 50 )iL o f the modifier solution and mix. These solutions
and shake the ether layer with 2 quantities, each o f 4 mT, o f contain respectively 0 jxg, 0.0015 jig, 0.0030 ng and
distilled water R. Com bine the aqueous layers, wash with 0.0045 ng o f cadmium per millilitre from the reference
15 m L of peroxide-free ether R and dilute to 50.0 m L with solution. K eep the remaining test solution for use in the test
distilled water R (solution S). Evaporate the ether layer to for lead and nickel.
dryness and dry the residue at 100-105 °C. Keep the residue Source Cadm ium hollow-cathode lamp.
for identification tests A and B. Wavelength 228.8 nm .
A cid ity o r alk a lin ity Atomisation device Furnace.
T o 1.0 g add 20 m L o f carbon dioxide-free water R and boil Platform Pyrolytically coated with integrated tube.
for 1 min with continuous sh ak ing. Cool and filter.
Operating conditions U se the tem perature programme
T o 10 m L o f the filtrate add 0.05 m L of bromothymol blue
recom m ended for cadmium by the GFAA manufacturer.
solution R4. N o t m ore than 0.05 m L o f 0.1 M hydrochloric A n example o f tem perature parameters for GFAA analysis of
add or 0.1 M sodium hydroxide is required to change the
cadm ium is shown below.
colour of the indicator.
C h lo rid es (2.4.4)
Stage Final temperature Ramp time Hold time
M aximum 0.1 per cent.
CC) (s) (s)
Dilute 0.5 m L o f solution S to 15 m L with water R. Drying 110 10 20
S u lfates (2.4.13)
Ashing 600 10 30
M aximum 0.5 per cent.
Atomisation 1800 0 5
Dilute 0.3 m L o f solution S to 15 m L with distilled water R.
C a d m iu m
M axim um 3 ppm . L ead
Atomic absorption spectrometry (2.2.23, Method II). M axim um 10 ppm .
For the preparation of all aqueous solutions and for the rinsing of Atomic absorption spectrometry (2.2.23, MethodII).
glassware before use, employ water that has been passed through a For the preparation of aU aqueous solutions and for the rinsing of
strong-acid, strong-base, mixed-bed ion-exchange resin before use. glassware before use, employ water that has been passed through a
Select all reagents to have as low a content of cadmium, lead and strong-acid, strong-base, mixed-bed ion-exchange resin before use.
nickel as practicable and store all reagent solutions in containers of Select all reagents to have as low a content of cadmium, lead and
borosUicate glass. Clean glassware before use by soaking in warm nickel as practicable and store aU reagent solutions in containers of
8 M nitric acid for 30 min and by rinsing with deionised water. borosUicate glass. Clean glassware before use by soaking in warm
Blank solution D ilute 25 m L o f cadmium- and lead-free nitric 8 M nitric add for 30 min and by rinsing with deionised water.
acid R to 100.0 m L w ith water R. Blank sdution U se the solution described in the test for
Modifier solution Dissolve 20 g o f ammonium dihydrogen cadmium.
phosphate R and 1 g o f magnesium nitrate R in water R and Modifier solution U se the solution described in the test for
dilute to 100 m L with the same solvent. Alternatively, use an cadmium.
appropriate matrix modifier as recommended by the graphite Test solution U se the solution described in the test for
furnace atom ic absorption (GFAA) spectrometer cadmium.
manufacturer.
Reference sdution Prepare a solution of 0.100 |ig/mL of Pb by
Test solution Place 0.100 g of the substance to be examined in suitable dilutions o f lead standard solution (100 ppm Pb) R
a polytetrafluoroethylene digestion bom b and add 2.5 m L o f with the blank solution.
cadmium- and lead-free nitric acid R. Close and seal th e bom b
Prepare mixtures o f the test solution, the reference solution
according to the m anufacturer's operating instructions. When
and the blank solution in the following proportions:
using a digestion bomb, be thoroughly familiar with the safety and
(1.0:0:1.0 V/V/V), (1.0:0.5:0.5 V/V/V), (1.0:1.0:0 V/V/V).
operating instructions. Carefully follow the bomb manufacturer's
T o each mixture add 50 pL o f the modifier solution and mix.
instructions regarding care and maintenance of these digestion
These solutions contain respectively 0 |ig5 0.025 |ig and
bombs. Do not use metal-jacketed bombs or liners that have been
0.05 jig o f lead p er millilitre from the reference solution.
used with hydrochloric acid due to contamination from corrosion of
the metal jacket by hydrochloric add. H eat the bom b in an oven Source L ead hollow-cathode lamp.
at 170 °C for 3 h. Cool the bom b slowly in air to room Wavelength 283.3 nm .
tem perature accord in g to the bom b m anufacturer's Atomisation device Furnace.
instructions. Place the bom b in a fume cupboard and open Platform Pyrolytically coated with integrated tube.
carefully as corrosive gases may be expelled. Dissolve die
Operating conditions Use the tem perature programme
residue in water R and dilute to 10.0 m L with the same
recom m ended for lead by the GFAA manufacturer.
solvent.
A n example of tem perature parameters for GFAA analysis of
lead is shown below.
2016 Aluminium Stearate 1-125
Stage Final temperature Ramp time Hold time solution for 30 m in under reflux on a w ater-bath, swirling
_________________CQ___________ &__________ isi__ frequently. Allow to cool. Add 100 m L o f water R and adjust
Drying 110 10 20 to about p H 1 by adding approximately 12 m L of dilute
Ashing 450 10 30 sodium hydroxide solution R. Add 20.0 m L o f 0.1 M sodium
edetate and adjust to between pH 5 and p H 6 by the addition
Atomisation 2000 0 5
of sodium acetate R. Add 70 mg of xylenol orange triturate R
and titrate immediately and quickly with 0.1 M zinc sulfate
until the colour changes from yellow to pinkish-violet.
Nickel
M axim um 5 ppm . 1 m L of 0.1 M sodium edetate is equivalent to 2.698 mg of
Atomic absorption spectrometry (2.2.23, Method II). Al.
For the preparation of all aqueous solutions and for the rinsing of S te a ric a c id a n d p a lm itic acid
glassware before me, employ water that has been passed through a Gas chromatography (2.2.28): use the normalisation
strong-acid, strong-base, mixed-bed ion-exchange resin before use. procedure.
Select al1 reagents to have as low a content of cadmium, lead and Test solution In a conical flask fitted with a reflux condenser,
nickel as practicable and store al1 reagent solutions in containers of dissolve 0.100 g o f the substance to be examined in 5 m L o f
borosUicate glass. Clean glassware before use by soaking in warm boron trifliioride-methanol solution R. Boil under a reflux
8 M nitric acid for 30 mm and by rinsing with deionised water. condenser for 10 min. Add 4 m L of heptane R through the
Blank solution U se the solution described in the test for condenser and boil again under a reflux condenser for
cadmium. 10 min. Allow to cool. A dd 20 m L of saturated sodium
chloride solution R. Shake and allow the layers to separate.
Modifier solution Dissolve 20 g of ammonium dihydrogen
D ry the organic layer over 0.1 g of anhydrous sodium sulfate R
phosphate R in water R and dilute to 100 m L with the same
previously washed with heptane R. D ilute 1.0 m L of the
solvent. Alternatively, use an appropriate matrix modifier as
solution to 10.0 m L with heptane R.
recom m ended by the GFAA spectrometer manufacturer.
Reference solution Prepare the reference solution in the same
Test solution U se the solution described in the test for
m anner as the test solution using 50.0 m g of palmitic
cadmium.
acid CRS and 50.0 mg of stearic acid CRS instead o f the
Reference solution Prepare a solution of 0.050 ng/mL of N i by substance to be examined.
suitable dilutions o f a 0.2477 |ig/mL solution of nickel nitrate
Column:
hexahydrate R with the blank solution.
— material: fused silica;
Prepare mixtures o f the test solution, the reference solution — size. I = 30 m , 0 = 0.32 mm;
and the blank solution in the following proportions: — stationary phase: macrogol 20 000 R (film thickness
(1.0:0:1.0 V/V/V), (1.0:0.5:0.5 VIVIV), (1.0:1.0:0 VIVIV). 0.5 tun).
T o each mixture add 50 (iL of the modifier solution and mix.
Carrier gas helium for chromatography R.
T hese solutions contain respectively 0 ng, 0.0125 fog and
0.025 ng o f nickel per millilitre from the reference solution. Flow rate 2.4 ml 7min.
Source Nickel hollow-cathode lamp. Temperature:
Wavelength 232.0 nm.
Time Temperature.
Atomisation device Furnace.
(min) rc)
Platform Pyrolytically coated with integrated tube. Column 0 -2 70
Operating conditions Use the temperature programme
2 -3 6 70 240
recom m ended for nickel by the GFAA manufacturer.
A n example o f tem perature parameters for GFAA analysis of 36-41 240
nickel is shown below. Injection port 220
Detector 260
Stage Final temperature Ramp time Hold time
CC) (») (s)
Drying 110 10 20 Detection Flam e ionisation.
Ashing 1000 20 30 Injection 1 (iL.
Atomisation 2300 0 5 Relative retention W ith reference to methyl stearate: methyl
palmitate = about 0.9.
System suitability: reference solution:
L oss o n d ry in g ( 2.2.32) — resolution: minimum 5.0 between the peaks due to methyl
M axim um 6.0 per cent, determined on 1.000 g by drying in palmitate and methyl stearate;
an oven at 105 °C. — repeatability: maximum relative standard deviation of
M ic ro b ia l c o n ta m in a tio n 3.0 per cent for the areas of the peaks due to methyl
T A M C : acceptance criterion 103 C FU/g (2.6.12). palmitate and methyl stearate after 6 injections; maximum
relative standard deviation of 1.0 per cent for the ratio of
T Y M C : acceptance criterion 102 CFU/g (2.6.12).
the areas of the peaks due to methyl palm itate to the
Absence of Escherichia colt (2.6.13). areas of the peaks due to methyl stearate after
Absence of Salmonella (2.6.13). 6 injections.
A SSA Y __________________________________________________________ PhEur
A lu m in iu m
T o 0.250 g in a 250 m L conical flask add 20 m L of
methanol R and, slowly, 2 m L o f sulfuric acid R. H eat the
1-126 Aluminium Sulfate 2016
★ ★ STO RA G E
Aluminium Sulfate ★ ★ In an airtight container.
Aluminium Sulphate *****
PhEur
(Ph Eur monograph 0165)
A12(S04)3îxH20 342.1
(anhydrous substance)
★ ★
Alverine Citrate ★ ★
P re p a r a tio n *****
Aluminium Acetate E ar D rops
(Ph Eur monograph 2156)
Ph E n _______________________________________ CO j H
ch3
D E F IN IT IO N CO2H
C o n te n t OH
51.0 per cent to 59.0 per cent of A12(S 04 )3. COjH
It contains a variable quantity of water of crystallisation.
CHARACTERS C 26H 35NO 7 473.6 5560-59-8
A p p e a ra n c e
A c tio n a n d u se
Colourless, lustrous crystals or crystalline masses.
Sm ooth muscle relaxant; antispasmodic.
S o lu b ility
P re p a r a tio n
Soluble in cold water, freely soluble in hot water, practically
Alverine Capsules
insoluble in ethanol (96 per cent).
ID E N T IF IC A T IO N PhEur_____________________________________
Time Temperature
(min) CC)
Column 0 -7 120
24-39 290
Detector 290
E. 3-phenyl-AT^ST-bis(3-phenylpropyl)propan-l-amine.
Detection Flame ionisation.
PtiEur
Irtjection 1 pL.
Identification of impurities Use the chromatogram supplied
with alverine for peak identification CRS and the
chromatogram obtained with reference solution (c) to identify
the peaks due to impurities C and E. Amantadine Hydrochloride ***\
Relative retention W ith reference to alverine (retention
time = about 16 m in): impurity A = about 0.28; (Ph Em monograph 0463) *
impurity B = about 0.29; impurity C = about 0.46;
impurity D = about 0.97; impurity E = about 1.7.
System suitability: reference solution (a):
— resolution: m inim um 3.0 between the peaks due to
impurity D and alverine.
C 10H 18CIN 187.7 665-66-7
Limits:
— impurities A , B: for each impurity, maximum 0.1 per cent; Action and use
— impurity C: maxim um 0.2 per cent; Viral replication inhibitor (influenza A); dopamine receptor
— impurities D, E: for each impurity, maximum 0.3 per cent; agonist; treatm ent of influenza and Parkinson’s disease.
— unspecified impurities: for each impurity, maximum
0.10 per cent; Preparations
Amantadine Capsules
— total: maximum 1.0 per cent;
— disregard limit, the area o f the principal peak in the A m antadine Oral Solution
chrom atogram obtained with reference solution (b)
PhEir _______________________ :___________________________________
(0.05 per cent).
H eav y m e ta ls (2.4.8)
DEFINITION
Maximum 20 ppm . Tricyclo[3.3.1. l 3,7]decan-l-am ine hydrochloride.
.OH
Identification of impurities U se the chrom atogram obtained
with reference solution (b) to identify the peak due to
impurity B.
»-.X )
Relative retention W ith reference to ambroxol (retention C. trans-4- [[(£)-2-am ino-3,5-dibromobenzyliden] amino]
time = about 9 min): impurity B = about 0.6.
cyclohexanol.
System suitability: reference solution (b):
— resolution: minimum 4.0 between the peaks due to .OH
impurity B and ambroxol.
Limits: . - „ O
H
— unspecified impurities: for each impurity, not more than the
area of the principal peak in the chromatogram obtained
D . os-4- [(2-amino-3,5-dibromobenzyl)amino] cyclohexanol,
with reference solution (a) (0.10 per cent),
— total: not m ore than 3 times the area of the principal peak E. A r-C H = 0 : 2-amino-3,5-dibromobenzaldehyde.
in the chromatogram obtained with reference solution (a) PhEtr
(0.3 per cent),
— disregard limit. 0.5 times the area o f the principal peak in
the chromatogram obtained with reference solution (a)
(0.05 per cent). ★*★
★ ★
H eavy m e ta ls (2.4.8) Amfetamine Sulfate ★ ★
M aximum 20 ppm. Amfetamine Sulphate * * * * *
1.0 g complies with test C. Prepare the reference solution (Ph Eur monograph 0368)
using 2 m L o f lead standard solution (10 ppm Pb) R.
L oss o n d ry in g (2.2.32)
M aximum 0.5 per cent, determined on 1.000 g by drying in H2SO4 and enantiomer
an oven at 105 °C.
S u lfa te d a s h (2.4.14)
M aximum 0.1 per cent, determined on 1.0 g. C 18H 28N 2O 4S 368.5 60-13-9
ASSAY
A c tio n a n d u se
Dissolve 0.300 g in 70 m L of ethanol (96 per cent) R and add
Releases dopamine; central nervous system stimulant.
5 m L of 0.01 M hydrochloric acid. C a n y out a potentiometric
titration (2.2.20), using 0.1 M sodium hydroxide. Read PhEur__________________________________________________________
the volume added between the 2 points o f inflexion.
D E F IN IT IO N
1 m L of 0.1 M sodium hydroxide is equivalent to 41.46 mg of
Bis [(2.&S)-1-phenylpropan-2-amine] sulfate.
C 13Hj9Br2C lN 20 .
C o n te n t
STORA GE
99.0 per cent to 100.5 per cent (dried substance).
Protected from light.
CHARACTERS
IM P U R IT IE S
A p p e a ra n c e
Other detectable impurities (the following substances would, if W hite or alm ost white powder.
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general S o lu b ility
acceptance criterion for other/unspecified impurities and/or Freely soluble in water, slightly soluble in ethanol
by the general monograph Substances for pharmaceutical use (96 per cent).
(2034). It is therefore no t necessary to identify these ID E N T IF IC A T IO N
impurities for demonstration o f compliance. See also 5.10. First identification A , B, E.
Control of impurities in substances for pharmaceutical use): A, B, Second identification A, C, D, E.
C, D, E.
A. Optical rotation (2.2.7): —0.04° to + 0.04° (measured in a
2 dm tube), determined on solution S (see Tests).
B. Infrared absorption spectrophotometry (2.2.24).
Ar- =
Preparation Mulls in liquid paraffin R.
Br Comparison Ph. Eur. reference spectrum of amfetamine sulfate.
C. T o 50 m L of solution S add 5 m L o f strong sodium
A. A r-C H 2O H : (2-amino-3,5-dibromophenyI)methanol, hydroxide solution R and 0.5 m l. o f benzoyl chloride R and
shake. Continue to add benzoyl chloride R in portions of
0.5 m L until no further precipitate is form ed Filter, wash
the precipitate with water R, reciystallise twice from a
mixture o f equal volumes o f ethanol (96 per cent) R and
w
water R, then dry at 100-105 °C. T h e crystals m elt (2.2.14)
at 131 °C to 135 °C.
Br D . T o about 2 m g add 1 m L o f sulfuric acid-formaldehyde
reagent R. A n orange colour develops and quickly becomes
B. trans-4-(6,8-dibrom o-1,4-dihydroquinazolin-3 (2H)-yl) dark-brown.
cyclohexanol, E. Solution S gives reaction (a) o f sulfates (2.3.1).
2016 Amidotrizoic Acid 1-131
System suitability. through a sintered-glass filter (2.1.2) and wash the filter with
— resolution: m inim um 1.5 between the peaks due to several quantities o f water R. Collect the filtrate and
impurities B and C in the chromatogram obtained with washings. Add 40 m L o f dilute sulfuric acid R and titrate
reference solution (c); immediately with 0.1 M silver nitrate. D eterm ine the
— signal-to-noise ratio: minimum 25 for the principal peak in end-point potentiometrically (2.2.20), using a suitable
the chromatogram obtained with reference solution (b). electrode system such as silver/mercurous sulfate.
Limits: 1 m L of 0.1 M silver nitrate is equivalent to 20.47 m g of
— impurity B: not m ore than the area of the principal peak C 11H 9I 3N 2O 4.
in the chromatogram obtained with reference solution (a)
STO RA G E
(0.1 per cent);
Protected from light.
— impurities A , D: for each impurity, not more than the area
of the principal peak in the chromatogram obtained with IM P U R IT IE S
reference solution (b) (0.01 per cent); Specified impurities A, B, D
— unspecified impurities: for each impurity, not more than Other detectable impurities (the following substances would, if
0.5 times the area o f the principal peak in the present at a sufficient level, be detected by one or other o f
chrom atogram obtained with reference solution (a) the tests in the monograph. T hey are limited by the general
(0.05 per cent); acceptance criterion for other/unspecified impurities and/or
— total: not m ore than 1.5 times the area of the principal by the general m onograph Substances for pharmaceutical use
peak in the chromatogram obtained with reference (2034). It is therefore not necessary to identify these
solution (a) (0.15 per cent); impurities for dem onstration of compliance. See also 5.10.
— disregard limit. 0.3 times the area o f the principal peak in Control of impurities in substances for pharmaceutical use): C, E.
the chromatogram obtained with reference solution (a)
(0.03 per cent), except for the peaks due to impurities A CO2H
and D.
H a lid e s ex p ressed a s ch lo rid es (2.4.4)
M aximum 150 ppm . h2n
Dissolve 0.55 g in a mixture o f 4 m L o f dilute sodium
hydroxide solution R and 15 m L of water R. Add 6 m l. of
dilute nitric acid R and filter. A. 3-(acetylamino)-5-amino-2,4,6-triiodobenzoic acid,
F re e a ro m a tic a m in e s
Maintain the solutions and reagents in iced water, protected from c o 2h
Identification of impurities Use the chromatogram supplied Calculate the percentage content o f C 22H 43N 5O 13 taking into
with amikacin for system suitability CRS and the account the assigned content o f armkadn CRS.
chromatogram obtained with reference solution (c) to identify IM P U R IT IE S
the peaks due to impurities A, B, F and H ; use the
Spedfied impurities A, B, F, H , I
chromatogram obtained with reference solution (d) to
identify the peak due to impurity I. Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one o r other of
Relative retention W ith reference to amikacin (retention
the tests in the monograph. T hey are limited by the general
time = about 28 min): impurity I = about 0.13;
acceptance criterion for other/unspecified impurities and/or
impurity F = about 0.92; impurity B = about 0.95;
by the general monograph Substances for pharm aceutical use
impurity A = about 1.62; impurity H = about 1.95.
(2034). It is therefore not necessary to identify these
System suitability: reference solution (c): impurities for demonstration o f compliance. See also 5.10.
— peak-tchvalley ratio: minimum 5, where Hp = height above Control of impurities in substances for pharmaceutical use): C, D,
the baseline of the peak due to im purity B and E, G.
Hv = height above the baseline o f the lowest point o f the
curve separating this peak from the peak due to amikacin;
if necessary, adjust the volume of tetrahydrofuran in the
mobile phase.
Calculation ofpercentage contents:
— for im purity I, use the concentration of impurity I in
reference solution (d);
— for impurities other than I, use the concentration o f
amikacin in reference solution (a).
Limits:
— impurities A , B, F, H} I: for each impurity, maximum
0.5 per cent;
— any other impurity: for each impurity, maximum
0.5 per cent; A. 4-0-(3-amino-3-deoxy-q-D-glucopyranosyl)-6-0-(6-amino-
— total: maximum 1.5 per cent; 6 -deoxy-a-D-glucopyranosyi)-1-N- [(2S)-4-amino-2-
— reporting threshold: 0.1 per cent. hydroxybutanoyl] -2-deoxy-L-streptamine,
W a te r (2.5.12)
M aximum 8.5 per cent, determined on 0.200 g.
S u lfa te d a s h (2.4.14)
Maximum 0.5 per cent, determined on 1.0 g.
A SSAY
Liquid chromatography (2.2.29).
Test solution Dissolve 50.0 m g o f the substance to be
examined in the mobile phase and dilute to 10.0 m L with
the mobile phase.
Reference solution Dissolve 50.0 mg o f amikacin CRS in the
mobile phase and dilute to 10.0 m L with the mobile phase.
Column: B. 4-0-(3-amino-3-deoxy-ce-D-glucopyranosyl)-6-0-(6-amino-
— size. I = 0.25 m , 0 = 4.6 mm; 6-deoxy-a-D-glucopyranosyl)-1,3-N-bis [(2S)-4-amino-2-
— stationary phase, end-capped octadecylsüyl silica gelfor hydroxybutanoyl] -2-deoxy-L-streptamine,
chromatography R (5 nm);
— temperature: 40 °C.
Mobile phase A mixture prepared with carbon dioxide-free
water R, containing 1.8 g/L o f sodium octanesulfonate R,
20 g/L of anhydrous sodium sulfate R1, 5.8 per cent V/V of
acetomtrüe R l, and 5 p er cent V/V o f 0.2 M potassium
dihydrogen phosphate R previously adjusted to p H 3.0 with
dilute phosphoric add R; degas.
Flow rate 1.0 mlVmin.
Detection Spectrophotom eter at 200 nm . *
Irtjection 20 |1L. C . 4-0-(6-amino-6-deoxy-cc-D-glucopyranosyl)-6-0-[3-[[(2S)-
Run time 1.3 times the retention time o f amikacin. 4-amino-2-hydroxybutanoyi] amino] -3-deoxy-a-D-
Retention time Amikacin = about 30 min. glucopyranosyl] -2-deoxy-D-streptamine,
System suitability: reference solution:
— symmetry factor, maximum 1.5 for the peak due to
amikacin; if necessary, adjust the am ount o f acetomtrüe R l
in the mobile phase; peak splitting may be observed w hen
the retention time becomes too short;
— repeatability: maximum relative standard deviation of
1.5 per cent after 6 injections.
2016 Amikacin Sulfate 1-135
HOjC NH,
H OH
I. (25)-4-amino-2-hydroxybutanoic acid.
PhEur
★ ★
E. 4-0-(3-amino-3-deoxy-a-D-glucopyranosyl)-6-0- [6-[[(25)- Amikacin Sulfate ★ ★
4-amino-2-hydroxybutanoyl] amino] -6-deoxy-a-D- *****
Amikacin Sulphate
glucopyranosyl] -2-deoxy-L-streptamine3
(Ph. Eur. monograph 1290)
PhEtr_____________________________
D E F IN IT IO N
6-0-(3-Amino-3-deoxy-a-D-glucopyranosyl)-4-0-(6-amino-6-
deoxy-a-i>glucopyranosyl)-l-N-[(25)-4-amino-2-
hydroxybutanoyl] -2-deoxy-D-strq)tamine sulfate.
Antimicrobial substance obtained from kanamycin A.
Semi-synthetic product derived from a fermentation product.
G . 6 -0 - (3-amino-3-deoxy-a-D-glucopyranosyl)-4-0-
(6-amino-6-deoxy-a-D-glucopyranosyl)-l-N-[(2R)-4-amino-2- C o n te n t
hydroxybutanoyl] -2-deoxy-D-streptamine, 96.5 per cent to 102.0 per cent (dried substance).
CHARACTERS
A p p e a ra n c e
White or almost white powder.
Solubility
Freely soluble in water, practically insoluble in acetone and
in ethanol (96 p er cent).
ID E N T IF IC A T IO N
A. Infrared absorption spectrophotometry (2.2.24).
Comparison amikacin sulfate CRS.
B. Thin-layer chromatography (2.2.27).
1-136 Amikacin Sulfate 2016
Test solution Dissolve 25 mg o f the substance to be examined tetrahydrqfuran R, and 5 per cent V/V o f 0.2 M potassium
in water R and dilute to 10 m L with the same solvent. dihydrogen phosphate R previously adjusted to p H 3.0 with
Reference solution (a) Dissolve 25 m g of amikacin sulfate CRS dilute phosphoric add R; degas;
in water R and dilute to 10 m l. with the same solvent.
Reference solution (b) Dissolve 5 mg o f kanamycin Time Mobile phase A Mobile phase B
monosulfate CRS in 1 m L o f the test solution and dilute to (min) (per cent V/V) (per cent V/V)
10 m L with water R. 0-3 100 0
Amiloride Hydrochloride
(Ph. Eur. monograph 0651) *
O NH
CK .N. JLN J k NH,
H2N
XX H
Y
N NH2
, HCI, 2 H ,0 .
PhEur__________________________________________________________
DEFINITION
3,5-Diarnino-Ar-carbamirnidoyl-6-chloropyrazine-2-
F. 6-0-(3-amino-3-deoxy-a-D-glucopyranosyl)-4-0- [6- [(25)- carboxamide hydrochloride dihydrate.
4-amino-2-hydroxybutanoyl] amino-6-deoxy-a-D-
Content
glucopyranosyQ-l-N- [(25)-4-amino-2-hydroxybutanoyi]-2-
98.0 per cent to 101.0 per cent (anhydrous substance).
deoxy-D -streptam ine}
CHARACTERS
Appearance
Pale yellow or greenish-yellow powder.
Solubility
Slightly soluble in water and in anhydrous ethanol.
IDENTIFICATION
First identification A, D.
Second identification B, C, D.
A. Infrared absorption spectrophotom etry (2.2.24).
Comparison amiloride hydrochloride CRS.
G. 6-0-(3-amino-3-deoxy-a-D-glucopyranosyl)-4-0-
B. Thin-layer chromatography (2.2.27).
(6-araino-6-deoxy-a-D-glucopyranosyl)-l-7V-[(2i?l)-4-ainino-2-
hydroxybutanoyl]-2-deoxy-D-streptamine, Test solution Dissolve 40 m g o f the substance to be examined
in methanol R and dilute to 10 m L w ith the same solvent.
Reference solution Dissolve 40 m g o f amiloride
hydrochloride CRS in methanol R and dilute to 10 m L with the
same solvent.
Plate TLC silica gel plate R.
Mobile phase dilute ammonia R l, water R, dioxan R
(6:6:88 VIV/V); freshly prepared mixture.
Application 5 |iL.
Development Over 2/3 of the plate.
Drying In air.
H . 6-0-(3-amino-3-deoxy-a-D-gJucopyranosyJ)-l-N-[(25)-4- Detection Examine in ultraviolet light at 365 nm.
amino-2-hydroxybutanoyI] -4-0- (2,6-diamino-2, 6-dideoxy-a- Results T h e principal spot in the chrom atogram obtained with
D-gJucopyranosyl)-2-deoxy-D-streptaminej the test solution is similar in position, fluorescence and size
to the principal spot in the chrom atogram obtained w ith the
H° 2 C ^ ^ , N H 2 reference solution.
H OH C. Dissolve about 10 m g in 10 m L o f water R. Add 10 m L
o f a 200 g/L solution of cetrimide R} 0.25 m L of dilute sodium
I. (25)-4-amino-2-hydroxybutanoic add. hydroxide solution R and 1 m L o f bromine water R. A greenish-
yellow colour is produced. Add 2 m L o f dilute hydrochloric
___________________________________________________________ PhEur
acid R. T h e solution becomes deep yellow and shows blue
fluorescence in ultraviolet light at 365 nm.
D. It gives reaction (b) o f chlorides (2.3.1).
2016 Aminobenzoic Acid 1-139
TESTS ASSAY
Free acid Dissolve 0.200 g in a mixture o f 5.0 m L o f 0.01 M
Dissolve 1.0 g in a mixture o f 50 m L o f methanol R and hydrochloric add and 50 m L of ethanol (96 per cent) R. Carry
50 m L o f water R and titrate with 0.1 M sodium hydroxide, out a potentiom etric titration (2.2.20), using 0.1 M sodium
determ ining the end-point potentiometrically (2.2.20). hydroxide. Read the volume added between the 2 points o f
N o t m ore than 0.3 m L o f 0.1 M sodium hydroxide is required inflexion.
to reach the end-point. 1 m L of 0.1 M sodium hydroxide is equivalent to 26.61 m g
Related substances Of C6H 9CI2N 7O.
Liquid chrom atography (2.2.29). STORAGE
Test solution Dissolve 20.0 m g o f the substance to be Protected from light.
examined in a mixture of 1 volume o f acetonitrile R and
IM P U R IT IE S
3 volumes o f water R and dilute to 10.0 m L with the same
mixture o f solvents.
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one o r other o f
Reference solution (a) Dilute 1.0 m L o f the test solution to the tests in the monograph. T hey are limited by the general
100.0 m L with a mixture o f 1 volume o f acetomtrUe R and
acceptance criterion for other/unspecified impurities and/or
3 volumes o f water R. by the general monograph Substances for pharmaceutical use
Reference solution (b) Dilute 1.0 m L o f reference solution (a) (2034). It is therefore not necessary to identify these
to 10.0 m l, with a mixture o f 1 volume o f acetonitrUe R and impurities for demonstration o f compliance. See also 5.10.
3 volumes of water R. Control of impurities in substances for pharmaceutical use): A .
Reference solution (c) Dissolve 5.0 m g o f amHoride
impurity A CRS in a mixture of 1 volume o f acetonitrUe R and
3 volumes of water R and dilute to 5.0 m L with die same ,c h 3
mixture o f solvents. Dilute 1.0 m L o f this solution to
100.0 m L with a mixture of 1 volume o f acetonitrUe R and HjN N NH2
3 volumes o f water R.
Column: A. methyl 3,5-diamino-6-chloropyrazine-2-carboxylate.
— sizer. I = 0.25 m , 0 = 4.6 mm;
PhEur
— stationary phase: octadecylsHyl silica gel for chromatography R
(5 pm ).
Mobile phase M ix 5 volumes o f tetramethylammoraum hydroxide
solution R, 250 volumes o f acetonitrUe R and 745 volumes of
* * *★
water R ; adjust to p H 7.0 with a mixture o f 1 volume of Aminobenzoic Acid *
★ ★
phosphoric add R and 9 volumes o f water R. Adjust the
(4-Aminobenzoic Add, Ph Eur monograph 1687) * * * * *
concentration o f acetonitrile in the mobile phase so that the
retention time o f im purity A is 5-6 min (an increase in the
concentration o f acetonitrile results in a shorter retention
time). A djust die concentration o f tetramethylammonium
hydroxide and o f phosphoric a d d keeping the p H at 7.0 so
that the retention time of amiloride is 9-12 min (an increase
h 2n
XT
in the concentration results in a shorter retention time for
CzHyNOa 137.1 150-13-0
amiloride).
Flow rate 1 mL/min. A ctio n a n d u se
Detection Spectrophotom eter at 254 run. Skin protective.
Irqection 20 pL. PhEtr__________ __
Run time 5 times the retention time o f amiloride.
D E F IN IT IO N
System suitability: reference solution (b):
4-Aminobenzoic acid.
— signal-to-noise ratio: m inim u m 5.0 for the peak due to
amiloride. C o n te n t
Limits: 99.0 p er cent to 101.0 per cent (anhydrous substance).
— unspecified impurities: for each impurity, n o t more than CHARACTERS
0.2 tim es the area of the peak due to impurity A in the A p p e a ra n c e
chrom atogram obtained with reference solution (c) W hite o r slightly yellow, crystalline powder.
(0.10 per cent);
S o lu b ility
— total: not m ore than the area o f the peak due to
Slightly soluble in water, freely soluble in alcohol. It dissolves
im purity A in the chromatogram obtained with reference
in dilute solutions o f alkali hydroxides.
solution (c) (0.5 per cent);
— disregard limit. 0.1 times the area of the peak due to ID E N T IF IC A T IO N
im purity A in the chromatogram obtained with reference First identification B
solution (c) (0.05 per cent). Second identification A , C
Water (2.5.12) A. M elting point (2.2.14): 186 °C to 189 °C.
11.0 per cent to 13.0 per cent, determ ined on 0.200 g. B. Infrared absorption spectrophotometry (2.2.24).
Sulfated ash (2.4.14) Comparison 4-aminobenzoic add CRS.
M axim um 0.1 p er cent, determined on 1.0. g. C. Thin-layer chromatography (2.2.27).
1-140 Aminobenzoic Acid 2016
Test solution Dissolve 20 mg o f the substance to be examined — any other impurity: not more than 0.5 times the area o f the
in methanol R and dilute to 20 m L w ith the same solvent. peak due to im purity A in the chrom atogram obtained
Reference solution (a) Dissolve 20 m g o f 4-aminobenzoic with the reference solution (0.1 p er cent),
add CRS in methanol R and dilute to 20 m L with the same — total: n o t more than 2.5 times the area o f the peak due to
solvent. impurity A in the chrom atogram obtained with the
reference solution (0.5 p er cent),
Reference solution (b) Dissolve 10 m g o f 4-nitrobenzoic acid R
— disregard limit: 0.1 times the area o f the peak due to
in 10 m L o f reference solution (a).
impurity A in the chrom atogram obtained with the
Plate Suitable silica gel with a fluorescent indicator having an reference solution (0.02 per cent).
optimal intensity at 254 nm as the coatin g substance.
Impurity C and im purity D
Mobile phase glacial acetic add R, hexane R, methylene
Gas chromatography (2.2.28).
' chloride R (5:20:75 VIVIV).
Internal standard solution Dissolve 20.0 m g o f lauric add R in
Application 1 |iL.
methylene chloride R and dilute to 100.0 m L with the same
Development Over a path of 10 cm. solvent.
Drying In air. Test solution Dissolve 1.000 g o f the substance to be
Detection Examine in ultraviolet light at 254 nm. examined in 10.0 m L o f an 84 g/L solution o f sodium
System suitability T he chromatogram obtained with reference hydroxide R and extract with 2 quantities, each o f 10 mT, of
solution (b) shows 2 clearly separated spots. methylene chloride R. Combine and wash with 5 m L of
Results T he principal spot in the chrom atogram obtained with water R; filter through anhydrous sodium sulfate R. W ash the
the test solution is similar in position and size to the principal filter with methylene chloride R. Evaporate in a water-bath at
spot in the chromatogram obtained with reference 50-60 °C to obtain a volume o f about 1-5 mL. A dd 1.0 m L
solution (a). o f the internal standard solution and dilute to 10.0 m L with
methylene chloride R.
TESTS
Reference solution (a) Dissolve 20.0 m g o f aniline R in
A p p e a ra n c e o f so lu tio n
methylene chloride R and dilute to 100.0 m L w ith the same
The solution is clear (2.2.1) and no t m ore intensely coloured
solvent.
than reference solution B5 (2.2.2, Method II).
Reference solution (b) Dissolve 20.0 m g o f p-tohddine R in
Dissolve 1.0 g in alcohol R and dilute to 20 m L with the
methylene chloride R and dilute to 100.0 m L with the same
same solvent.
solvent.
Related substances Reference solution (c) Dilute 0.50 m L o f reference solution (a),
Liquid chromatography (2.2.29). 0.50 m L o f reference solution (b) and 10.0 m L o f the
Test solution Dissolve 25.0 mg o f the substance to be internal standard solution to 100.0 m L with methylene
examined in the mobile phase and dilute to 100.0 m L with chloride R.
the mobile phase. Column:
Reference solution Dissolve 25.0 m g o f 4-nitrobenzoic add R — material: fused silica,
and 25.0 m g o f benzocaine R in methanol R and dilute to — size. I = 30 m , 0 = 0.32 m m ,
100.0 m L with the same solvent. D ilute 1.0 m L to 50.0 m L — stationary phase. poly[methyl(95)phenyl(5)JsUoxane R (film
with the mobile phase. Dilute 1.0 m L o f this solution to thickness 0.5 nm).
10.0 m L with the mobile phase. Carrier gas helium for chromatography R.
Column: Flow rate 1.0 mL/min.
— size. I — 0.12 m , 0 = 4.0 m m ,
Split ratio 1:10.
— stationary phase: octylsUyl silica gel for chromatography R
(5 nm). Temperature:
Mobile phase Mix 20 volumes o f a mixture o f 70 volumes of
acetomtrUe R and 80 volumes o f methanol R, and 80 volumes H um Temperature
of a solution containing 1.5 g/L o f potassium dihydrogen (min) (°C)
Column 0 -4 130
phosphate R and 2.5 g/L of sodium octanesulfoncae R adjusted
to p H 2.2 with phosphoric add R. 4-6.5 130->180
Flow rate 1.0 mlVmin. 6.5 -11.5 180
Detection Spectrophotom eter at 270 nm . Injection port 280
Injection 20 |iL. 300
Detector
Run time 11 times the retention time o f 4-aminobenzoic add.
Relative retention W ith reference to 4-aminobenzoic a d d
(retention tim e = about 3 min): impurity A = about 4; Detection Flame ionisation.
impurity B = about 9. Injection 2 nL; inject the test solution and reference
Limits: solution (c).
— impurity A: not m ore than the area o f the corresponding Retention time Internal standard = about 9.5 min.
peak in the chromatogram obtained with the reference Limits:
solution (0.2 per cent), — impurity C: calculate th e ratio (K) o f the area o f the peak
— impurity B: n o t more than the area o f the corresponding due to impurity C to the area of the peak due to the
peak in the chromatogram obtained with the reference internal standard from the chromatogram obtained with
solution (0.2 per cent), reference solution (c); calculate the ratio o f the area o f the
peak due to impurity C to the area o f the peak due to the
internal standard from the chrom atogram obtained with
2016 Aminocaproic Acid 1-141
Identification of impurities Use the chromatogram obtained Prepare the reference solution vising lead standard solution
with reference solution (b) to identify die peak due to (1 ppm Pb) obtained by diluting lead standard solution
impurity A. (100 ppm Pb) R with a mixture o f 5 m L o f water R and
Relative retention W ith reference to aminoglutethimide 15 m L o f acetone R
(retention time = about 9 min): impurity A = about 1.3. L oss o n d ry in g (2.2.32)
System suitability Reference solution (d): M axim um 0.5 p er cent, determined on 1.000 g by drying in
— resolution: m inim u m 2.0 between the peaks due to an oven at 105 °C.
am in oglu teth im id e and impurity A. S u lfa te d a sh (2.4.14)
Limits: M axim um 0.1 per cent, determined on 1.0 g.
— impurity A: not m ore than twice die area o f the principal
A SSA Y
peak in the chromatogram obtained w ith reference
Dissolve 0.180 g in 50 m L of anhydrous acetic acid R and
solution (b) (2.0 p er cent);
titrate with 0.1 M perchloric acid, determining the end-point
— unspecified impurities: for each impurity, n o t more than
potentiometrically (2.2.20).
0.1 times the area o f the principal peak in die
chromatogram obtained with reference solution (c) 1 m L of 0.1 M perchloric acid is equivalent to 23.23 mg
(0.10 per cent); Of C 13H 16N 2O 2.
— sum of impurities other than A: not more than the area o f IM P U R IT IE S
the principal peak in die chromatogram obtained with Specified impurities A, D .
reference solution (c) (1.0 per cent);
Other detectable impurities (the following substances would, if
— total: maximum 2.0 per cent for the sum of die contents
present at a sufficient level, be detected by one o r other o f
o f all impurities;
the tests in the monograph. T hey are limited by the general
— disregard limit: 0.05 times the area of the principal peak in
acceptance criterion for other/unspecified impurities and/or
the chromatogram obtained with reference solution (c)
by the general monograph Substances for pharmaceutical use
(0.05 per cent).
(2034). It is therefore not necessary to identify these
Impurity D impurities for demonstration o f compliance. See also 5.10.
Liquid chromatography (2.2.29). Carry out die test protected Control of impurities in substances for pharmaceutical use): B, C.
from light Use shaking, not sordcadon or heat, to dissolve the
reference substance and the substance to be examined.
Test solution Dissolve 0.100 g of the substance to be
examined in dimethyl sulfoxide R and dilute to 100.0 m L with R4 and enantiomer
the same solvent.
Reference solution Dissolve 3.0 mg of aminoglutethimide
impurity D CRS in dimethyl sulfoxide R and dilute to
100.0 m L with the same solvent. Dilute 1.0 m L of this A. R3 = N H 2) R4 = H: (3RS)-3-(3-aminophenyl)-3-
solution to 100.0 m l, with dimethyl sulfoxide R. ethylpiperidine-2,6-dione (3-aminoglutethimide),
Column: B. R3 = N O * R4 = H:
— sizer. I = 0.12 m , 0 = 4 mm; (3i?S)-3-ethyl-3-(3-nitrophenyl)piperidine-2,6-dione,
— stationary phase: octadecylsUyl silica gel for chromatography R C. R3 = H , R4 = N 0 2:
(5 pm). (3i?5)-3-ethyl-3-(4-nitrophenyl)piperidine-2,6-dione,
Mobile phase Dissolve 0.285 g of sodium edetate R in water R,
add 7.5 m L o f dilute acetic acid R and 50 m l, of 0.1 M
potassium hydroxide and dilute to 1000 m L with water R;
adjust to p H 5.0 with glacial acetic acid R’, mix 350 m L of
this solution with 650 m L of methanol R.
Flow rate 1.0 mL/min.
Detection Spectrophotometer at 328 nm.
Injection 10 |iL.
System suitability T est solution: D . 3,3'-[diazenediylbis(4,l-phenylene)]bis(3-ethylpiperidine-
— number of theoretical plates: minimum 3300, calculated for 2, 6-dione) (azoglutethimide).
the principal peak; PhEur
— mass distribution ratio: 2.0 to 5.0 for the principal peak;
— symmetry factor, m axim um 1.2 for the principal peak.
Limit:
— impurity D: n o t m ore than the area of the principal peak
in the chromatogram obtained with the reference solution
(300 ppm).
Sulfates (2.4.13)
M aximum 500 ppm .
Dilute 6 m L o f solution S to 15 m L with distilled water R.
Heavy m etals (2.4.8)
M aximum 10 ppm.
Dissolve 2.0 g in 15 m L o f acetone R and dilute to 20 m L
with water R. 12 m L of the solution complies with test B.
1-144 Aminophylline 2016
A SSA Y
^ n^ v n
Etfaylenediamine
Dissolve 0.250 g in 30 m L of water R. Add 0.1 m L of
bromocresol green solution R. T itrate with 0.1 M hydrochloric 0
n>< 1 H
add until a green colour is obtained. ch3
0 pt
1 ' ★ ★
Aminophylline Hydrate
lx > 1
Çlheophyïïme-eàiylenediamine Hydrate,
★ ★
*****
ch3 Ph Eur monograph 0301)
A. 1,3,7-trimethyl-3,7-dihydro-lH-purine-2,6-dione
(caffeine),
, xH 20
H2N'
1 1 >
I
ch3
HN
X L //>
C 16H 24N 10O 4PCH2O 420.4 72487-55-9
CH,
(anhydrous substance)
B. 3-methyl-3 37-dîhydro-1H-purine-236-dione3
A c tio n a n d u se
Non-selective phosphodiesterase inhibitor; treatm ent of
A J. CHO
reversible airways obstruction.
O
JjC. N NH2
P re p a r a tio n
Aminophylline Injection
CH,
Aminophylline Tablets
Prolonged-release AminopyQine Tablets
C. AT-(6-am ino-l^-dim ethyl-2,4-dioxo-l,2,3,4-
tetrahydropyrimidin-5-yl)formamide, PhEur.
D E F IN IT IO N
C o n te n t
h3c x
— theophylline (C7H8N402; 180.2): 84.0 per cent to
»V>
hn ^ n
87.4 per cent (anhydrous substance);
— ethylenediamine (C 2H 8N 2; 60.1): 13.5 per cent to
CH, 15.0 per cent (anhydrous substance).
CHARACTERS
D. N-methyl-5-(methylamino)- lH-imidazole-4-carboxamide, A p p e a ra n c e
W hite or slightly yellowish powder, sometimes granular.
1-146 Aminophylline Hydrate 2016
1Y>
★ ★
H,c' N' l V ' N
(Ph. Eur. monograph 0803) * * * * *
o^ n- ^ n
ch3
H,C
A. 1,3,7-trim ethyl-3,7-dihydro-lH-purine-2,6-dione , HQ
(caffeine), N. .CH,
ÍJC> I
the same solvent.
Reference solution (a) Dissolve 10.0 m g of
ch3 (2-chloroethyl)diethylamine hydrochloride R (impurity H) in
methylene chloride R and dilute to .50.0 m L with the same
G . 3,7-dim ethyl-3,7-dihydro-lii-purine-2,6-dione solvent. Dilute 2.0 m L o f the solution to 20.0 m L with
(theobromine). methylene chloride R.
PhEur
Reference solution (b) Mix 2.0 m L o f the test solution and
2.0 m L o f reference solution (a).
Plate TLC silica gel F2 5 4 plate R.
1-148 Amiodarone Hydrochloride 2016
Mobile phase anhydrous formic add R, methanol R, methylene — disregard limit. 0.25 times the area o f the peak due to
chloride R (5:10:85 VIVIV). amiodarone in the chrom atogram obtained with the
Application 50 (iL o f the test solution and reference reference solution (0.05 per cent).
solution (a); 100 pL o f reference solution (b). Io d id e s
Development Over 2/3 o f the plate. M axim um 150 ppm.
Drying In a current o f cold air. Prepare the test and reference solutions simultaneously.
Detection Spray with potassium iodobismuthate solution R l and Solution A A dd 1.50 g o f the substance to be examined to
then with dilute hydrogen peroxide solution R', exam in e 40 m L o f water R at 80 °C and shake until completely
immediately in daylight. dissolved. Cool and dilute to 50.0 m L w ith water R.
System suitability: reference solution (b): Test solution T o 15.0 m L o f solution A add 1.0 m L o f
— the spot due to im purity H is clearly visible. 0.1 M hydrochloric acid and 1.0 mT. o f 0.05 Mpotassium
Limit: iodate. Dilute to 20.0 m L with water R. Allow to stand
— impurity H: any spot with the same Rp as the spot due to protected from light for 4 h.
im purity H in the chromatogram obtained with reference Reference solution T o 15.0 m L o f solution A add 1.0 m L of
solution (b) is n o t m ore intense th an the spot in the 0.1 M hydrochloric add, 1.0 m L of an 88.2 mg/L solution o f
chrom atogram obtained w ith reference solution (a) potassium iodide R and 1.0 m L o f 0.05 Mpotassium iodate.
(0.02 per cent). D ilute to 20.0 m L with water R. Allow to stand protected
Related substances from light for 4 h.
Liquid chromatography (2.2.29). M easure the absorbances (2.2.25) o f the solutions at 420 nm ,
Buffer solution pH 49 T o 800 mT. o f water R add 3.0 m L o f using a mixture o f 15.0 m L o f solution A and 1.0 m L o f
glacial acetic add R, adjust to p H 4.9 with dilute ammonia R l 0.1 M hydrochloric acid diluted to 20.0 m L with water R as
and dilute to 1000 m L with water R. the compensation liquid. T h e absorbance o f the test solution
is n o t greater than half the absorbance o f the reference
Test solution Dissolve 0.125 g o f the substance to be
solution.
ex a m in ed in a mixture of equal volumes of acetonitrUe R and
water R and dilute to 25.0 m L with the same mixture of H e a v y m e ta ls (2.4.8)
solvents. M axim um 20 ppm .
Reference solution Dissolve 5 m g o f amiodarone 1.0 g complies with test C. Prepare the reference solution
impurity D CRS, 5 m g of amiodarone impurity E CRS and using 2 m L o f lead standard solution (10 ppm Pb) R.
5.0 m g o f amiodarone hydrochloride CRS in methanol R and L o ss o n d ry in g (2.2.32)
dilute to 25.0 m L w ith the sam e solvent. Dilute 1.0 m L o f Maximum 0.5 per cent, determ ined on 1.000 g by drying at
the solution to 20.0 m L with a mixture o f equal volumes o f 50 °C at a pressure not exceeding 0.3 kPa for 4 h.
acetonitrile R and water R. S u lfa te d a s h (2.4.14)
Column: M aximum 0.1 per cent, determ ined on 1.0 g.
— size: I = 0.15 m , 0 = 4.6 mm;
— stationary phase, end-capped octadecylsUyl silica gel for A SSA Y
chromatography R (5 pm); Dissolve 0.600 g in a mixture o f 5.0 m L o f
— temperature: 30 °C. 0.01 M hydrochloric add and 75 m L o f ethanol (96 per cent) R.
Carry out a potentiom etric titration (2.2.20), using
Mobile phase Buffer solution p H 4.9, methanol R, acetonitrile R
0.1 M sodium hydroxide. Read the volume added between the
(30:30:40 VIVIV).
2 points o f inflexion.
Flow rate 1 mL/min.
1 mT. o f 0.1 M sodium hydroxide is equivalent to 68.18 m g of
Detection Spectrophotom eter at 240 nm . C 25H 3oCU2N 0 3 .
Injection 10 (iL.
STORAGE
Run time Twice the retention time o f am iodarone. Protected from light, at a tem perature n o t exceeding 30 °C.
Relative retention W ith reference to amiodarone (retention
time = about 24 m in): im purity A = about 0.26; IM P U R IT IE S
im purity D = about 0.29; im purity E = about 0.37; Specified impurities A, B, C , D , E, F , G, H
impurity B = about 0.49; im purity C = about 0.55;
im purity G = about 0.62; im purity F = about 0.69.
System suitability: reference solution:
— resolution: m inim um 3.5 between the peaks due to
impurities D and E.
Limits:
— impurities A , B, C, D, E, F, G: for each impurity, not
m ore than the area o f the peak due to amiodarone in the
A. (2-butyIbenzofuran-3-yl) [4-[2-(diethylamino)ethoxy]
chrom atogram obtained w ith the reference solution
phenyl] methanone,
(0.2 per cent);
— unspecified impurities: for each impurity, n o t more than
0.5 times the area o f the peak due to amiodarone in the
chrom atogram obtained w ith the reference solution
(0.10 per cent);
— total: not m ore th an 2.5 times the area of the peak due to
amiodarone in the chrom atogram obtained with the
reference solution (0.5 per cent);
2016 Amisulpride 1-149
and enantiomer ★* *
Amisulpride ★ ★
★ ★
(Ph Eur monograph 1490) *****
o\w/o
h 3c .
N
H u^
' ON and enantiomer.
B. (2-butylbenzofuran-3-yI) [4-[2-(ethyiamino)ethoxy]-3,5-
diiodophenyl] methanone, h2n och3 k CH3
and enantiomer
C 17H 27N 3O 4S 369.5 71675-85-9
DEFINITION
4-Amino-N- [ [(2i?5)-1-ethylpyrrolidin-2-yl] methyl] -5-
(ethylsulfonyl)-2-methoxybenzamide.
Content
99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
D . (2-butyIbenzofuran-3-yl)(4-hydroxy-3,5- Appearance
diiodophenyl)methanone, W hite or almost white, crystalline powder.
Solubility
H,c Practically insoluble in water, freely soluble in methylene
chloride, sparingly soluble in anhydrous ethanol.
mp
About 126 °C.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
E. (2-butyIbenzofuran-3-yl) (4-hydroxyphenyl)methanone,
Comparison amisulpride CRS.
TESTS
Appearance o f solution
T h e solution is clear (2.2.1) and not more intensely coloured
than reference solution Y6 (2.2.2, Method II).
Dissolve 1.0 g in 3 m L of a mixture of 1 volume of acetic
acid R and 4 volumes of water Rs and dilute to 20 m L with
F. (2-butylbenzofuran-3-yl) (4-hydroxy-3-iodophenyl) water R.
methanone, Impurity A
Thin-layer chromatography (2.2.27).
Test solution Dissolve 0.20 g of th e substance to be examined
in methanol R and dilute to 10 m L with the same solvent.
Reference solution (a) Dissolve 5 mg of sulpiride
impurity A CRS (amisulpride impurity A) in methanol R and
dilute to 25 mT. with the same solvent. Dilute 2 m L of the
solution to 20 m L with methanol R.
Reference solution (b) Dilute 1 m L of the test solution to
10 m L with reference solution (a).
G. [4-[2-(diethylamino)ethoxy]-3,5-diiodophenyl] [2-[(1 jR5)- Plate TLC silica gel G plate R.
1-methoxybutyl] benzofuran-3-yl] methanone,
Mobile phase 50 per cent VIV solution of concentrated
ammonia R, anhydrous ethanol R, di-isopropyl ether R
,c h 3
r
,N_XH3
(10:25:65 VIVIV); use the upper layer obtained after shaking
the mixture.
Application 10 (iL.
H . 2-chloro-N,N-diethylethanamine (2-chlorotriethylamine, Development Over 2/3 of the plate.
(2-chloroethyl)diethyIamine). Drying In air.
1-150 Amisulpride 2016
Detection Spray with ninhydrin solution R and heat at — reporting threshold: 0.05 per cent.
100-105 °C for 15 min. H ea v y m e ta ls (2.4.8)
Retardation factors Im purity A = about 0.2; M axim um 10 ppm.
amisulpride = about 0.5. Dissolve 4.0 g by gently heating in 5 m L o f dilute acetic
System suitability T he chrom atogram obtained with reference add R. Allow to cool and dilute to 20 m L w ith water R.
solution (b) shows 2 clearly separated spots. 12 m L o f the solution complies with test A. Prepare the
Limit. reference solution using lead standard solution (2 ppm Pb) R.
— impurity A: any spot due to impurity A is not more L o ss o n d ry in g (2.2.32)
intense than the corresponding spot in the chromatogram Maximum 0.5 per cent, determ ined on 1.000 g by drying in
obtained with reference solution (a) (0.1 per cent). an oven at 105 °C for 3 h.
R e la te d su b s ta n c e s S u lfa te d a s h (2.4.14)
liq u id chromatography (2.2.29). M axim um 0.1 per cent, determ ined on 1.0 g.
Solvent mixture acetomtrUe R l, methanol R2, mobile phase A A SSA Y
(12:16:72 V/V/V).
Dissolve 0.300 g with shaking in a mixture o f 5 m L o f acetic
Test solution Dissolve 0.10 g o f the substance to be examined anhydride R and 50 m L o f anhydrous acetic acid R. T itrate
in 16 m L o f methanol R2, add 12 m L of acetomtrUe R l and w ith 0.1 M perchloric add, determining the end-point
dilute to 100.0 m L with mobile phase A. potentiometrically (2.2.20).
Reference solution (a) Dilute 1.0 m L o f the test solution to 1 m L o f 0.1 M perchloric add is equivalent to 36.95 m g o f
100.0 m L with the solvent mixture. Dilute 1.0 m L o f this C 17H 2 7N 3O 4 S .
solution to 10.0 m L with the solvent mixture.
IM P U R IT IE S
Reference solution (b) Dissolve the contents o f a vial of
Spedfied impurities A
amisulpride for system suitabUity CRS (containing impurity B)
in 1.0 m L of the solvent mixture. Other detectable impurities (the following substances w ould, if
present at a sufficient level, be detected by one or other o f
Column:
the tests in the monograph. They are lim ited by th e general
— size: I = 0.25 m , 0 = 4.6 mm ;
acceptance criterion for other/unspecified impurities and/or
— stationary phase:, base-deactivated oaylsüyl sUica gel for
by the general monograph Substances for pharmaceutical use
chromatography R (5 nm);
— temperature: 40 °C.
(2034). It is therefore not necessary to identify these
impurities for demonstration o f compliance. See also 5.10.
Mobile phase:
Control of impurities in substances for pharmaceutical use): B, C,
— mobile phase A: dissolve 0.7 g o f sodium octanesulfonate R
D, E, F, G3 H.
in 930 m L o f water R , add 45.0 m l. o f a 5 per cent V/V
solution o f dilute sulfuric acid R, adjust to p H 2.3 with a
5 per cent V/V solution o f dilute sulfuric add R, and dilute h2n
to 1000 m L with water R; H N and enantiomer
— mobile phase B: methanol R2;
CH3
— mobile phase C: acetomtrUe R l :;
18 - 35 72 50 16 38 12
\
and enantiomer
o o Content
*o
h 3c N -^ O 99.0 per cent to 101.0 per cent (dried substance).
and enantiomer
H H N
h2n och3
CHARACTERS
CH3 Appearance
W hite or almost white powder or colourless crystals.
D . 4-am ino-N - [ [ (2RS)-1 -ethylpyrrolidin-2-yl] methyl] -2- Solubility
methoxy-5-(methylsulfonyl)benzamide, Freely soluble in water, in ethanol (96 per cent) and in
methylene chloride.
o o
IDENTIFICATION
H2N.XX. ^ OCH3
A. Infrared absorption spectrophotometry (2.2.24).
Comparison amitriptyline hydrochloride CRS.
B. 20 m g gives reaction (a) of chlorides (2.3.1).
E. 4-amino-5-(ethylsulfonyl)-2-methoxybenzoic acid.
TESTS
Appearance of solution
T he solution is clear (2.2.1) and not more intensely coloured
na QN
H and enantiomer than reference solution B7 (2.2.2, Method II).
I '0
h2n 0CH3 I Dissolve 1.25 g in water R and dilute to 25 m L with the
ch3
same solvent
h3c n^ ^ o ^ s^ nh2
SOjH
,0 . Jl 1 .0 , .c h 3
H
*c Yo t
:
CT
o
^
A. 10,11 -dihydro-5H-dibenzo [a,d\ [7] annulen-5-one
(dibenzosuberone), o ci and enantiomer
PhEur--------------------------------------------------------------------------------
B. 3-(5if-dibenzo[a,d] [7]annulen-5-ylidene)-isyV-
D E F IN IT IO N
dimethylpropan-1-amine (cydobenzaprine),
3-Ethyl 5-methyl (4R^)-2-[(2-aminoethoxy)methyl]-4-
(2-chlorophenyl)-6-methyl-l,4-dihydropyridine-3,5-
dicarboxylate benzenesulfonate.
.C H ,
C o n te n t
97.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
A p p e a ra n c e
C. 3-(10,l l-dihydro-5H-dibenzo[a,d] [7]annulen-5-ylidene)- W hite or almost white powder.
N -m ethylpropan-1-am ine (nortriptyline),
2016 Amlodipine Besilate 1-153
h3c x
NH
a
and enantiomer
DEFINITION
Content
25.0 per cent m/m to 30.0 p er cent mtm.
CHARACTERS
Appearance
Clear, colourless liquid, very caustic.
Solubility
Miscible with water and with ethanol (96 per cent).
Carbonates ★ ★
M aximum 60 ppm.
Ammonio Methacrylate Copolymer ★ ★
T o 10 m L in a test-tube with a ground-glass neck add (Type A) *****
10 m l, o f calcium hydroxide solution R. Stopper immediately (Ph Eur monograph 2081)
and mix. Any opalescence in the solution is not more intense
than that in a standard prepared at the same time and in the
r
° 1
0
same manner using 10 m L o f a 0.1 g/L solution of anhydrous
J k 0 / * \ ch3
O
X
H2C ^
«0
\
sodium carbonate R.
O
2
-
X
C h lo rid e s (2.4.4) ch3
M aximum 1 ppm.
Dilute 5 m L o f solution S to 15 m L with water R.
S u lfates (2.4.13)
cr
Maximum 5 ppm.
Dilute 3 m L o f solution S to 15 m L with distilled water R.
Iro n (2.4.9)
A ctio n a n d u se
M aximum 0.25 ppm.
Excipient.
Dilute 4 m L o f solution S to 10 m L with water R.
H eavy m e ta ls (2.4.8) PhEir.
M aximum 1 ppm. D E F IN IT IO N
Dilute 4 m L o f solution S to 20 m L with water R. 12 m L o f Poly(ethyl propenoate-co-methyl 2-methylpropenoate-co-2-
the solution complies with test A. Prepare the reference (trimethylammonio)ethyl 2-methylpropenoate) chloride
solution using lead standard solution (2 ppm Pb) R. having a m ean relative molecular mass of about 150 000.
R esid u e o n e v a p o ra tio n T h e ratio o f ethyl propenoate groups to methyl
Maximum 20 m g/L 2-methylpropenoate groups to 2-(trimethylammonio)ethyl
Evaporate 50 m L to dryness on a water-bath and dry at 2-methylpropenoate groups is about 1:2:0.2.
100-105 °C for 1 h. T he residue weighs a maximum of C o n te n t o f a m m o n io m e th a c ry la te g ro u p s
1 mg. 8.9 per cent to 12.3 per cent (dried substance).
ASSAY CHARACTERS
Weigh accurately a flask with a ground-glass neck containing A p p e a ra n c e
50.0 m L o f 1 M hydrochloric add. A dd 2 m L of the substance Colourless to white or almost white granules or powder.
to be examined and re-weigh. Add 0.1 m L of methyl red S o lu b ility
solution R as indicator. T itrate with 1 M sodium hydroxide Practically insoluble in water, freely soluble in anhydrous
until the colour changes from red to yellow. ethanol and in methylene chloride giving clear to cloudy
1 m L of 1 M hydrochloric acid is equivalent to 17.03 m g of solutions. D ue to the polymeric nature o f the substance, a
N H 3. stirring time of u p to 5 h may be necessary.
STORAGE ID E N T IF IC A T IO N
Protected from air, at a temperature not exceeding 20 °C. A. Infrared absorption spectrophotometry (2.2.24).
__________________________________________________________ PhEur Comparison Ph Eur. reference spectrum of ammonio methacrylate
copolymer (type A ).
B. Viscosity (see Tests).
C. It complies with the limits o f the assay.
TESTS
S o lu tio n S
Dissolve a quantity of the substance to be examined
corresponding to 12.5 g of the dried substance in a mixture
o f 35.0 g o f acetone R and 52.5 g of 2-propanol R.
V iscosity (2.2.10)
M aximum 15 mPa-s, determined on solution S.
Apparatus Rotating viscometer.
Dimensionr.
— spindle: diameter = 25.15 mm ; height = 90.74 mm ; shaft
diam eter = 4.0 mm;
— ctfinder. diameter = 27.62 mm ; height = 0.135 m .
Stirring speed 30 r/min.
Volume of solution 16 m L of solution S.
Temperature 20 °C.
A p p e a ra n c e o f a film
Spread 2 m L of solution S evenly on a glass plate. U pon
drying a clear film is formed.
1-156 Ammonio Methacrylate Copolymer 2016
M o n o m e rs
L iquid chrom atography (2.2.29).
Ammonio Methacrylate Copolymer * * * * *
★ ★
Solution A Dissolve 3.5 g o f sodium perchlorate R in water for (Type B) * * * * *
chromatograpky R and dilute to 100 m L with the same (Ph Eur monograph 2082)
solvent.
Test solution Dissolve 5.00 g of the substance to be examined o
in methanol R and dilute to 50.0 m L with the same solvent.
HjC ^ A ^ C H s
T o 10.0 m L o f this solution add 5.0 m L o f solution A, o ch3
dropwise, while continuously stirring. Remove the ch3
precipitated polymer by centrifugation. U se the clear
supernatant solution. 9 H,C CH3
Reference solution Dissolve 50.0 m g o f ethyl acrylate R and H*0 -"•'C H , cr
10.0 m g o f methyl methacrylate R in methanol R and dilute to
50.0 m L with the sam e solvent. Dilute 1.0 m L o f the
solution to 100.0 m L with methanol R. A dd 10 m L o f this
solution to 5 m L of solution A.
Column: A c tio n a n d u se
— sizer. I = 0.12 m , 0 = 4.6 mm; Excipient
— stationary phase: octadecylsUyl silica gel for chromatography R PhEur_____________
(7 Jim).
D E F IN IT IO N
Mobile phase Dilute phosphoric add R with water for
chromatography R to obtain a solution at p H 2.0; Poly(ethyl propenoate-co-m ethyl 2-m ethylpropenoate-co-2-
(trimethylammpnio)ethyl 2-m ethylpropenoate) chloride
mix 800 m L o f this solution and 200 m L o f methanol R, filter
and degas. having a m ean relative m olecular mass o f about 150 000.
Flow rate 2.0 mL/min. T h e ratio o f ethyl propenoate groups to methyl
Detection Spectrophotom eter at 202 nm . 2-methylpropenoate groups to 2-(trimethylam monio)ethyl
2-methylpropenoate groups is about 1:2 :0 . 1.
Injection 50 pL.
System suitability: reference solution: C o n te n t o f a m m o n io m e th a c ry la te g ro u p s
— resolution: minimum 1.5 between the peaks due to 4.5 per cent to 7.0 per cent (dried substance).
impurity A and im purity B. CHARACTERS
Limits: A p p e a ra n c e
— impurity A: not m ore than the area of the corresponding Colourless to white or almost white granules or powder.
peak in the chrom atogram obtained w ith the reference
S o lu b ility
solution (100 ppm );
Practically insoluble in water, freely soluble in anhydrous
— impurity B: not m ore than 2.5 times the area of the ethanol and in methylene chloride giving clear to cloudy
corresponding peak in the chrom atogram obtained with solutions. D ue to the polymeric nature o f die substance, a
the reference solution (50 ppm). stirring time o f up to 5 h may be necessary.
M e th a n o l (2.4.24, System A)
M axim um 1.5 per c e n t ID E N T IF IC A T IO N
A. Infrared absorption spectrophotom etry (2.2.24).
H e a v y m e ta ls (2.4.8)
M axim um 20 ppm. Comparison Ph. Eur. reference spectrum of ammonio methacrylate
1.0 g complies with test C. Prepare the reference solution copolymer (type B).
using 2.0 m L of lead standard solution (10 ppm Pb) R. B. Viscosity (see Tests).
L o ss o n d ry in g (2.2.32) C. It complies w ith the limits of the assay.
M axim um 3.0 per cent, determ ined on 1.000 g by drying TESTS
in vacuo at 80 °C for 5 h. S o lu tio n S
A SSA Y Dissolve a quantity o f die substance to be examined
Dissolve 1.000 g in a mixture o f 3 mL o f anhydrous formic corresponding to 12.5 g o f the dried substance in a m ixture
add R and 30 m L o f anhydrous acetic add R and heat to of 35.0 g of acetone R and 52.5 g o f 2-propanol R.
dissolve. A dd 20 m L o f acetic anhydride R. T itrate with 0.1 M V iscosity (2.2.10)
perchloric acid, determining the end-point potentiometrically M axim um 15 mPa-s, determ ined on solution S.
(2.2.20).
Apparatus Rotating viscometer.
1 m L of 0.1 M perchloric add is equivalent to 20.77 mg
o f C 9H 1802 N C 1 (ammonio methacrylate groups).
Dimensions:
— spindle: diam eter = 25.15 m m ; height = 90.74 m m ; shaft
IM P U R IT IE S diam eter = 4.0 mm;
Specified impurities A, B — cylinder, diam eter = 27.62 m m ; height = 0.135 m.
Stirring speed 30 r/min.
HjC Volume of sdution 16 m L of solution S.
Temperature 20 °C.
A. R = H , R ; = C 2H 5: ethyl propenoate (ethyl acrylate), A p p e a ra n c e o f a film
Spread 2 m L o f solution S evenly on a glass plate. U p o n
B. R = R ' = C H 3: methyl 2-m ethyipropenoate (methyl
drying a clear film is formed.
methacrylate).
PhEur
2016 Ammonium Bicarbonate 1-157
M o n o m e rs
L iquid chromatography (2.2.29).
Ammonium Bicarbonate ?**%
Solution A Dissolve 3.5 g of sodium perchlorate R in water for *★ ★*
(Ammonium Hydrogen Carbonate, *
chromatography R and dilute to 100 m L with the same Ph Eur monograph 1390)
solvent. NHtHCOa 79.1 1066-33-7
Test solution Dissolve 5.00 g of the substance to be examined
in methanol R and dilute to 50.0 m L with the same solvent. A ctio n a n d u se
T o 10.0 m L of this solution add 5.0 m L o f solution A, Expectorant
dropwise, while continuously stirring. Remove the P re p a ra tio n s
precipitated polymer by centrifugation. Use the clear Aromatic Ammonia Solution
supernatant solution.
Strong Ammonium Acetate Solution
Reference solution Dissolve 50.0 mg of ethyl acrylate R and
10.0 m g o f methyl methacrylate R in methanol R and dilute to Aromatic Ammonia Spirit
50.0 m L with the same solvent. Dilute 1.0 m L of the PhEur__________________________________________________________
solution to 100.0 m L with methanol R. Add 10 m L o f this
solution to 5 m L o f solution A. D E F IN IT IO N
Column: C o n ten t
— sizer. I = 0.12 m , 0 = 4.6 mm; 98.0 per cent to 101.0 per cent.
— stationary phase: octadecylsSyl silica gel for chromatography R CHA RACTERS
(7 pm). A p p e a ra n c e
Mobile phase Dilute phosphoric acid R with water for Fine, white or almost white, crystalline powder or white or
chromatography R to obtain a solution at p H 2.0; almost white crystals, slightly hygroscopic.
mix 800 m L of this solution and 200 m L o f methanol R, filter Solubility
and degas. Freely soluble in water, practically insoluble in ethanol
Flow rate 2.0 mL/min. (96 per cent).
Detection Spectrophotometer at 202 nm. It volatilises rapidly at 60 °C. T h e volatilisation takes place
Injection 50 (iL. slowly at ambient temperatures if the substance is slightly
System suitability: reference solution: moist. It is in a state of equilibrium with ammonium
— resolution: minimum 1.5 between the peaks due to carbamate.
impurity A and impurity B.
ID E N T IF IC A T IO N
Limits: A. It gives the reaction o f carbonates and bicarbonates
— impurity A: n o t more than the area o f the corresponding
(2.3.1).
peak in the chromatogram obtained with the reference
B. Dissolve 50 m g in 2 m L of water R. T he solution gives the
solution (100 ppm);
— impurity B: not more than 2.5 times the area o f the reaction o f amm onium salts (2.3.1).
corresponding peak in the chromatogram obtained with TESTS
the reference solution (50 ppm). S o lu tio n S .
M e th a n o l (2.4.24, System A) Dissolve 14.0 g in 100 m L of distilled water R. Boil, to remove
M aximum 1.5 per c e n t the ammonia, allow to cool and dilute to 100.0 m L with
H eav y m e ta ls (2.4.8) distilled water R.
M axim um 20 ppm. C h lo rid es (2.4.4)
1.0 g complies with test C. Prepare the reference solution Maximum 70 ppm .
using 2.0 m L of lead standard solution (10 ppm Pb) R. Dilute 5 m L of solution S to 15 m L with water R.
L oss o n d ry in g (2.2.32) S u lfates (2.4.IS)
M axim um 3.0 p er cent, determined on 1.000 g by drying Maximum 70 ppm , determined on solution S.
in vacuo at 80 °C for 5 h.
Iro n (2.4.9)
A SSAY Maximum 40 ppm .
Dissolve 2.000 g in a mixture of 3 m L of anhydrous formic
Dilute 1.8'm L o f solution S to 10 m L with water R.
acid R and 30 m L o f anhydrous acetic add R and h eat to
dissolve. Add 20 m L o f acetic anhydride R. Titrate with 0.1 M H eav y m e ta ls (2.4.8)
perchloric add, determining the end-point potentiometrically Maximum 10 ppm .
( 2.2.20). Dissolve cautiously 2.5 g in 25 m L o f 1 M hydrochloric acid.
1 m L of 0.1 M perchloric add is equivalent to 20.77 mg 12 m L of the solution complies with test A. Prepare the
o f C 9H i80 2N C 1 (ammonio methacrylate groups). reference solution using lead standard solution (1 ppm Pb) R.
IM P U R IT IE S ASSAY
Specified impurities A, B Dissolve cautiously 1.0 g in 20.0 m L o f 0.5 M sulfuric acid
o and dilute to 50 m L with water R. Boil, cool and titrate the
excess of a d d w ith 1 M sodium hydroxide, using 0.1 m L o f
HiCY ^ 0' R methyl red solution R as indicator.
R 1 m L o f 0.5 M sulfuric acid is equivalent to 79.1 mg
A. R = H , R ; = C 2H 5: ethyl propenoate (ethyl acrylate), o fN H tH C O s.
B. R = R ' = C H 3: methyl 2-methylpropenoate (methyl ST O R A G E
methacrylate). In an airtight container.
________________________________ !___________ ;______________ PhEur _________________________________________________________ PhEur
1-158 Ammonium Bromide 2016
TESTS ***
Amoxicillin Sodium ★ *
S o lu tio n S ★ ★
Dissolve 5.0 g in alcohol (50 per cent V/V) R and dilute to (Ph. Eur. monograph 0577) *****
50 m L with the same solvent.
A p p e a ra n c e o f so lu tio n ! COzHa
Solution S is clear (2.2.1) and not m ore intensely coloured
than reference solution Y7 (2.2.2, Method II).
p H (2.2.3)
Dissolve 5.0 g in carbon dioxide-free water R and dilute to
50 m L with the same solvent. Disregard any slight residue.
T h e p H of the solution is n o t m ore than 11.0.
C 16H18N 3N a 0 5S 387.4 34642-77-8
R e la te d su b s ta n c e s
Examine by thin-layer chromatography (2.2.27), using silica A c tio n a n d u se
gel GF254 R as the coating substance. Penicillin antibacterial.
Test solution Dissolve 1.0 g o f the substance to be examined P re p a r a tio n s
in alcohol R and dilute to 100 m L w ith the same solvent. Amoxicillin Injection
Reference solution D ilute 0.5 m L of the test solution to Co-amoxiclav Injection
100 m L with alcohol R.
PhEur.
Apply separately to the plate 20 jiL o f each solution. Develop
over a path o f 15 cm using the lower layer of a mixture of D E F IN IT IO N
5 volumes o f concentrated ammonia R, 15 volumes o f alcohol R Sodium (2S,5R,6R)-6-[[(2i?)-2-amino-2-(4-hydroxyphenyl)
and 80 volumes of chloroform R. Examine the plate acetyl] amino] -3,3-dimethyl-7 -oxo-4-thia-1 -
immediately in ultraviolet light at 254 nm . Spray with azabicydo[3.2.0]heptane-2-carboxylate.
diphenylcarbazone mercuric reagent R. Allow the plate to dry in Semi-synthetic product derived from a fermentation product.
air and spray with freshly prepared alcoholic potassium
C o n te n t
hydroxide, solution R diluted 1 in 5 with aldehyde-free alcohol R.
89.0 per cent to 102.0 per cent (anhydrous substance).
H eat at 100 °C to 105 °C for 5 m in and exam in e
immediately. W hen examined in ultraviolet light and after CHARACTERS
spraying, any spot in the chromatogram obtained with the A p p e a ra n c e
test solution, apart from the principal spot, is no t more W hite or almost white, very hygroscopic, powder.
intense than the spot in the chromatogram obtained with the S o lu b ility
reference solution (0.5 per cent). Disregard any spot at the Very soluble in water, sparingly soluble in anhydrous ethanol,
point o f application. very slightly soluble in acetone.
L oss o n d ry in g (2.2.32) ID E N T IF IC A T IO N
N o t m ore than 3.0 per cent, determined on 0.50 g by drying
First identification A , D.
in an oven at 130 °C.
Second identification B, C, D.
A SSA Y
A. Infrared absorption spectrophotom etry (2.2.24).
Dissolve 0.200 g in 5 m L o f ethanol R. Add 0.5 m L o f
thymolphthalein solution R and 10 m L o f silver nitrate solution Preparation Dissolve 0.250 g in 5 m L o f water R, add 0.5 m L
o f dilute acetic add R, swirl and allow to stand for 10 m in in
in pyridine R. T itrate with 0.1 M ethanolic sodium hydroxide
iced water. Filter die crystals and wash with 2-3 m L o f a
until a pure blue colour is obtained. Carry out a blank
titration. mixture o f 1 volume o f water R and 9 volumes o f acetone R,
then dry in an oven at 60 °C for 30 min.
1 m L oi 0.1 M ethanolic sodium hydroxide is equivalent to
24.83 m g o f C n H jy ^ N a C ^ .
Comparison amoxicillin trihydrate CRS.
B. Thin-layer chromatography (2.2.27).
STORAGE
Test solution Dissolve 25 m g o f die substance to be examined
Store in an airtight container.
in 10 m L o f sodium hydrogen carbonate solution R.
PhEur
Reference solution (a) Dissolve 25 m g o f amoxicillin
trihydrate CRS in 10 m L o f sodium hydrogen carbonate
solution R.
Reference solution (b) Dissolve 25 m g o f amoxicillin
trihydrate CRS and 25 mg o f ampidHin trihydrate CRS in
10 m L o f sodium hydrogen carbonate solution R.
Plate TLC sUamsed silica gel plate R.
Mobile phase M ix 10 volumes o f acetone R and 90 volumes of
a 154 g/L solution o f ammonium acetate R previously adjusted
to p H 5.0 with glacial acetic acid R.
Application 1 |jL.
Development Over a path o f 15 cm.
Drying In air.
Detection Expose to iodine vapour until the spots appear and
examine in daylight-
2016 Amoxicillin Sodium 1-163
System suitability: reference solution (b): — mobile phase B: mix 20 volumes of acetonitrüe R and
— the chromatogram shows 2 clearly separated spots. 80 volumes o f a 25 per cent V/V solution o f 0.2 M
Results T h e principal spot in the chromatogram obtained with potassium dihydrogen phosphate R adjusted to p H 5.0 with
the test solution is similar in position, colour and size to the dilute sodium hydroxide solution R\
principal spot in the chromatogram obtained with reference
solution (a). Time Mobile phase A Mobile phase B
C. Place about 2 m g in a test-tube about 150 m m long and (min) (per cent V/V) (per cent V/V)
about 15 mm in diameter. M oisten with 0.05 m L o f water R 0 -t, 92 8
and add 2 m L o f sulfuric acid-formaldehyde reagent R. M ix the f ,- ( f , + 25) 92 -» 0 8 4 100
contents o f the tube by swirling; the solution is practically
colourless. Place the test-tube in a water-bath for 1 min; (Í, + 25) - (Í, + 40) 0 100
a dark yellow colour develops. (t^ + íO M ^ + ss) 92 8
D . It gives reaction (a) o f sodium (2.3.1). t, = retention time of amoxicillin determined with reference solution (c)
TESTS
A p p e a ra n c e o f so lu tio n I f the mobile phase has been adjusted to achieve the required
T h e solution is n o t more opalescent than reference resolution, the adjusted composition will apply at time zero
suspension II (2.2. 1), it may show an initial, b u t transient, in the gradient and in the assay.
pink colour, and after 5 min, its absorbance (2.2.25) at
Flow rate 1.0 mL/min.
430 nm is not greater than 0.20.
Detection Spectrophotom eter at 254 nm.
Dissolve 1.0 g in water R and dilute to 10.0 m L with the
same solvent Examine immediately after dissolution.
Injection 50 |iL o f reference solutions (b) and (c) with
isocratic elution at the initial mobile phase composition and
p H (2.2.3) 50 |iL o f test solution (b) and reference solution (d)
8.0 to 10.0. according to the elution gradient described under Mobile
Dissolve 2.0 g in carbon dioxide-free water R and dilute to phase; inject mobile phase A as a blank according to the
20 m L with the same solvent. elution gradient described under Mobile phase.
S p ecific o p tic a l ro ta tio n (2.2.7) Identification of impurities Use the chromatogram obtained
+ 240 to + 290 (anhydrous substance). w ith reference solution (d) to identify the 3 principal peaks
Dissolve 62.5 m g in a 4 g/L solution of potassium hydrogen eluted after the m ain peak corresponding to impurity C,
phthalate R and dilute to 25.0 m L with the same solution. amoxicillin dim er (impurity J; n = 1) and amoxicillin trim er
(impurity J; n = 2).
R e la te d su b sta n c e s
Liquid chromatography (2.2.29). Relative retention W ith reference to amoxicillin:
im purity C = about 3.4; impurity J (n = 1) = about 4.1;
Test solution (a) Dissolve 30.0 mg o f the substance to be
impurity J (n = 2) = about 4.5.
examined in mobile phase A and dilute to 50.0 m L with
mobile phase A. System suitability, reference solution (b):
— resolution: m inim um 2.0 between the peaks due to
Test solution (b) Dissolve 30.0 m g o f the substance to be
amoxicillin and cefadroxil; if necessary, adjust the ratio
examined in mobile phase A and dilute to 20.0 m L with
A:B o f the mobile phase.
mobile phase A. Prepare immediately before use.
Limits:
Reference solution (a) Dissolve 30.0 m g o f amoxicillin — impurity J (n = 1): not more than 3 times the area o f the
trihydrate CRS in mobile phase A and dilute to 50.0 m L with principal peak in the chromatogram obtained with
mobile phase A. reference solution (c) (3 per cent);
Reference solution (b) Dissolve 4.0 m g of cefadroxü CRS in — any other impurity: for each impurity, n o t more than twice
mobile phase A and dilute to 50 m L with mobile phase A. the area o f th e principal peak in the chromatogram
T o 5.0 m L o f this solution add 5.0 m L of reference obtained w ith reference solution (c) (2 per cent);
solution (a) and dilute to 100 m L with mobile phase A. — total: not m ore than 9 times the area o f the principal peak
Reference solution (c) Dilute 2.0 m L o f reference solution (a) in the chromatogram obtained with reference solution (c)
to 20.0 m L with mobile phase A. Dilute 5.0 m L of this (9 per cent);
solution to 20.0 m L with mobile phase A. — disregard limit. 0.1 times the area of the principal peak in
Reference solution (d) T o 0.20 g of amoxicillin trihydrate R add the chromatogram obtained w ith reference solution (c)
1.0 m L o f water R. Shake and add dropwise dilute sodium (0.1 per cent).
hydroxide solution R to obtain a solution. T he p H of the iV ^V -D im ethylaniline (2.4.26, Method A or B)
solution is about 8.5. Store the solution at room tem perature M aximum 20 ppm.
for 4 h. Dilute 0.5 m L of this solution to 50.0 m L with 2 -E th y lh ex a n o ic a c id (2.4.28)
mobile phase A. Maximum 0.8 per cent m/m.
Column:
H e a v y m e ta ls (2.4.8)
— sizer. I = 0.25 m , 0 = 4.6 mm;
M aximum 20 ppm.
— stationary phase: octadecylsüyl silica gel for chromatography R
(5 pm ). 1.0 g complies with test C. Prepare the reference solution
using 2 m L o f lead standard solution (10 ppm Pb) R.
Mobile phase:
— mobile phase A: mix 1 volume o f acetonitrüe R and W a te r (2.5.12)
99 volumes o f a 25 per cent V/V solution o f 0.2 M M axim um 3.0 p er cent, determined on 0.400 g.
potassium dihydrogen phosphate R adjusted to p H 5.0 with
dilute sodium hydroxide solution R]
1-164 Amoxicillin Sodium 2016
o V ^ H
V nA ^ h,
H2n - -|— k -S CHa
H H
G. (2S,5R,6R)-6 - [[(2R)-2-[[(2Æ)-2-amino-2-
A. (2S,5/?,6i?)-6-amino-3,3-dimethyl-7-oxo-4-thia-l- (4-hydroxyphenyl) acetyl] amino] 2-
azabicyclo[3.2.0]heptane-2-carboxylic a d d (4-hydroxyphenyl) acetyl] amino]-3,3-dim ethyl-7-oxo-4-thia-1-
(6-aminopenidllanic acid), azabicydo[3.2.0]heptane2-carboxylic a d d
(D-(4-hydroxyphenyl)glycylamoxidllm),
h3c 0
HjCJ__ff
h-î / h !NH
C02H
B. (2S,52?,6Æ)-6-[[(2S)-2-amino-2-(4-hydroxyphenyl)acetyl]
amino] -3,3-dime thyl-7-oxo-4-thia-1- H . (2i?)-2-[(2,2-dimethylpropanoyl)amino]-2-
azabicyclo[3.2.0]heptane-2-caiboxylic acid (L-amoxicillin), (4-hydroxyphenyl) acetic ad d ,
*1 COzH
H NH2 u H N -A .C H 3
CO2H
D . (45)-2-[ [[(2Æ)-2-amino-2-(4-hydroxyphenyl)acetyI]
amino] carboxymethyl]-5,5-dimethylthiazolidine-4-carboxylic
J. co-oligomers of amoxidllin and penidlloic ad d s of
a d d (penidlloic ad d s o f amoxicillin),
amoxicillin,
2016 Amoxicillin Trihydrate 1-165
O > C°2H
V i ><CH3 G . (2S,5R,6R)-6-[ [(2R)-2-[[(2J?)-2-amino-2-
CHa (4-hydroxyphenyI) acetyl] amino]-2-(4-hydroxy-
H H
phenyl)acetyl]amino]-3,3-dimethyi-7-oxo-4-thia-l-
azabicydo[3.2.0]heptane-2-carboxylic a d d
A. (25,5J?,6i?)-6-ammo-3,3-dimethyl-7-oxo-4-thia-l- (D-(4-hydroxyphenyl)glycylamoxidllin),
azabicydo [3.2.0]heptane-2-carboxylic a d d
(6-aminopenidllanic a d d ),
2016 Amphotericin 1-167
CH3 o
H3 Amphotericin * ★
★ ★
H3C H NH
(Amphotericin B, Ph Eur monograph 1292)
C 02H
C 4 7H 73 N O 17 924 1397-89-3
PhEur______________________
D E F IN IT IO N
Mixture o f antifungal polyenes produced by the growth o f
certain strains o f Streptomyces nodosus or obtained by any
other means. It consists mainly o f amphotericin B which is
J. co-oligomers of amoxicillin and o f penidlloic adds of (1R,3S,5R,6R,9R,UR,15S,16R,11R,18S,19E,21E,23E,~
amoxicillin, 25E,21E,29E,31E,33R,35S,36R,31S)-33-[(3-aiDmo-3,6-
dideoxy-P-D-mannopyranosyl)oxy]-13j5,6,9,ll,17,37-
octahydroxy-15,16,18-trimethyl-13-oxo-14,3 9-dioxa-
bicyclo [33.3.1] nonatriaconta-19,21,23,25,27,29,31-heptaene-
36-caiboxylic ad d .
Content
M inim um 750 IU /m g (dried substance).
CHARACTERS
Appearance
Yellow or orange, hygroscopic powder.
K . oligomers o f penidlloic adds o f amoxicillin, Solubility
Practically insoluble in water, soluble in dimethyl sulfoxide
and in propylene glycol, slightly soluble in
dim ethylfonnamide, very slightly soluble in methanol,
practically insoluble in ethanol (96 per cent).
It is sensitive to light in dilute solutions.
IDENTIFICATION
First identification: B, D.
Second identification A , C
A. Ultraviolet and visible absorption spectrophotometry
(2.2.25).
L. (2S,5£,6i$-6-[[(2S,5Æ ,6i3-6-[[(2i?)-2-am ino-2-
(4-hydroxyphenyl) acetyl] amino] -3j3-dimethyl-7 -oxo-4-thia-1 - Test solution Dissolve 25 m g in 5 m L of dimethyl sulfoxide R
azabicydo [3.2.0]heptane-2-carbonyl] amino]-3,3-dimethyl-7- and dilute to 50 m L w ith methanol R. Dilute 2 m L o f the
oxo-4-thia-l-azabicydo[3.2.0]heptane-2-carboxylic a d d solution to 200 m L with methanol R.
(6-APA amoxicillin amide). Spectral range 300-450 nm .
PhEur Absorption maxima A t 362 nm , 381 nm and 405 nm .
Absorbance ratios:
— -^ 362^381 = 0.57 to 0.61;
— A^ qi/A^qs — 0.87 to 0.93.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison amphotericin B CRS.
1-168 Amphotericin 2016
If the spectra obtained show differences, dry the substance to adjusted to p H 3.9 using concentrated ammonia R and
be examined and reference substance at 60 °C at a pressure 68 volumes o f acetonitrile R;
not exceeding 0.7 kPa for 1 h and record new spectra.
C. T o 1 m L o f a 0.5 g/L solution in dimethyl sulfoxide R, add Time Mobile phase A Mobile phase B
5 m L o f phosphoric acid R to form a lower layer, avoiding (min) (percent V/V) (per cent V/V)
mixing the 2 liquids. A blue ring is immediately produced at 0 -3 100 0
the junction o f the liquids. Mix, an intense blue colour is
3 -2 3 100-»70 0*30
produced. A dd 15 m L o f water R and mix; the solution
becomes pale yellow. 23 - 33 70-» 0 30-» 100
D. Examine the chromatograms obtained in the test for 3 3 -4 0 0 100
related substances.
Results T he principal peak in the chromatogram obtained
with the test solution at 383 nm is similar in retention time Flow rate 0.8 mL/min.
to the principal peak in the chromatogram obtained with Detection Spectrophotom eter
reference solution (a). — at 303 nm: detection of tetraenes;
— at 383 nm: detection o f heptaenes.
TESTS
Related substances Injection 20 |iL of the test solution and reference
solutions (b), (c), (d) and (e).
Liquid chromatography (2.2.29). Protect the solutions from light
and use within 24 h of preparation, except for reference Identification of impurities Use the chromatograms supplied
solution (c) which should be injected immediately after its with amphotericin B for peak identification CRS and the
preparation. chromatograms obtained with reference solution (e) to
identify the peaks due to impurities A and B.
Solvent mixture 10 g/L solution of ammonium acetate R,
N-metkylpyrrolidone R, methanol R (1:1:2 VIVIV). Relative retention W ith reference to amphotericin B (retention
time = about 16 min): im purity B = about 0.75;
Test solution Dissolve 20.0 m g o f the substance to be
im purity A = about 0.8; nystatin = about 0.85.
examined in 15 m L o f N-methylpyrrolidone R and within 2 h
dilute to 50.0 m L w ith the solvent mixture. Dilute 5.0 m L of System suitability at 383 nm Reference solution (d):
this solution to 25.0 m L with the solvent mixture. — resolution: minimum 1.5 between the 2 peaks presenting a
relative retention o f about 0.7.
Reference solution (a) Dissolve 20.0 mg o f amphotericin B CRS
in 15 m L o f N-methylpyrrolidone R and within 2 h dilute to Limits:
50.0 m L with the solvent mixture. Dilute 5.0 m L of this — impurity A at 303 nm: not more than 2.5 times the area of
solution to 25.0 m L with the solvent mixture. the principal peak in the chromatogram obtained with
reference solution (c) (5.0 per cent); if intended for use in
Reference solution (b) Dilute 1.0 m L o f reference solution (a)
the m anufacture of parenteral preparations: not more
to 100.0 m L with the solvent mixture.
than the area o f the principal peak in the chromatogram
Reference solution (c) Dissolve 20.0 m g o f nystatin CRS in obtained with reference solution (c) (2.0 per cent);
15 m L o f N-methylpyrrolidone R and within 2 h dilute to — any other impurity at 303 nm: for each impurity, not more
50.0 m L with the solvent mixture. D ilute 5.0 m L of the than 0.5 times the area o f the principal peak in the
solution to 25.0 m L with reference solution (a). Dilute chromatogram obtained with reference solution (c)
2.0 m L o f this solution to 100.0 m L with the solvent (1.0 per cent);
mixture. — impurity B at 383 nm: not more than 4 times the area of
Reference solution (d) In order to prepare impurities B and C, the principal peak in the chromatogram obtained with
dissolve 10 m g o f the substance to be examined in 5 m l . of reference solution (b) (4.0 per cent);
N-methylpyrrolidone R and within 2 h add 35 m L o f a mixture — any other impurity at 383 nm: for each impurity, n o t more
of 1 volume o f methanol R and 4 volumes of anhydrous than 2 times the area o f the principal peak in the
ethanol R A dd 0.10 m L o f dilute hydrochloric acid R, mix and chrom atogram obtained with reference solution (b)
incubate at 25 °C for 2.5 h. Add 10 m L of 10 g/L solution (2.0 per cent);
of ammonium acetate R and mhc. — total at 303 and 383 nm: maximum 15.0 per cent;
Reference solution (e) Dissolve 4 m g o f amphotericin B for peak — disregard limit at 303 nm: 0.05 times the area of the
identification CRS (containing impurities A and B) in 5 m l. o f principal peak in the chromatogram obtained with
N-methylpyrrolidone R and within 2 h dilute to 50 m L with reference solution (c) (0.1 per cent);
the solvent mixture. — disregard limit at 383 nm: 0.1 times the area o f the
Blank solution T he solvent mixture. principal peak in the chromatogram obtained with
Column: reference solution (b) (0.1 per cent).
— size. I = 0.15 m , 0 = 4.6 mm; Loss on drying (2.2.32)
— stationary phase: base-deactivated end-capped octadecylsQyl M axim u m 5.0 per cent, d eterm in ed on 1.000 g by drying in
silica gel for chromatography R (3 pm); an oven at 60 °C at a pressure not exceeding 0.7 kPa.
— temperature: 20 °C. Sulfated ash (2.4.14)
Mobile phase: M aximum 3.0 per cent, determ ined on 1.0 g; if intended for
— mobile phase A: mix 1 volume o f methanol R, 3 volumes o f use in the m anufacture o f parenteral preparations: maximum
acetonitrile R and 6 volumes o f a 4.2 g/L solution o f citric 0.5 p er cent.
acid R previously adjusted to p H 4.7 using concentrated Bacterial endotoxins (2.6.14)
ammonia R\ Less than 1.0 IU/mg, if intended for use in the manufacture
— mobile phase B: mix 12 volumes o f methanol R, o f parenteral preparations without a further appropriate
20 volumes of a 4.2 g/L solution o f citric acid R previously procedure for the removal o f bacterial endotoxins.
2016 Ampicillin 1-169
A SSA Y
Protect all solutions from light throughout the assay Dissolve
25.0 m g in dimethyl sulfoxide R and dilute, with shaking* to
25.0 m L with the same solvent. U nder constant stirring of
this stock solution, dilute with dimethyl sulfoxide R to obtain
solutions of appropriate concentrations (the following
concentrations have been found suitable: 44.4, 66.7 and
100 IU /m L). Prepare final solutions by diluting 1:20 with
0.2 M phosphate buffer solution p H 10.5 so that they all
contain 5 per cent V/V o f dimethyl sulfoxide. Prepare the
reference and the test solutions simultaneously. Carry out the
microbiological assay of antibiotics (2.7.2).
STORAGE
C. amphotericin X 2 (13-O-ethyl-amphotericin B).
Protected from light, at a tem perature of 2 °C to 8 °C in an
airtight container. I f the substance is sterile, store in a sterile, PhEur
tam per-proof container.
L A B E L L IN G
T h e label states, where applicable, that the substance is
suitable for use in the manufacture o f parenteral Ampicillin ★ ★
★ ★
preparations.
(Anhydrous Ampicillin, Ph Eur monograph 0167)
IM P U R IT IE S
Specified impurities: A , B.
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). It is therefore n o t necessary to identify these
impurities for dem onstration o f compliance. See also 5.10. CieH iiNsO iS 349.4 69-53-4
Control of impurities in substances for pharmaceutical use): C.
A c tio n a n d u se
Penicillin antibacterial.
P re p a r a tio n s
Ampicillin Capsules
Ampicillin Oral Suspension
PhEur_____________________________
D E F IN IT IO N
(2 S, 5R, 6R)-6- [[(22Q-2-Amino-2-phenylacetyl] amino] -3,3-
dimethyl-7 -oxo-4-thia-1 -azabicyclo [3.2.0] heptane-2-
carboxylic acid.
Semi-synthetic product derived from a fermentation product.
C o n te n t
96.0 per cent to 102.0 p er cent (anhydrous substance).
A. amphotericin A (28,29-dihydro-amphotericin B),
CHARACTERS
A p p e a ra n c e
W hite or almost white, crystalline powder.
S o lu b ility
Sparingly soluble in water, practically insoluble in acetone, in
ethanol (96 per cent) and in fatty oils. It dissolves in dilute
solutions of acids and o f alkali hydroxides.
I t shows polymorphism (5.9).
ID E N T IF IC A T IO N
First identification A , D.
Second identification B, C, D.
A. Infrared absorption spectrophotometry (2.2.24).
Preparation Discs o f potassium bromide R.
B. amphotericin X I (13-O-methyi-amphotericin B), Comparison anhydrous ampicUHn CRS.
B. Thin-layer chromatography (2.2.27).
Test solution Dissolve 25 m g of the substance to be examined
in 10 m L o f sodium hydrogen carbonate solution R.
1-170 Ampicillin 2016
im p u r it ie s
0 > C°2H
V t >< CH3
^ " r r s CHj
H H H . 3-phenylpyrazin-2-ol,
A. (2S,5R, 6i?)-6-amino-3j3-dimethyl-7-oxo-4-thia-1- h nh 2
azabicydo [3.2.0] heptane-2-carboxyHc a d d
° 0 J > C°2H
(6-am inopenidllanic ad d ),
H NH H V n A ^ C H 3
H H
° v _ N*- V\ *^CH3
CH,
II H H
0
G. (3.R,6i?)-3,6-diphenylpiperazine-2j5-dione,
1-172 Ampicillin Sodium 2016
C o n te n t p H (2.2.3)
91.0 per cent to 102.0 per cent (anhydrous substance). 8.0 to 10.0.
Dissolve 2.0 g in carbon dioxide-free water R and dilute to
CHARACTERS
20 m L with the same solvent. M easure 10 min after
A p p e a ra n c e
dissolution.
White or almost white powder, hygroscopic.
Specific o p tic a l ro ta tio n (2.2.7)
Solubility
+ 258 to + 287 (anhydrous substance).
Freely soluble in water, sparingly soluble in acetone,
practically insoluble in fatty oils and in liquid paraffin. Dissolve 62.5 mg in a 4 g/L solution of potassium hydrogen
phthalate R and dilute to 25.0 m L with the same solvent.
ID E N T IF IC A T IO N
R e lated su b s ta n c e s
First ideritification A, D
Liquid chromatography (2.2.29).
Second identification B, C, D
Test sdution (a) Dissolve 31.0 m g o f the substance to be
A. Infrared absorption spectrophotometry (2.2.24). examined in mobile phase A and dilute to 50.0 m L with
Preparation Dissolve 0.250 g in 5 m L of water R, add 0.5 m L mobile phase A.
of dilute acetic add R, swirl and allow to stand for 10 min in Test solution (b) Dissolve 31.0 m g of the substance to be
iced water. Filter the crystals through a small sintered-glass examined in mobile phase A and dilute to 10.0 m L with
filter (40) (2.1.2), applying suction, wash with 2-3 m L o f a mobile phase A. Prepare immediately before use.
mixture of 1 volume o f water R and 9 volumes of acetone R,
then dry in an oven at 60 °C for 30 min.
Reference solution (a) Dissolve 27.0 m g of anhydrous
ampicillin CRS in mobile phase A and dilute to 50.0 m L with
Comparison ampidEin trihydrate CRS. mobile phase A.
B. Thin-layer chromatography (2.2.27). Reference solution (b) Dissolve 2.0 m g of cefradine CRS in
Test solution Dissolve 25 m g o f the substance to be examined mobile phase A and dilute to 50 m L with mobile phase A.
in 10 m L of sodium hydrogen carbonate solution R. T o 5.0 m L o f this solution add 5.0 m L o f reference
Reference solution (a) Dissolve 25 mg of ampidOin solution (a).
trihydrate CRS in 10 m L of sodium hydrogen carbonate Reference solution (c) Dilute 1.0 m L of reference solution (a)
solution R to 20.0 m L with mobile phase A.
Reference solution (b) Dissolve 25 mg of amoxicillin Reference solution (d) T o 0.20 g o f the substance to be
trihydrate CRS and 25 m g of ampidBn tnhydrate CRS in examined add 1.0 m l. of water R. H eat the solution at 60 °C
10 m L of sodium hydrogen carbonate solution R. for 1 h. D ilute 0.5 m L of this solution to 50.0 m L with
Plate TLC sUamsed silica gel plate R. mobile phase A.
Mobile phase M ix 10 volumes of acetone R and 90 volumes of Column:
a 154 g/L solution o f ammonium acetate R previously adjusted — size: I = 0.25 m , 0 = 4.6 mm;
to p H 5.0 with glacial acetic add R. — stationary phase, octadecylsifyl silica gel for chromatography R
Application 1 |iL. (5 pm).
Development Over a path of 15 cm. Mobile phase:
— mobile phase A: mix 0.5 m L o f dilute acetic add R, 50 m L
Drying In air. of 0.2 M potassium dihydrogen phosphate R and 50 m L of
Detection Expose to iodine vapour until the spots appear and acetonitrüe R, then dilute to 1000 m L with water R;
examine in daylight
2016 Ampicillin Sodium 1-173
— mobüe phase B : mix 0.5 m L o f dilute acetic acid R, 50 m L — stationary phase: diatomaceous earth for gas
of 0.2 M potassium ¡Mhydrogen phosphate R and 400 m L o f chromatography R impregnated with 10 per cent mtm of
acetomtrüe R, then dilute to 1000 m L with water R-} macrogol 1000 R.
Carrier gas nitrogen for chromatography R.
Time Mobile phase A Mobile phase B Flow rate 40 mlVmin.
(min) (percent V/V) (per cent V/V) Temperature:
o -f. 85 15 — column: 60 °C;
i, - (f, + 30) 85 -> 0 15-» 100 — irgecdon pore. 100 °C;
— detector. 150 °C.
(fj, + 30) - (fg + 45) 0 100
Detection Flam e ionisation.
(fj, + 45) - (fg + 60) 85 15
Calculate the content o f methylene chloride taking its density
tR= retention time of ampidllin determined with reference solution (c) at 20 °C to be 1.325 g/mL.
Lima:
If the mobile phase composition has been adjusted to achieve — methylene chloride: maximum 0.2 per cent mlm.
the required resolution, the adjusted composition will apply H eav y m e ta ls (2.4.8)
at time zero in the gradient and in the assay. M aximum 20 ppm .
Flow rate 1.0 mL/min. 1.0 g complies with test C . Prepare the reference solution
Detection Spectrophotom eter at 254 nm. using 2 m L o f lead standard solution (10 ppm Pb) R.
Irgecdon 50 (iL o f reference solutions (b) and (c) with W a te r (2.5.12)
isocratic elution at the initial mobile phase composition and M aximum 2.0 per cent, determined on 0.300 g.
50 yL o f test solution (b) and reference solution (d) B a c te ria l en d o to x in s (2.6.14)
according to the elution gradient described under Mobile Less than 0.15 IU/mg, if intended for use in the manufacture
phase; inject mobile phase A as a blank according to the o f parenteral preparations without a further appropriate
elution gradient described under M obile phase. procedure for the removal of bacterial endotoxins.
Identification of peaks Use the chromatogram obtained with A SSAY
reference solution (d) to identify the peaks due to ampicillin
Liquid chromatography (2.2.29) as described in the test for
and am pid llin d im er.
related substances with the following modifications.
Relative retention W ith reference to ampicillin: am pidllin
Mobile phase Initial composition of the mixture o f mobile
dim er = about 2.8.
phases A and B, adjusted where applicable.
System suitability: reference solution (b):
— resolution: m in im u m 3.0 between the peaks due to
Irgecdon T est solution (a) and reference solution (a).
ampicillin and cefradin; if necessary adjust the ratio A:B System suitability: reference solution (a):
of the mobile phase. — repeatability: m axim um relative standard deviation of
1.0 per cent after 6 injections.
Limits:
— ampiaDin dimer, n o t more than 4.5 times the area of the Calculate the percentage content of am pidllin sodium by
principal peak in the chromatogram obtained with multiplying the percentage content of ampicillin by 1.063.
reference solution (c) (4.5 per cent); STO RA G E
— any other impurity: for each impurity, not more than twice In an airtight container. I f the substance is sterile, store in a
the area o f the principal peak in die chromatogram sterile, airtight, tam per-proof container.
obtained with reference solution (c) (2 per cent).
IM P U R IT IE S
iV ,iV -D im ethylaniline (2.4.26, Method B)
M aximum 20 ppm.
O > C°2H
2 -E th y lh ex a n o ic a c id (2.4.28)
V nA xh3
M aximum 0.8 per cent m/m.
M eth y len e c h lo rid e
CHa
H H
Gas chromatography (2.2.28).
Internal standard solution Dissolve 1.0 m L o f ethylene A. (2S,5R, 6/?)-6-amino-3,3-dimethyl-7 -oxo-4-thia-1 -
chloride R in water R and dilute to 500.0 m L with the same azabicydo[3.2.0]heptane-2-carboxylic a d d
solvent. (6-aminopenidllanic a d d ),
Test solution (a) Dissolve 1.0 g o f the substance to be
examined in water R and dilute to 10.0 m L with the same
solvent.
Test solution (b) Dissolve 1.0 g o f the substance to be
examined in water R, add 1.0 m L o f the internal standard
solution and dilute to 10.0 m L with water R.
Reference solution Dissolve 1.0 m L o f methylene chloride R in
water R and dilute to 500.0 m L w ith the same solvent. B. (25,5i?,6i?)-6-[[(25)-2-amino-2-phenylacetyI]amino]-3,3-
T o 1.0 m L o f this solution add 1.0 m L o f the internal dimethyl-7 -oxo-4-thia-1 -azabicyclo [3.2.0] heptane-2-
standard solution and dilute to 10.0 m L with water R. carboxylic a d d (L-ampicillin),
Column:
— material: glass;
— size. / = 1.5 m , 0 = 4 mm;
1-174 Ampicillin Sodium 2016
J. (2S,5i?,6.R)-6-[(2,2-dimethylpropanoyl)amino]-3,3-
dim ethyl-7-oxo-4-thia-l-azabicydo[3.2.0]heptane-2-
C. (45)-2-(3,6-dioxo-5-phenylpiperazin-2-yl)-5,5- carboxylic ad d ,
dimethylthiazolidme-4-caiboxylic a d d (diketopiperazines of
ampidllin), CH3 o
H3C^ _ ^
COjH H3C H NH
H ^ H N - \ .C H 3 CO2H
X ^ Nn î A S CH3
I 0 R
K. (2R)-2-[(2,2-dimethjdpropanoyl)amino]-2-phenylacetic
add,
D . R = C 0 2H : (4<S)-2-[[[(2iQ-2-amino-2-phenylacetyl]
amino]carboxymeth5d]-5,5-dimethylthiazolidine-4-carboxylic
H <NH2
a d d (penicilloic ad d s o f ampidllin),
F. R = H: (2i?5',45)-2-[[[(2Ä)-2-amino-2-phenylacetyl] co2h
amino]methyI]-5,5-dimeth^thiazolidine-4-carboxylic a d d
(penilloic ad d s of ampidllin),
L. (2Æ)-2-amino-2-phenylacetic a d d (D-phenylglycine),
CO2H
E. (2i?)-2-[[[(2S’,5£,6i?)-6-[[(2.R)-2-amino-2-phenylacetyl]
amino]-3,3-dimethyl-7 -oxo-4-thia-1-azabicydo [3.2.0]hept-2-
yl] carbonyl] amino]-2-phenylacetic add (ampicillinyl-D-
phenylglycine),
G . (3Æ,6i?)-3,6-diphenylpiperazme-2,5-dione,
h nh2
I. (2S,5i?,6Æ)-6-[[(2i?)-2-[[(2Æ)-2-amino-2-phenylacetyl]
amino]-2-phenylacetyl] amino]-3,3-dimethjd-7-oxo-4-thia 1-
azabicydo[3.2.0]heptane-2-carboxylic add
(D-phenylglycylampicillin),
2016 Ampicillin Trihydrate 1-175
G . (3i?,6i?)-3,6-diphenylpiperazine-2,5-dione,
A. (2S,5R, 6i?)-6-amino-3,3-dimethyl-7-oxo-4-thia-1-
azabicydo[3.2.0]heptane-2-carboxylic a d d
(6-aminopemdllanic a d d ),
H . 3-phenylpyrazin-2-ol,
H NH,
o J ^ C°2H ★ ★
Amylmetacresol ★ ★
(Ph. Eur. monograph 2405) *****
3 II H H
O
h3c ^ ^oh
J. (2S,5R,6R)-6 -[(2,2-dimethylpropanoyl)amino]-3,3-
ch3
dimethyl-7-oxo-4-thia-1-azabicydo [3.2.0] heptane-2-
carboxylic a d d ,
Ci2H180 178.3 1300-94-3
CH3 o
h3c ^_ Y Action and use
H3C h NH Antiseptic.
CO2H PhEur.
DEFINITION
5-Methyl-2-pentylphenol.
K. (21$-2-[(2,2-dimethylpropanoyl)amino]-2-phenylacetic Content
ad d , 98.0 per cent to 102.0 per cent.
CHARACTERS
Appearance
C lear or alm ost clear liquid, or solid crystalline mass,
colourless or slightly yellow when freshly prepared.
T h e substance changes colour during storage by darkening
and/or discolouration to dark yellow, brownish-yellow or
L. (2jR)-2-amino-2-phenylacetic a d d (D-phenylglycine),
pink.
Solubility
Practically insoluble in water, very soluble in acetone and in
ethanol (96 per cent).
It solidifies at about 22 °C.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Preparation Film between 2 plates of potassium bromide R.
Comparison amylmetacresol CRS.
TESTS
Related substances
Gas chrom atography (2.2.25): use the normalisation
procedure.
M . co-oligomers o f ampicillin and of penidlloic adds of
Internal standard solution Dissolve 0.100 g o f
ampicillin,
butylkydroxytoluene R in 2-propanol R and dilute to 10.0 m l.
with the same solvent.
Test solution (a) Dissolve 0.1000 g of the substance to be
CO2H examined in 2-propanol R and dilute to 10.0 m L with the
sam e solvent.
Test solution (b) T o 2.0 m L of test solution (a) add 2.0 m L of
the internal standard solution and dilute to 10.0 m L with
2-propanol R.
N . (3iS)-6-[[(2Æ)-2-amino-2-phenylacetyl] amino]-2,2-
dimethyl-7-oxo-2,3,4,7-tetrahydro-l,4-thiazepine-3-carboxylic
Reference solution (a) Dissolve 10 m g of m-cresol R
(impurity B) and 10 mg o f p-cresol R (impurity D ) in
ad d .
2-propanol R and dilute to 100.0 m L with the same solvent.
PhEur
Reference solution (b) Dissolve the contents o f a vial of
amylmetacresol for peak identification CRS (containing
impurities A, G an d K) in 1.0 m l o f 2-propanol R.
Reference solution (c) Dissolve 0.1000 g of amylmetacresol CRS
in 2-propanol R and dilute to 10.0 m L with the same solvent.
T o 2.0 m L o f this solution add 2.0 m L o f the internal
standard solution and dilute to 10.0 m L with 2-propanol R.
Reference solution (d) Dilute 1.0 m L of test solution (a) to
100.0 m L w ith 2-propanol R. Dilute 1.0 m L of this solution
to 20.0 m L with 2-propanol R
Column:
— material: fused silica;
— size. I = 30 m , 0 = 0.25 mm;
1-178 Amylmetacresol 2016
Time Temperature
(min) rc)
Column 0-17.5 100-»240 B. 3-methylphenol (m-cresol),
17.5 - 32.5 240
H3C' ^ \ ^ 0H
Injection port 250 || H CH3 and enantiomer
Detector 250
C. 5-methyl-2-[(2i?S)-2-methyIbutyl]phenol,
Detection Flam e ionisation.
Injection 1.0 jiL o f test solution (a) and reference
solutions (a), (b) and (d).
Identification of impurities Use the chromatogram supplied
with amylmetacresolfor peak identification CRS and the
chrom atogram obtained with reference solution (b) to
identify the peaks due to impurities A, G and K.
Relative retention W ith reference to amylmetacresol (retention
time = about 16 min): impurity G
(diastereoisomer 1) = about 0.51; impurity G
(diastereoisomer 2) = about 0.53; impurity D = about 0.77;
O
im purity B = about 0.78; impurity K = about 0.95;
impurity A = about 0.99.
E. 1-(2-hydroxy-4-methylphenyl)pentan-1-one,
System suitability: reference solution (a):
— resolution: m inim um 1.5 between the peaks due to
impurities D and B.
Limits:
— impurity A: maximum 0.6 per cent;
— impurities G (sum o f the 2 diastereoisomers), K. for each O
impurity, maximum 0.15 p er cent;
— unspecified impurities: for each impurity, maximum F. 1-(2-hydroxy-5-methylphenyl)pentan-1-one,
0.10 per cent;
— total: maximum 1.0 per cent;
— disregard limit, the area of the peak due to amylmetacresol
in the chrom atogram obtained with reference solution (d)
(0.05 per cent).
S u lfa te d a s h (2.4.14)
M aximum 0.1 per cent, determined on 1.0 g.
A SSA Y
Gas chrom atography (2.2.28) as described in the test for o
related substances with the following modification.
Injection 1.0 jiL o f test solution (b) and reference solution (c). H . ethyl pentanoate,
Calculate the percentage content o f C 12H i80 from the
declared content of amylmetacresol CRS.
STORAGE
In an airtight, non-metallic container, protected from lig h t
IM P U R IT IE S I. 3-methylphenyl pentanoate,
Specified impurities A, G , K
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the monograph. T hey are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general m onograph Substances for pharmaceutical use
J. 4-methylphenyl pentanoate,
(2034). It is therefore no t necessary to identify these
impurities for dem onstration o f compliance. See also 5.10. K. unknow n structure.
Control of impurities in substances for pharmaceutical use): B, C, _____________________________________________________ _ PhEur
D, E, F, H, I, J.
2016 Anastrozole 1-179
Mobile phase'.
Anastrozole ***** — mobile phase A: phosphoric add R, water for
*+ +* chromatography R (0.1:100 V/V)',
(Ph Eur monograph 2406) *
— mobile phase B: phosphoric add R, acetonitrUe R l
CN (0.1:100 V/V)',
Column-. H3C - L
— size. I = 0.15 m , 0 = 4.6 mm; h T CN
— stationary phase, end-capped, polar-embedded octadecylsUyl
amorphous organosUica polymer R (3.5 nm). A. 2-[3-[(litS )-l-cyanoethyl]-5-(l/f-l,2,4-triazol-l-
ylmethyl) phenyl] -2-methylpropanenitrile,
1-180 Antazoline Hydrochloride 2016
I. 2 ,2 '-[5-(chloromethyl)benzene-1,3-diyI]
H3C -7 ^
bis(2-methylpropanenitrile).
PhEur
C. 2 , 2 [ 5 -(bromomethyl)benzene-1,3-diyi]
bis (2-methylpropanenitrile),
HCI
D . 2 ,2 '-[5-(dibromomethyl)benzene-1,3-diyi]
bis (2-methylpropanenitrile),
A c tio n a n d u se
Histamine H I receptor antagonist] antihistamine.
PhEur__________________________________________________________
D E F IN IT IO N
Antazoline hydrochloride contains n o t less than 99.0 per cent
E. 2 , 2 [5-(hydroxymethyl)benzene-1,3-diyI] and not more than the equivalent o f 101.0 per cent o f
bis(2-methylpropanenitrile), N-benzyl-N-[(4,5-dihydro-lH-irnidazol-2-yl)methyl]aniline
hydrochloride, calculated w ith reference to the dried
substance.
CHARACTERS
A white or almost white, crystalline powder, sparingly soluble
in water, soluble in alcohol, slightly soluble in methylene
F. 4-methylbenzenesulfonic ad d , chloride.
I t melts at about 240 °C, with decomposition.
ID E N T IF IC A T IO N
First identification A, D.
Second identification B, C, D.
A Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
antazoline hydrochloride CRS. Examine the substances as discs
G. 2 ,2 '-[5-(4H-1,2,4-triazol-4-ylmethyl) benzene-1,3-diyI] prepared using potassium chloride R.
bis (2-methylpropanenitrile), B. Examine the chromatograms obtained in the test for
related substances in daylight after spraying. T he principal
spot in the chromatogram obtained with test solution (b) is
similar in position, colour and size to the principal spot in
the chromatogram obtained with reference solution (b).
C. T o 5 m L o f solution S (see Tests) add, drop by drop,
dilute sodium hydroxide solution R until an alkaline reaction is
2016 Apomorphine Hydrochloride Hemihydrate 1-181
produced. Filter. T h e precipitate, washed with two 1 m L of 0.1 M alcoholic potassium hydroxide is equivalent to
quantities, each o f 10 m L, o f water R and dried in a 30.18 m g o f C 17H 20CIN 3.
desiccator under reduced pressure, melts (2.2.14) at 119 °C IM P U R IT IE S
to 123 °C.
D . It gives reaction (a) of chlorides (2.3.1).
TESTS
S o lu tio n S
Dissolve 2.0 g in carbon dioxide-free water R prepared from
distilled water R, heating at 60 °C if necessary. Allow to cool
and dilute to 100 m L with the same solvent.
A p p e a ra n c e o f so lu tio n A. N - (2-aminoethyI) -2-(benzylphenylamino) acetamide.
Solution S is clear (2.2.1) and not more intensely coloured
................... PhEur
than reference solution (2.2.2, Method II).
A cidity o r a lk a lin ity
T o 10 m L o f solution S add 0.2 m L o f methyl red solution R.
N ot more than 0.1 mT. o f 0.01 M hydrochloric add or 0.01 M
sodium hydroxide is required to change the colour of the Apomorphine Hydrochloride *****
indicator.
Hemihydrate *****
R elated su b s ta n c e s
(Ph. Eur. monograph 0136)
Examine by thin-layer chromatography (2.2.27), using silica
gel GF2 5 4 R as the coating substance. H eat the plate at
110 °C for 15 m in before using.
Test solution (a) Dissolve 0.10 g of the substance to be
examined in methanol R and dilute to 5 m L with the same
solvent.
Test solution (b) D ilute 1 m L o f test solution (a) to 5 m L
with methanol R.
C nH ^C IN C V /iH zO 312.8 41372-20-7
Reference solution (a) Dilute 0.5 m L o f test solution (a) to
100 m L with methanol R. A c tio n a n d u se
Reference solution (b) Dissolve 20 mg o f antazoUne D opam ine receptor agonist; treatm ent o f Parkinson’s disease.
hydrochloride CRS in methanol R and dilute to 5 m L with the
P r e p a r a tio n
same solvent.
Apomorphine Hydrochloride for Homoeopathic Preparations
Reference solution (c) Dissolve 20 mg o f xylometazoUne
hydrochloride CRS in 1 m L o f test solution (a) and dilute to PhFtr __________________________________________
5 m L with methanol R. D E F IN IT IO N
Apply to the plate 5 jiL o f each solution. Develop over a (6aR)-6-M ethyl-5,6,6a,7-tetrahydro-4ii-dibenzo[d«^]
path of 15 cm using a mixture of 5 volumes of quinoline- 10,11 -diol hydrochloride h em ih ydrate.
dietkylamme R, 10 volumes of methanol R and 85 volumes of
C o n te n t
ethyl acetate R. D ry the plate in a current o f warm air for
98.5 p er cent to 101.5 p e r cent (dried substance).
15 min. Examine in ultraviolet light at 254 nm . T he test is
not valid unless the chromatogram obtained with reference CHARACTERS
solution (c) shows two clearly separated prindpal spots. A p p e a ra n c e
Spray with a mixture of equal volumes of a 200 g/L solution W hite or slightly yellowish-brown or green-tinged greyish,
of ferric chloride R and a 5 g/L solution of potassium crystalline pow der or crystals; on exposure to air and light,
ferruyanide R. Examine immediately in daylight. Any spot in the green tinge becomes more pronounced.
the chrom atogram obtained with test solution (a), apart from S o lu b ility
the prindpal spot, is not more intense than the spot in the Sparingly soluble in water and in ethanol (96 per cent),
chromatogram obtained with reference solution (a) practically insoluble in toluene.
(0.5 per cent).
ID E N T IF IC A T IO N
H eavy m e ta ls (2.4.8)
First identification B, D
1.0 g complies with test C for heavy metals (20 ppm).
Prepare the reference solution using 2 m L o f lead standard Second identification A, C, D
solution (10 ppm Pb) R. A. Ultraviolet and visible absorption spectrophotometry
(2.2.25).
Loss o n d ry in g (2.2.32)
N ot more than 0.5 per cent, determined on 1.000 g by Test solution Dissolve 10.0 mg in a 10.3 g/L solution of
drying in an oven at 105 °C for 3 h. hydrochloric add R and dilute to 100.0 m L with the same a d d
solution. Dilute 10.0 m L of the solution to 100.0 m L with a
S ulfa te d a s h (2.4.14)
10.3 g/L solution o f hydrochloric add R.
N ot more than 0.1 per cent, determined on the residue
obtained in the test for loss on drying. Spectral range 230-350 nm
Absorption maximum A t 273 nm.
ASSAY
Dissolve 0.250 g in 100 m L of alcohol R. A dd 0.1 m L of
Shoulder A t 300-310 nm.
phenolphthalein solution R l. Titrate with 0.1 M alcoholic Spedfic absorbance at the absorption maximum 530 to 570.
potassium hydroxide. B. Infrared absorption spectrophotom etry (2.2.24).
1-182 Apomorphine Hydrochloride Hemihydrate 2016
2 - 32 85 -> 68 15 -* 32
32-37 68 32
2016 Aprotinin 1-183
CHARACTERS
Appearance
Almost white hygroscopic powder.
Solubility
Soluble in water and in isotonic solutions, practically
insoluble in organic solvents.
B. 7,8-didehydro-4,5a-epoxy-17-methyimorphinan-3,6a-diol IDENTIFICATION
(m orphine). A. Thin-layer chromatography (2.2.27).
Test solution Solution S (see Tests).
Reference solution D ilute aprotinin solution BRP in water R to
obtain a concentration of 15 Ph. Eur. UVmL.
Plate TLC silica gel G plate R.
Mobile phase water R, glacial acetic acid R (80:100 V/V)
co n taining 100 g/L o f sodium acetate R.
Application 10 |iL.
Development Over a path o f 12 cm.
Drying In air.
C. (6aR)-9-[7,8-didehydro-4,5a-epoxy-3-hydroxy-17-
Detection Spray with a solution of 0.1 g of ninhydrin R in a
m ethylm orphinan- 6a-yl] -6-methyl-5,6 , 6 a, 7-tetrahydr o-4H-
mixture o f 6 m L o f a 10 g/L solution of cupric chloride R,
dibenzo [de,g\ quinoline- 10, 11-diol (m orphine-apomorphine
21 m L of glacial acetic acid R and 70 m L o f anhydrous
dimer).
ethanol R. Dry the plate at 60 °C.
__________________________________________________________ PhEi*
Results T he principal spot in the chromatogram obtained with
the test solution is similar in position, colour and size to the
principal spot in the chromatogram obtained with the
reference solution.
Aprotinin ***** B. Determine the ability of the substance to be examined to
*+ ** inhibit trypsin activity using the m ethod described below.
(Ph Eur monograph 0580) *
Test solution Dilute 1 m L o f solution S to 50 m L w ith buffer
H - A rg-Pro - Asp - P h e -C y s -L e u -G lu -P r o -P r o -T y r -
solution pH 7.2 R.
-------------------------------------- . t(J
Trypsin solution Dissolve 10 mg of trypsin BRP in 0.002 M
T h r-G ly -P ro -C y s -L y s -A )a -A rg — lie— lie—Arg-
I______________________20 hydrochloric add and dilute to 100 m L with the same acid.
Tyr - Phe - Tyr - Asn - Ala - Lys - Ala - Gly - Leu - Cys -
_____________________________ m Casein solution Dissolve 0.2 g of casein R in buffer solution
G ln -T h r-P h e -V a l-T y r -G ly -G ly -C y s -A rg -A la — pH 7.2 R and dilute to 100 m L w ith the same buffer
I_____________ i 2_____
Lys - Arg - Asn - Asn - Phe - Lys - Ser - Ala - Glu - Asp -
solution.
_I 50 Predpitating solution gladd acetic add R, water R, anhydrous
Cys - Met - Arg - Thr - Cys - Gly - Gly - Ala - OH
----------------------- - 58 ethanol R (1:49:50 V/V/V).
Mix 1 m L o f the test solution with 1 m L o f the trypsin
C 284H 432N 84O79S7 6511 solution. Allow to stand for 10 m in and add 1 m L o f the
casein solution. Incubate at 35 °C for 30 m in. Cool in iced
A ctio n a n d u se
water and add 0.5 m L o f the precipitating solution. Shake
Antifibrinolytic. and allow to stand at room temperature for 15 min.
PhEtr__________________________________________________________ T h e solution is cloudy. Carry out a blank test under the
same conditions using buffer solution pH 7.2 R instead of the
D E F IN IT IO N test solution. T he solution is not cloudy.
Aprotinin is a polypeptide consisting of a chain of 58 amino
acids. It inhibits stoichiometrically the activity of several
TESTS
proteolytic enzymes such as chymotrypsin, kallikrein, plasmin Solution S
and trypsin. It contains not less than 3.0 Ph. Eur. U . Prepare a solution o f the substance to be examined
of aprotinin activity per milligram, calculated with reference containing 15 Ph. Eur. UVmL, calculated from the activity
to the dried substance. stated on the label.
P R O D U C T IO N
Appearance of solution
Solution S is clear (2.2.1).
T he animals from which aprotinin is derived must fulfil the
requirem ents for the health of animals suitable for hum an Absorbance (2.2.25)
consumption. Maximum 0.80 by measuring at the absorption maximum at
277 nm.
T he m ethod o f manufacture is validated to demonstrate that
the product, if tested, would comply with the following tests. Prepare a solution o f the substance to be examined
containing 3.0 Ph. Eur. U ./ mL.
A b n o rm a l to x ic ity (2.6.9)
Inject into each mouse a quantity o f the substance to be Des-Ala-aprotinin and des-Ala-des-Gly-aprotinin
examined containing 2 Ph. Eur. U. dissolved in a sufficient Capillary zone electrophoresis (2.2.47): Use the normalisation
quantity o f waterfor injections R to give a volume o f 0.5 mL. procedure.
H is ta m in e (2.6.10)
M axim um 0.2 |ig o f histamine base p er 3 Phi Eur. U .
1-184 Aprotinin 2016
Test solution Prepare a solution o f the substance to be dihydrate R and 66.07 g o f ammonium sulfate R in
ex am in ed inwater R containing n o t less than 1000 m L of water; filter and degas;
1 Ph. E ur. U ./m L.
Reference solution D ilute aprotinin solution BRP in water R to Time Mobile phase A Mobile phase B
obtain the same concentration as the test solution. (min) (percent V/V) (percent V/V)
Capillary: 0 -2 1 92 64 8*36
— material: uncoated fused silica; 21 -3 0 64 0 3 6 * 100
— sizer, effective length = 45-60 cm, 0 = 75 Jim.
Temperature 25 °C.
Flow rate 1.0 mlVmin.
CZE buffer Dissolve 8.21 g o f potassium ¿¡hydrogen
Detection Spectrophotom eter a t 210 nm .
phosphate R in 400 m L o f water R, adjust to p H 3.0 with
phosphoric add R, dilute to 500.0 m L with water R and filter Irgection 40 |xL.
through a m em brane filter (nominal pore size 0.45 Jim). Relative retention W ith reference to aprotinin (retention
Detection Spectrophotom eter a t 214 nm . tim e = 17.0 min to 20.0 min): impurity C = about 0.9.
Between-run rinsing Rinse the capillary for at least 1 min with System suitability: reference solution:
— resolution: minimum 1.5 betw een the peaks due to
0.1 M sodium hydroxide filtered through a m em brane filter
impurity C and aprotinin;
(nominal pore size 0.45 Jim) and for 2 min with the C ZE
—■symmetry factor, maximum 1.3 for the peak due to
buffer.
aprotinin.
Irgection U nder pressure or vacuum (for example, 3 s a t a
differential pressure o f 3.5 kPa).
Limits:
— impurity C: m aximum 1.0 p er cent;
Migration Apply a field strength o f 0.2 kV/cm, using the C ZE — any other impurity: m axim um 0.5 p er cent;
buffer as the electrolyte in b oth buffer reservoirs. — sum of impurities other than C: m aximum 1.0 p er c e n t
Run time 30 min.
Aprotinin oligomers
Identification of impurities Use the electropherogram supplied Size-exclusion chromatography (2.2.30): U se the
with aprotinin solution BRP and the electropherogram norm alisation procedure.
obtained with the reference solution to identify the peaks due
Test solution Prepare a solution o f the substance to be
to impurities A and B.
examined in water R containing about 5 Ph. Eur. UVmL.
Relative migration W ith reference to aprotinin (migration Reference solution T reat the substance to be examined to
tim e = about 22 m in): impurity A = about 0.98;
obtain about 2 per cent aprotinin oligomers. F or example,
im purity B = about 0.99.
heat freeze-dried aprotinin at about 110 °C for about 4 h.
System suitability Reference solution after at least 6 injections: T h en dissolve in water R to obtain a concentration o f about
— migration time: aprotinin = 19.0 m in to 25.0 m in ; 5 Ph. Eur. UVmL.
— resolution: m inim um 0.8 between the peaks due to
Column 3 columns coupled in series:
impurities A and B; minimum 0.5 between the peaks due
— size. I = 0.30 m , 0 = 7.8 m m ;
to im purity B and aprotinin;
— stationary phase, hydrophilic silica gelfor chromatography R
— peak distribution: the electrophoregram obtained is
o f a grade suitable for fractionation o f globular proteins in
qualitatively and quantitatively sim ilar to the
the relative molecular mass range o f 20 000 to
electropherogram supplied with aprotinin solution BRP,
10 000 000 (8 jam).
— height of the prindpal peak: at least 1000 times the height
o f the baseline noise. If necessary, adjust the sample load Mobile phase acetomtrUe R, glacial acetic add R, water R
to give peaks o f sufficient height. (2:2:6 V/VIV)-, filter and degas.
Limits: Flow rate 1.0 mlVmin.
— impurity A: maximum 8.0 per cent; Detection Spectrophotom eter at 277 nm .
— impurity B: m axim um 7.5 per cent. Irgection 100 |iL.
Pyroghitamyl-aprotinin and related compounds Run lime 40 min.
L iquid chrom atography (2.2.29): use the normalisation Relative retention W ith reference to aprotinin m onom er
procedure. (retention time = 24.5 min to 25.5 min): aprotinin
Test solution Prepare a solution of the substance to be dim er = about 0.9.
e x a m in ed in mobile phase A, containing about System suitability: reference solution:
5 Ph. E ur. UVmL. — resolution: minimum 1.3 betw een the peaks due to
Reference sdudon Dissolve the contents of a vial o f aprotinin aprotinin dim er and monom er;
for system suitability CRS in 2.0 m L o f mobile phase A. — symmetry factor, maximum 2.5 for the peak due to
Column: aprotinin m onom er.
— size. I = 0.075 m , 0 = 7.5 mm; Limit:
— stationary phase, strong cation-exchange silica gelfor — total: maxim um 1.0 p er cent.
chromatography R (10 pm); Loss on drying (2.2.32)
— temperature. 40 °C. Maximum 6.0 per cent, determ ined on 0.100 g by drying
Mobile phase: in vacuo.
— mobile phase A: dissolve 3.52 g o f potassium dihydrogen
B a c te r ia l e n d o to x in s (2.6.14)
phosphate R and 7.26 g o f disodium hydrogen phosphate Less than 0.14 IU per European Pharm acopoeia U nit o f
dihydrate R in 1000 m L o f water; filter and degas; aprotinin, if intended for use in the m anufacture of
— mobile phase B: dissolve 3.52 g o f potassium dihydrogen
parenteral preparations w ithout a further appropriate
phosphate R, 7.26 g o f disodium hydrogen phosphate procedure for the removal o f bacterial endotoxins.
2016 Aprotinin 1-185
ASSAY L A B E L L IN G
T he activity of aprotinin is determined by measuring its The label states:
inhibitory action on a solution of trypsin o f known activity. — the num ber o f European Pharmacopoeia Units of
T he inhibiting activity of the aprotinin is calculated from the aprotinin activity per milligram;
difference between the initial activity and the residual activity — where applicable, that the substance is suitable for use in
of die trypsin. the m anufacture o f parenteral preparations.
T he inhibiting activity of aprotinin is expressed in European IM P U R IT IE S
Pharmacopoeia Units. 1 Ph. Eur. U . inhibits 50 p er cent of
the enzymatic activity of 2 micro katals of trypsin. Ra - Arg - Pro - A s p -P h e -C y s -L e u -G lu -P ro -P ro - Tyr -
___ !_________________ | to
Use a reaction vessel with a capacity of about 30 m L, T h r-G ly -P ro -C y s -L y s -A la -A rg — lie— lie—Arg -
provided with: I «
Tyr - Phe - Tyr - Asn - Ala - Lys - Ala - Gly - Leu - Cys -
— a device that will maintain a temperature of 25 ± 0.1 ° Q _____________________________ m
— a stirring device, such as a magnetic stirrer; Gin - Thr - Ptie - Val - Tyr - Gly - Gly - Cys - Arg - Ala-
I w
— a lid with 5 holes for accommodating the electrodes, the Lys - Arg - Asn - Asn - Phe - Lys - Ser - Ala - Glu - Asp -
tip of a burette, a tube for the admission of nitrogen and —i 30
Cys - M e t-A rg -T h r-C y s -G ly -R b
the introduction of the reagents. ----------------------------- «
An automatic o r manual titration apparatus may be used.
In the latter case the burette is graduated in 0.05 m L and the A. Ra = H , Rb = OH: aprotinin-(1-56)-peptide,
pH -m eter is provided with a wide reading scale and glass and
B. Ra = H , Rb = Gly-OH: aprotinin-(l-57)-peptide,
calomel or glass-silver-silver chloride electrodes.
C. Ra = Glp, Rb = Gly-Ala-OH: (5-oxoprolyl)aprotinin
Test solution Prepare a solution of the substance to be (pyroglutamylaprotinin).
examined in 0.0015 M borate buffer solution pH 8.0 R
expected to contain 1.67 Ph. Eur. UVmL (about 0.6 mg __________________________________________________ PhEur
(m mg) per millilitre).
Trypsin solution Prepare a solution o f trypsin BRP containing
about 0.8 micro katals per millilitre (about 1 mg/mL), using
0.001 M hydrochloric acid as the solvent. Use a freshly Aprotinin Concentrated Solution
prepared solution and keep in iced water.
Trypsin and aprotinin solution T o 4.0 m L o f the trypsin (Ph Eur monograph 0579) *
solution add 1.0 m L of the test solution. Dilute immediately
H - Arg - Pro - Asp - Phe - Cys - Leu - Glu - Pro - Pro - Tyr-
to 40.0 m L with 0.0015 M borate buffer solution pH 8.0 R
Allow to stand a t room tem perature for 10 min and then T h r-G ly -P ro -C y s -L y s -A la -A rg — lie— lie—Arg-
keep in iced water. Use within 6 h o f preparation. |_________________ 20
T y r -P h e -T y r-A s n -A la -L y s -A la -G ly -L e u -C y s -
Dilute trypsin solution Dilute 0.5 m L of the trypsin solution to ________________________________________ 221
Gin - Thr - Phe - Val - Tyr - Gly - Gly - Cys - Arg - Ala -
10.0 m L with 0.0015 M borate buffer solution pH 8.0 R Allow
to stand at room tem perature for 10 min and then keep in Lys - Arg - Asn - Asn - Phe - Lys - Ser - Ala - Glu - Asp -
—i 30
iced water. Cys - Met - Arg- Thr - Cys - Gly - Gly - Ala - OH
M aintain an atm osphere o f nitrogen in the reaction flask and
stir continuously; introduce 9.0 m L o f 0.0015 M borate buffer
solution pH 8.0 R and 1.0 m L of a freshly prepared 6.9 g/L C 284H 432N 84O79S7 6511
solution of benzqylarginine ethyl ester hydrochloride R. Adjust to
p H 8.0 with 0.1 M sodium hydroxide. W hen the tem perature A c tio n a n d u se
has reached equilibrium at 25 ± 0.1 °C, add 1.0 m L of die Antifibrinolytic.
trypsin and aprotinin solution and start a timer. M aintain at PhEur_________________________________________________________
p H 8.0 by the addition of 0.1 M sodium hydroxide and note
the volume added every 30 s. Continue the reaction for D E F IN IT IO N
6 m in . D eterm ine the num ber of m illilitres of 0.1 M sodium Aprotinin concentrated solution is a solution o f aprotinin, a
hydroxide used p er second (nx mL). Carry out, under the polypeptide consisting of a chain of 58 amino adds, which
same conditions, a titration using 1 .0 m L of the dilute inhibits stoichiometrically the activity o f several proteolytic
trypsin solution. Determ ine the num ber o f millilitres of enzymes such as chymotrypsin, kallikrem, plasmin and
0.1 M sodium hydroxide used per second (n2 mL). trypsin. It contains not less than 15.0 Ph. Eur. U.
Calculate the aprotinin activity in European Pharmacopoeia o f aprotinin activity per millilitre.
Units per milligram using the following expression: P R O D U C T IO N
T h e animals from which aprotinin is derived m ust fulfil the
4000 (2na — ni) requirem ents for the health of anim als suitable for hu m an
m consumption.
T h e m ethod of manufacture is validated to demonstrate that
T he estimated activity is n o t less than 90 per cent and not the product, if tested, would comply with the following tests.
more than 110 per cent o f the activity stated on the label. A b n o rm a l to x icity (2.6.9)
Inject into each mouse a quantity of the preparation to be
ST O R A G E
examined containing 2 Ph. Eur. U . diluted with a suffident
In an airtight, tam per-proof container, protected from light.
quantity of water for injections R to give a volume of 0.5 mL.
H ista m in e (2.6.10)
M aximum 0.2 |ig o f histamine base per 3 Ph. Eur. U.
1-186 Aprotinin 2016
Reference solution D ilute aprotinin solution BRP in water R to Detection Spectrophotom eter at 214 nm .
obtain a concentration o f 15 Ph. Eur. U ./m L. Between-run rinsing Rinse the capillary for at least 1 min with
Plate TLC silica gel G plate R. 0.1 M sodium hydroxide filtered through a m em brane filter
(n o m in al pore size 0.45 Jim) and for 2 m in with the C ZE
Mobile phase water R, glacial acetic acid R (80:100 V/V)
buffer.
containing 100 g/L o f sodium acetate R.
Injection U nder pressure or vacuum (ibr example, 3 s at a
Application 10 jiL.
differential pressure o f 3.5 kPa).
Development Over a path of 12 cm.
Migration Apply a field strength o f 0.2 kV/cm, using the C ZE
Drying In air. buffer as the electrolyte in b oth buffer reservoirs.
Detection Spray with a solution of 0.1 g o f mnhydrin R in a Run time 30 min.
m ixture o f 6 m L o f a 10 g/L solution o f cupric chloride R,
Identification of impurities Use the electropherogram supplied
21 m L o f glacial acetic acid R and 70 m L o f anhydrous
with aprotinin solution BRP and die electropherogram
ethanol R. D ry the plate at 60 °C.
obtained with the reference solution to identify the peaks due
Results T he principal spot in the chromatogram obtained with to impurities A and B.
the test solution is similar in position, colour and size to the
Relative migration W ith reference to aprotinin (migration
principal spot in the chrom atogram obtained with the
tim e = about 22 m in): impurity A = about 0.98;
reference solution.
im purity B = about 0.99.
B. D eterm ine the ability o f the preparation to be examined to System suitability Reference solution after at least 6 injections:
inhibit trypsin activity using the m ethod described below. — migration time, aprotinin = 19.0 m in to 25.0 min;
Test solution D ilute 1 m L o f solution S to 50 m L with buffer — resolution: m in im u m 0.8 between the peaks due to
solution pH 7.2 R impurities A and B; minim um 0.5 between the peaks due
Trypsin solution Dissolve 10 m g of trypsin BRP in 0.002 M to im purity B and aprotinin;
hydrochloric add and dilute to 100 m L with the same acid. — peak distribution: the electrophoregram obtained is
Casein solution Dissolve 0.2 g o f casein R in buffer solution qualitatively and quantitatively similar to the
pH 7.2 R and dilute to 100 m L with the same buffer electropherogram supplied with aprotinin solution BRP,
solution. — height of the principal peak: at least 1000 times the height
o f the baseline noise. I f necessary, adjust the sample load
Predpitating solution glacial acetic add R, water R, anhydrous
to give peaks o f a sufficient height.
ethanol R (1:49:50 VfVIV).
Limits.
M ix 1 m L o f the test solution with 1 m L o f the trypsin
— impurity A: m axim um 8.0 per cent;
solution. Allow to stand for 10 min and add 1 m L of the
— impurity B: maximum 7.5 p er cent.
casein solution. Incubate at 35 °C for 30 min. Cool in iced
w ater and add 0.5 m L of the precipitating solution. Shake Pyroglutamyl-aprotinin and related compounds
and allow to stand at room tem perature for 15 min. Liquid chrom atography (2.2.29): use die normalisation
T h e solution is cloudy. Carry out a blank test under the procedure.
sam e conditions using buffer solution pH 7.2 R instead o f the Test solution D ilute th e preparation to be examined in mobile
test solution. T h e solution is n o t cloudy. phase A to a concentration o f about 5 Ph. Eur. UVmL.
TESTS Reference solution Dissolve the contents o f a vial o f aprotinin
S o lu tio n S for system suitability CRS in 2.0 m L o f mobile phase A.
Prepare a solution c o n tain in g 15 Ph. Eur. U./m L, if Column:
necessary by dilution, on the basis o f the activity stated on — size. I = 0.075 m , 0 = 7.5 mm ;
the label. — stationary phase, strong cation-exchange silica gel for
A p p e a ra n c e o f so lu tio n
chromatography R (10 pm);
Solution S is clear (2.2.1). — temperature. 40 °C.
Mobile phase.
A b so rb a n c e (2.2.25)
— mobile phase A: dissolve 3.52 g o f potassium dihydrogen
M axim um 0.80 by measuring at the absorption m m m n m at
phosphate R and 7.26 g o f disodium hydrogen phosphate
277 nm .
dihydrate R in 1000 m L o f water; filter and degas;
Prepare a solution containing 3.0 Ph. Eur. IJ /mT.. — mobile phase B: dissolve 3.52 g o í potassium dihydrogen
D e s -A la -a p ro tm in a n d d e s-A la -d e s-G ly -a p ro tin m phosphate R, 7.26 g o f disodium hydrogen phosphate
Capillary zone electrophoresis (2.2.47) U se the normalisation dihydrate R and 66.07 g of ammonium sulfate R in
procedure. 1000 m L o f water; filter and degas;
Test solution D ilute the preparation to be examined in water R
to obtain a concentration o f n o t less than 1 P h Eur. U./m L. Time Mobile phase A Mobile phase B
Reference solution D ilute aprotinin solution BRP in water R to (min) (percent V/V) (percent V/V)
obtain the same concentration as the test solution. 0 -2 1 92 -»64 8*36
Capillary: 2 1 -3 0 64*0 3 6 * 100
— material: u nfoated fused silica;
2016 Aprotinin 1-187
Flow rate 1.0 mL/min. T he inhibiting activity of the aprotinin is calculated from the
Detection Spectrophotom eter at 210 nm. difference between the initial activity and the residual activity
o f the trypsin.
Irgection 40 (iL.
T h e inhibiting activity of aprotinin is expressed in European
Relative retention W ith reference to aprotinin (retention
Pharmacopoeia Units. 1 Ph. Eur. U . inhibits 50 per cent o f
time = 17.0 m in to 20.0 m in); impurity C = about 0.9.
the enzymatic activity o f 2 microkatals of trypsin.
System suitability: reference solution:
— resolution: minim um 1.5 between the peaks due to U se a reaction vessel with a capacity o f about 30 mL,
impurity C and aprotinin; provided with:
— symmetry factor, maximum 1.3 for the peak due to — a device th at will maintain a tem perature of 25 + 0.1 °C;
— a stirring device, such as a magnetic stirrer;
aprotinin.
— a lid with 5 holes for accommodating the electrodes, the
Limits:
tip o f a burette, a tube for the admission of nitrogen and
— impurity C: maximum 1.0 per cent;
the introduction o f the reagents.
— any other impurity, maximum 0.5 per cent; -
— sum of impurities other than C: maximum 1.0 per cent. A n autom atic o r manual titration apparatus may be used.
In the latter case the burette is graduated in 0.05 m L and the
Aprotinin oligomers pH -m eter is provided with a wide reading scale and glass and
Size-exclusion chromatography (2.2.30) Use the calomel or glass-silver-silver chloride electrodes.
normalisation procedure.
Test solution W ith 0.0015 M borate buffer solution pH 8.0 R
Test solution Dilute the preparation to be examined in water R prepare an appropriate dilution (D) of the aprotinin
to obtain a concentration of about 5 Ph. E ur. UVmL.
concentrated solution expected, on the basis of the stated
Reference solution T reat the substance to be examined to potency, to contain 1.67 Ph. Eur. IJ VmT -
obtain about 2 p er cent aprotinin oligomers. F or example, Trypsin solution Prepare a solution o f trypsin BRP containing
heat freeze-dried aprotinin at about 110 °C for about 4 h.
about 0.8 microkatals per millilitre (about 1 mg/mL), using
T hen dissolve in water R to obtain a concentration o f about
0.001 M hydrochloric acid as the solvent. Use a freshly
5 Ph. E ur. U./mL. prepared solution and keep in iced water.
Column 3 columns coupled in series: Trypsin and aprotinin solution T o 4.0 m L of the trypsin
— sizer. I = 0.30 m , 0 = 7.8 mm; solution add 1.0 m L o f the test solution. Dilute immediately
— stationary phase: hydrophilic silica gel for chromatography R
to 40.0 m L with 0.0015 M borate buffer solution pH 8.0 R.
o f a grade suitable for fractionation of globular proteins in
Allow to stand at room tem perature for 10 min and then
the relative molecular mass range of 20 000 to keep in iced water. U se within 6 h of preparation.
10 000 000 (8 nm).
Dilute trypsin solution Dilute 0.5 m L of the trypsin solution to
Mobile phase acetordtrUe R, glacial acetic add R, water R 10.0 m L with 0.0015 M borate buffer solution pH 8.0 R. Allow
(2:2:6 VIV/V); filter and degas. to stand at room temperature for 10 min and then keep in
Flow rate 1.0 mL/min. iced water.
Detection Spectrophotom eter at 277 nm. Maintain an atmosphere of nitrogen in the reaction flask and
Irgection 100 (iL. stir continuously; introduce 9.0 m L of 0.0015 M borate buffer
Run time 40 min. solution pH 8.0 R and 1.0 m L of a freshly prepared 6.9 g/L
solution of benzoylarginine ethyl ester hydrochloride R.'A djust to
Relative retention W ith reference to aprotinin m onom er
p H 8.0 with 0.1 M sodium hydroxide. W hen the tem perature
(retention time = 24.5 min to 25.5 min): aprotinin
has reached equilibrium at 25 ± 0.1 °C, add 1.0 m L of the
dimer = about 0.9.
trypsin and aprotinin solution and start a timer. M aintain at
System suitability, reference solution: p H 8.0 by the addition of 0.1 M sodium hydroxide and note
— resolution: m in im u m 1.3 between the peaks due to
the volume added every 30 s. Continue the reaction for
aprotinin dim er and monomer;
6 min. Determine the num ber o f millilitres of 0.1 M sodium
— symmetry factor, m axim u m 2.5 for the peak due to
hydroxide used per second ( ^ m L). Carry out, under the
aprotinin m onom er. same conditions, a titration using 1.0 mL of the dilute
Limit: trypsin solution. Determine the num ber of millilitres of
— total: maxim um 1.0 per cen t 0.1 M sodium hydroxide used per second (n2 mL).
Specific activity o f the dry residue Calculate the aprotinin activity in European Pharmacopoeia
Minimum 3.0 Ph. Eur. U. o f aprotinin activity per milligram U nits per millilitre vising the following expression:
of dry residue.
Evaporate 25.0 m l, to dryness in a water-bath, dry the 4000 (2ri2 — n i )
residue at 110 °C for 15 h and weigh- From the mass of the m
residue and the activity determined as described below,
calculate the nu m b er o f European Pharmacopoeia U nits per
D = dilution factor of the aprotinin concentrated
milligram of dry residue.
solution to be examined in order to obtain a
Bacterial endotoxins (2.6.14) solution containing 1.67 Ph. Eur. UVmL.
Less than 0.14 IU per European Pharmacopoeia U nit of
aprotinin, if intended for use in the manufacture of T h e estimated activity is not less than 90 per cent and not
parenteral preparations without a further appropriate m ore than 110 per cent of the activity stated on the label.
procedure for the removal o f bacterial endotoxins. STO RA G E
ASSAY In an airtight, tam per-proof container, protected from light.
T he activity of aprotinin is determined by measuring its
inhibitory action on a solution of trypsin of.known activity.
1-188 Arachis Oil 2016
TESTS
Arginine Hydrochloride }***%
S o lu tio n S
(Ph Eur. monograph 0805) * Dissolve 2.5 g in distilled water R and dilute to 50 m L with
the same solvent.
H H* -NH2 A p p e a ra n c e o f so lu tio n
• HCI Solution S is clear (2.2.1) and not m ore intensely coloured
than reference solution BY6 (2.2.2, Method II).
NH
S p ecific o p tic a l ro ta tio n (2.2.7)
+ 21.0 to + 23.5 (dried substance).
C eH uC m A 210.7 1119-34-2
Dissolve 2.00 g in hydrochloric add R1 and dilute to 25.0 m L
A c tio n a n d u se with the same ad d .
Amino add; nutrient. N in h y d rin -p o sitiv e su b s ta n c e s
P r e p a r a tio n Amino a d d analysis (2.2.56). F or analysis, use M ethod 1.
Arginine Hydrochloride Infusion T h e concentrations o f the test solution and die reference
Arginine Hydrochloride Oral Suspension solutions may be adapted according to the sensitivity o f the
Sterile Arginine Hydrochloride C oncentrate equipm ent used. T h e concentrations o f all solutions are
adjusted so that the system suitability requirem ents described
PhEur___________________________________________________________ in general chapter 2.2.46 are fulfilled, keeping the ratios o f
D E F IN IT IO N concentrations betw een all solutions as described.
(25)-2-Amino-5-guanidinopentanoic a d d hydrochloride. Solution A water R ot a sample preparation buffer suitable for
Ferm entation product, extract or hydrolysate o f protein. the apparatus used.
Test solution Dissolve 30.0 m g o f the substance to be
C o n te n t
examined in solution A and dilute to 50.0 m L with
98.5 per cent to 101.0 per cent (dried substance).
solution A.
CHARACTERS Reference solution (a) D ilute 1.0 m L o f the test solution to
A p p e a ra n c e 100.0 m L with solution A. D ilute 2.0 m L of this solution to
W hite or alm ost white, crystalline pow der o r colourless 10.0 m L with solution A.
crystals.
Reference solution (b) Dissolve 30.0 m g o f proUne R in
S o lu b ility solution A and dilute to 100.0 m L with solution A. Dilute
Freely soluble in w ater, very slightly soluble in ethanol 1.0 m L o f the solution to 250.0 m L with solution A.
(96 per cent). Reference solution (c) Dilute 6.0 m L o f ammonium standard
ID E N T IF IC A T IO N solution (100 ppm N H J R to 50.0 m L with solution A. D ilute
First identification A , B, E 1.0 m L o f this solution to 100.0 m L with solution A.
Second identification A , C, D} E Reference solution (d) Dissolve 30 m g of isoleucine R and
A. Specific optical rotation (see Tests). 30 m g o f leucine R in solution A and dilute to 50.0 m L with
solution A. Dilute 1.0 m L o f the solution to 200.0 m L with
B. Infrared absorption spectrophotom etry (2.2.24).
solution A.
Comparison arginine hydrochloride CRS.
Blank solution Solution A.
C . Thin-layer chromatography (2.2.27).
Inject suitable, equal amounts o f the test, blank and reference
Test solution Dissolve 10 m g o f the substance to be examined solutions into die amino add analyser. Rim a program
in water R and dilute to 50 m L with the same solvent. suitable for the determ ination o f physiological amino adds.
Reference solution. Dissolve 10 m g o f arginine System suitability Reference solution (d):
hydrochloride CRS in water R and dilute to 50 m L with the — résolution: minimum 1.5 between die peaks due to
sam e solvent. isoleucine and leucine.
Plate TLC silica gel plate R. Calculation of percentage contents:
Mobile phase concentrated ammonia R, 2-propanol R — for any ninhydrin-positive substance detected at 570 nm ,
(30:70 VIV). use die concentration o f arginine in reference solution (a);
Application 5 fiL. — for any ninhydrin-positive substance detected at 440 nm ,
use d ie concentration o f proline in reference solution (b);
Development Over 2/3 of the plate.
if a peak is above the reporting threshold at both
Drying At 105 °C until the am m onia disappears completely. wavelengths, use the result obtained at 570 n m for
Detection Spray with mnhydrin solution R and heat at 105 °C quantification.
for 15 min. Limits:
Results T he p rindpal spot in the chrom atogram obtained with — any ninhydrin-positive substance: for each impurity,
the T est solution is similar in position, colour and size to the m axim um 0.2 p er cent;
p rindpal spot in the chrom atogram obtained with the — total: maximum 0.5 per cent;
reference solution. — reporting threshold: 0.05 p er cent.
D . Dissolve about 25 m g in 2 m L o f water R. Add 1 m L of T h e thresholds indicated under Related substances
a-naphthol solution R and 2 m L o f a mixture o f equal volumes (Table 2034.-1) in the general m onograph Substances for
o f strong sodium hypochlorite solution R and water R. A red pharmaceutical use (2034) do n o t apply.
colour develops.
E. I t gives reaction (a) o f chlorides (2.3.1).
2016 Argon 1-193
S u lfates (.2.4.13)
M axim um 300 ppm.
Dilute 10 m L o f solution S to 15 m L w ith distilled water R.
A m m onium
B. (2S)-2-amino-5-(carbamoylamino)pentanoic a d d
A m ino acid analysis (2.2.56) as described in the test for
(dtrulline),
ninhydrin-positive substances with the following
modifications.
Irgection T est solution, reference solution (c) and blank H,N,
solution.
Limit.
C. (2S)-2,5-diaminopentanoic a d d (ornithine).
— ammonium at 570 nm: not more than the area of the
corresponding peak in the chromatogram obtained with PhEur
reference solution (c) (0.02 per cent), taking into account
the peak due to am m onium in the chromatogram
obtained w ith the blank solution.
I r o n (2.4.9) Argon *****
★ ★
M aximum 10 ppm.
(Ph Eur monograph 2407) *****
In a separating funnel, dissolve 1.0 g in 10 m L of dilute
hydrochloric acid R. Shake with 3 quantities, each of 10 mL, Ar 39.95 7440-37-1
of methyl tsobutyl ketone R l, shaking for 3 min each time. PhEur_____________________________
T o the combined organic layers add 10 m L o f water R and
shake for 3 min. U se the aqueous layer. D E F IN IT IO N
Gas obtained by fractional distillation of ambient air.
H eav y m e ta ls (2.4.8)
M axim u m 10 ppm. C o n te n t
M inim um 99.995 per cent V/V o f Ar, calculated by
Dissolve 2.0 g in water R and dilute to 20 m L with the same
solvent. 12 m L o f the solution complies with test A. Prepare deduction of the sum of impurities found when performing
the reference solution using lead standard solution the test for impurities and the water content.
(1 ppm Pb) R. T his monograph applies to argon for medicinal use.
L oss o n d ry in g (2.2.32) CHARACTERS
M axim um 0.5 per cent, determined on 1.000 g by drying in A p p e a ra n c e
an oven at 105 °C. Colourless gas.
S u lfa te d a s h (2.4.14) S o lu b ility
M axim um 0.1 per cent, determined on 1.0 g. A t 20 °C and at a pressure of 101 kPa, 1 volume dissolves in
about 29 volumes of water.
A SSA Y
Dissolve 0.180 g in 3 m L of anhydrous formic acid R. ID E N T IF IC A T IO N
A dd 30 m L o f anhydrous acetic acid R. T itrate with 0.1 M A. Verify that the gas is not oxygen using a paramagnetic
perchloric acid, determining the end-point potentiometrically analyser (2.5.27).
(2. 2. 20). B. Gas chromatography (2.2.28).
1 m L of 0.1 M perchloric acid is equivalent to 21.07 mg of Gas to be examined T he substance to be examined.
C e H is C lN ^ .
Reference gas Use the following mixture of gases in argon R l:
STORA GE methane R l (5 ppm V/V), nitrogen R l (5 ppm V/V), oxygen R
Protected from light. (5 ppm V/V).
IM P U R IT IE S Column:
— material: stainless steel;
Other detectable impurities (the following substances would, if
— sizer. I = 2 m, 0 = 3 mm;
present at a sufficient level, be detected by one or other of
— stationary phase: molecular sieve for chromatography R
the tests in the monograph. They are limited by the general
(particle size 150-180 nm, pore size 0.5 nm).
acceptance criterion for other/unspecified impurities. It is
therefore not necessary to identify these impurities for Carrier gas helium for chromatography R.
dem onstration o f compliance. See also 5.10. Control of Flow rate 10 mL/min.
impurities in substances for pharmaceutical use): A , B, C. Temperature:
— column: 50 °C;
— detector. 150 °C.
H,N Detection Thermal conductivity.
Irgection 25 JiL.
A. (2S)-2,6-diaminohexanoic a d d (lysine), System suitability. Reference gas:
— resolution: minim um 3.0 between the peaks due to
argon/oxygen and nitrogen and minimum 2.0 between
the peaks due to nitrogen and methane.
Results T h e prindpal peak in the chromatogram obtained
with the gas to be examined is similar in retention time to
the prindpal peak in the chromatogram obtained with the
reference gas.
1-194 Aripiprazole 2016
TESTS if* it
if ★
Im p u ritie s
Aripiprazole ★ ★
Gas chromatography (2.2.28). (Ph. Eur. monograph 2617) *****
Gas to be examined T h e substance to be exam in ed.
Reference gas U se the following mixture o f gases in argon Rl:
methane R l (5 ppm VIV), nitrogen R l (5 ppm VIV), oxygen R
(5 ppm VIV).
Column•
— material: stainless steel;
— size. I = 4 m , 0 = 4 m m ;
— stationary phase, molecular sieve for chromatography R
(particle size 150-180 pm , pore size 0.5 nm ). C23H27CI2N3O2 448.4 129722-12-9
Carrier gas argon R l.
Flow rate 70 mL/min. A c tio n a n d u se
D opam ine D 2 receptor antagonist; neuroleptic
Temperature.
— column: 80 °C; PhEur___________________________________________________________
— detector. 40 °C.
D E F IN IT IO N
Detection Discharge ionisation.
7-[4- [4- (2,3-Dichlorophenyl)piperazin-1-yl] butoxy]-3,4-
Injection 1 m l- dihydroquinolin-2(lii)-one.
Sample rate 100 mL/min. C o n te n t
Relative retention W ith reference to im purity C (retention 98.0 per cent to 102.0 per cent (dried substance).
tim e = about 4.7 min): im purity A = about 0.4;
CHARACTERS
impurity B = about 0.7.
A p p e a ra n c e
System suitability: Reference gas:
W hite or alm ost white crystals o r crystalline powder.
— resolution: m in im u m 3.0 between the peaks due to
impurities A and B and m in im u m 2.0 between the peaks S o lu b ility
due to impurities B and C. Practically insoluble in water, soluble in methylene chloride,
Limits: very slighty soluble in ethanol (96 per cent).
— impurity A: not m ore than the area of the corresponding It shows polym orphism (5.9).
peak in the chrom atogram obtained with the reference gas ID E N T IF IC A T IO N
(5.0 p pm V!V)\ Infrared absorption spectrophotom etry (2.2.24).
— total: m axim um 0.0040 p er cent o f the sum of the areas
Comparison aripiprazole CRS.
o f all the peaks (40.0 p pm VIV).
If the spectra obtained in the solid state show differences,
W a te r (2.5.28)
dissolve the substance to be examined and the reference
M axim um 10.0 ppm VIV, d eterm ined u sin g an electrolytic
substance separately in methylene chloride R, evaporate to
hygrometer.
dryness and record new spectra using the residues.
STORAGE
TESTS
In gaseous or liquid state, in suitable con tainers, complying
A p p e a ra n c e o f so lu tio n
w ith the legal regulations.
I f intended for use in the m anufacture o f parenteral
IM P U R IT IE S preparations, the solution is clear (2.2.1) and n o t more
Specified impurities A, D intensely coloured than reference solution GY 5 (2.2.2,
Other detectable impurities B, C. Method II).
A. oxygen, Dissolve 0.5 g in a mixture o f 10 volumes o f acetic acid R and
90 volumes o f anhydrous ethanol R and dilute to 20 m L with
B. nitrogen,
the same mixture of solvents. Sonicate for about 15 m in,
C. m ethane, shaking occasionally, until dissolution is complete.
D . water.
R e la te d su b s ta n c e s
------ ----------------------------------------------------------------------------- PhEur L iquid chrom atography (2.2.29). Protect the solutions from
light.
Solvent mixture acetic add R, methanol R, acetonitrile R,
water R (1:10:30:60 VIVIVIV).
Test solution Dissolve 50.0 m g o f the substance to be
exam in ed in the solvent mixture and dilute to 50.0 m L with
the solvent mixture. D ilute 5.0 m L o f the solution to
50.0 m L with the solvent mixture.
Reference solution (a) Dilute 1.0 m L o f the test solution to
100.0 m L with the solvent mixture. D ilute 1.0 m L o f this
solution to 10.0 m L with the solvent mixture.
Reference solution (b) Dissolve 5 m g o f the substance to be
exam in e d and 5 m g of aripiprazole impurity F CRS in the
solvent m ixture and dilute to 100 m L with the solvent
2016 Aripiprazole 1-195
2 - 10 80 ->65 20-»35
2 0 -2 5 10 90
A. 7-hydroxy-3,4-dihydroquinolirt-2(lfi)-one,
HOzC L1 H CH
CH3
Relative retention W ith reference to articaine (retention
tim e = about 9 min): impurity A = about 0.8; and enantiomer
Second identification A , C, D
nh 2 A. Ultraviolet and visible absorption spectrophotom etry
(2.2.25).
ch3 Test solution Dissolve 0.10 g in water R and dilute
immediately to 100.0 m L with the same solvent. Add 1.0 m L
o f this solution to 10 m L o f 0.1 M hydrochloric add and
I. methyl 3-am ino-4-methylthiophene-2-caiboxylate
dilute to 100.0 m L with water R
(3-aminoarticaine),
Absorption maximum At 243 nm , determ ined immediately
after dissolution.
i Spedfic absorbance at the absorption maximum 545 to 585.
Ch3 and enantiomer B. Infrared absorption spectrophotom etry (2.2.24).
Comparison ascorbic add CRS.
CH3
C. p H (2.2.3): 2.1 to 2.6 for solution S (see Tests).
— stationary phase: aminopropylsifyl silica gel for Adjust the zero of the apparatus using 0.1 M nitric add.
chromatography R (5 nm)i H eav y m e ta ls (2.4.8)
— temperature: 45 °C. M aximum 10 ppm.
Mobile phase Phosphate buffer solutionj acetonitrUe R1 Dissolve 2.0 g in water R and dilute to 20 m L with the same
(25:75 VIV). solvent. 12 m L of the solution complies with test A. Prepare
Flow rate 1.0 mlVmin. the reference solution using lead standard solution
Detection Spectrophotom eter at 210 nm. (1 ppm Pb) R.
Injection 20 |iL o f the test solution and reference solutions (b) S u lfa te d a s h (2.4.14)
and (c). M aximum 0.1 per cent, determined on 1.0 g.
Run time 2.5 times the retention time of ascorbic acid. A SSAY
Identification of impttrities Use the chromatogram obtained Dissolve 0.150 g in a mixture of 10 m L of dilute sulfuric
with reference solution (b) to identify the peaks due to add R and 80 m L of carbon dioxide-free water R. Add 1 m L of
impurities C and D. starch solution R. Titrate with 0.05 M iodine until a persistent
Relative retention W ith reference to ascorbic acid (retention violet-blue colour is obtained.
time = about 11 min): impurity D = about 0.4; 1 m L of 0.05 M iodine is equivalent to 8.81 mg of CgHgOg.
impurity C = about 1.7.
STORA GE
System suitability. In a non-metallic container, protected from light.
— resolution: minim um 3.0 between the peaks due to
ascorbic a d d and impurity C in the chromatogram IM P U R IT IE S
obtained with reference solution (c); Specified impurities C, D , E
— signal-to-noise ratio: m in im u m 20 for the peak due to Other detectable impurities (the following substances would, if
impurity C in the chromatogram obtained with reference present at a suffident level, be detected by one or other o f
solution (b). the tests in the monograph. They are limited by the general
Limits: acceptance criterion for other/unspecified impurities and/or
— impurities C, D: for each impurity, n o t more than by the general monograph Substances for pharmaceutical vise
1.5 times the area of the corresponding peak in the (2034). It is therefore not necessary to identify these
chrom atogram obtained with reference solution (b) impurities for demonstration of compliance. See also 5.10.
(0.15 per cent); Control of impurities in substances for pharmaceutical use): A , F,
— unspecified impurities: for each impurity, n o t more than the G, H.
area o f the peak due to ascorbic a d d in the
chromatogram obtained with reference solution (b) /° V -C H O
(0.10 per cent); i 1
— total of impurities other than C and D: not more than twice
the area of the peak due to ascorbic a d d in the
A. 2-furaldehyde,
chromatogram obtained with reference solution (b)
(0.2 per cent);
OH oh o
— disregard limic. 0.5 times the area of the peak due to
ascorbic a d d in the chromatogram obtained with
reference solution (b) (0.05 per cent).
rV/™
OH OH O
C opper
M aximum 5 ppm . C. D-xy/o-hex-2-ulosonic a d d (D-sorbosonic ad d ),
Atomic absorption spectrometry (2.2.25, Method I).
OH OH o
Test solution Dissolve 2.0 g in 0.1 M nitric add and dilute to
25.0 m L with the same ad d .
Reference solutions Prepare the reference solutions (0.2 ppm , OH OH O
0.4 ppm and 0.6 ppm ) by diluting copper standard solution
(10 ppm Cu) R w ith 0.1 M nitric add. D . methyl D-xyfo-hex-2-ulosonate (methyl D-sorbosonate),
Source C opper hollow-cathode lamp.
Wavelength 324.8 nm.
Atomisation device Air-acetylene flame.
Adjust the zero o f the apparatus using 0.1 M nitric add.
V" o
Ir o n
M aximum 2 ppm . E. oxalic ad d ,
Atomic absorption spectrometry {2.2.23, Method I).
Test solution Dissolve 5.0 g in 0.1 M nitric add and dilute to
25.0 m L with th e same ad d .
Reference solutions Prepare the reference solutions (0.2 ppm,
0.4 ppm and 0.6 ppm ) by diluting iron standard solution HO OH
(20 ppm Fe) R w ith 0.1 M nitric acid.
Source Iron hollow-cathode lamp. F. (5R)-5-[(lR)-l,2-dihydroxyethyl]-3,4-dihydroxyfuran-
Wavelength 248.3 nm. 2 (5/i)-one,
Atomisation device Air-acetylene flame.
1-200 Ascorbyl Palmitate 2016
A c tio n a n d u se
Excipient.
X x™ 1 , H20
h2n ' ^ ^ v co2h
PhEur.
C4H8N20 3iH 20 150.1 5794-13-8
D E F IN IT IO N
(25)-2-[(2i?)-3,4-Dihydroxy-5-oxo-2,5-dihydrofuraii-2-yl]-2- A c tio n a n d u se
hydroxyethyl hexadecanoate. Amino add.
C o n te n t
PhEur_____________
98.0 per cent to 100.5 per cent (dried substance).
D E F IN IT IO N
CHARACTERS
(2S)-2,4-Diam ino-4-oxobutanoic a d d m onohydrate.
A p p e a ra n c e
W hite or yellowish-white powder. C o n te n t
99.0 per cent to 101.0 p er cent (dried substance).
S o lu b ility
Practically insoluble in water, freely soluble in ethanol CHARACTERS
(96 p er cent) and in m ethanol, practically insoluble in A p p e a ra n c e
methylene chloride and in fatty oils. W hite or almost white, crystalline pow der or colourless
crystals.
ID E N T IF IC A T IO N
A. Specific optical rotation (see Tests). S o lu b ility
Slightly soluble in water, practically insoluble in ethanol
B. Infrared absorption spectrophotom etry (2.2.24).
(96 per cent) and in methylene chloride.
Comparison ascorbyl palmitate CRS.
ID E N T IF IC A T IO N
C. Dissolve about 10 mg in 5 m L o f methanol R.
First identification A , B
T he solution decolourises dichlorophenolindophenol standard
solution R. Second identification A , C
A. Specific optical rotation (see Tests).
TESTS
S o lu tio n S B. Infrared absorption spectrophotom etry (2.2.24).
Dissolve 2.50 g in methanol R and dilute to 25.0 m L with the Comparison asparagine monohydrate CRS.
same solvent C. Examine the chrom atograms obtained in the test for
A p p e a ra n c e o f so lu tio n ninhydrin-positive substances.
Solution S is clear (2.2.1) a n d n o t m ore intensely coloured Results T h e principal spot in th e chrom atogram obtained with
than reference solution BY4 (2.2.2, Method I). test solution (b) is similar in position, colour and size to the
2016 Aspartame 1-201
principal spot in the chromatogram obtained with reference organic phases w ith 10 m L of water R for 3 min.
solution (c). T h e aqueous phase complies with th e limit test for iron.
TESTS H eav y m e ta ls (2.4.8)
S o lu tio n S M aximum 10 ppm .
Dissolve with heating 2.0 g in carbon dioxide-free water R and Dissolve 2.0 g in a mixture of 3 m L of dilute hydrochloric
dilute to 100 m L with the same solvent. acid R and 15 m L of water R with gentle warming if
A p p e a ra n c e o f so lu tio n necessary. Dilute to 20 m L with water R. 12 m L o f the
Solution S is d ear (2.2.1) and colourless (2.2.2, Method II). solution complies with test A. Prepare the reference solution
using lead standard solution (1 ppm Pb) R.
p H (2.2.3)
4.0 to 6.0 for solution S. L oss o n d ry in g (2.2.32)
10.5 per cent to 12.5 p er cent, determined on 1.000 g by
S p ecific o p tic a l ro ta tio n (2.2.7)
drying in an oven at 130 °C for 3 h.
+ 33.7 to + 36.0 (dried substance).
S u lfa te d a sh (2. 4.14)
Dissolve 2.50 g in a 309.0 g/L solution o f hydrochloric acid R
M aximum 0.1 per cent, determined on 1.0 g.
and dilute to 25.0 m L with the same acid.
N in h y d rin -p o sitiv e su b sta n ce s A SSA Y
Thin-layer chromatography (2.2.27). Dissolve 0.110 g in 5 m L of anhydrous formic acid R.
Add 50 m L of anhydrous acetic acid R. T itrate with 0.1 M
Test solution (a) Dissolve 0.25 g of the substance to be
perchloric acid, determining the end-point potentiometrically
examined in water R, heating to not more than 40 °Cj and
( 2 .2.20).
dilute to 10 m L w ith the same solvent.
1 m L of 0.1 M perchloric acid is equivalent to 13.21 mg
Test solution (b) Dilute 1 m L of test solution (a) to 10 m L
o fC 4 H 8N 20 3.
w ith water R.
Reference solution (a) Dilute 1.0 m L o f test solution (a) to IM P U R IT IE S
200 m L with water R. Specified impurities: A, B.
Reference solution (b) Dissolve 25 mg o f glutamic acid R in
H nh2
water R, add 1 m L o f test solution (a) and dilute to 10 m L
w ith water R.
Reference solution (c) Dissolve 25 mg of asparagine
monohydrate CRS in water R and dilute to 10 m L with the A. (25)-2-aminobutanedioic a d d (aspartic ad d ),
sam e solvent.
Hate TLC silica gel G plate R. h nh 2
V
Mobile phase glacial acetic acid R, water R, butanol R ho2c / S s ^ n‘co2h
(25:25:50 V/VIV).
Application 5 pL. B. (2S)-2-aminopentanedioic a d d (glutamic ad d ).
Development Over h alf of the plate. PhEur
Drying A t 110 °C for 15 min.
Detection Spray with ninhydrin solution R and heat at 110 °C
for 10 min.
System suitability: reference solution (b): ★★* * ★
— the chromatogram shows 2 clearly separated principal
Aspartame ★ ★
spots. (Ph Eur monograph 0973) *****
Limit: test solution (a):
— any impurity, any spot, apart from the principal spot, is
not more intense than the principal spot in the
chromatogram obtained with reference solution (a)
(0.5 per cent).
C h lo rid e s (2.4.4)
M axim um 200 ppm .
D ilute 12.5 m L o f solution S to 15 m L with water R.
S u lfa te s (2.4.II)
M axim um 200 ppm. Q 4H 18N 2O 5 294.3 22839-47-0
L oss o n d ry in g (2.2.32) ★ ★
Maxim vim 4.5 per cent, determined on 1.000 g by drying in
Aspartic Acid ★ ★
★ ★
an oven at 105 °C. (Ph. Eur. monograph 0797) *★ *
S u lfa te d a sh (2.4.14)
M aximum 0.2 per cent, determined on 1.0 g. H NH 2
ASSAY
Dissolve 0.250 g in 1.5 m L o f anhydrous formic acid R and
60 m L o f anhydrous acetic add R T itrate immediately with C4H7N 0 4 133.1 56-84-8
0.1 M perchloric add, determining the end-point
potentiometrically (2.2.20). A c tio n a n d u se
1 m L of 0.1 M perchloric acid is equivalent to 29.43 mg Amino a d d .
Of C 14H 18N 2O 5.
P hB r_____________
STORAGE
D E F IN IT IO N
In an airtight container.
Aspartic a d d contains not less th an 98.5 per cent and not
IM P U R IT IE S m ore than the equivalent of 101.5 p e r cent of
Specified impurities A, C (25)-2-am inobutanedioic add, calculated with reference to
Other detectable impurities (the following substances would, if the dried substance.
present at a sufficient level, be detected by one or other of CHARACTERS
the tests in the m onograph. They are limited by the general A white o r almost white, crystalline powder or colourless
acceptance criterion for other/unspecified impurities and/or crystals, slightly soluble in water, practically insoluble in
by the general m onograph Substances for pharmaceutical use alcohol. I t dissolves in dilute mineral adds and in dilute
(2034). I t is therefore not necessary to identify these solutions o f alkali hydroxides.
impurities for dem onstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): B. ID E N T IF IC A T IO N
First identification A, C.
Second identification A , B, D.
A. Specific optical rotation (see T ests).
COzH B. A suspension of 1 g in 10 m L o f water R is strongly a d d
(2.2.4).
C. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with aspartic
A. 2-[(25',53)-5-benzyl-3,6-dioxopiperazm-2-yl]acetic ad d . add CRS. Examine the substances prepared as discs.
D . Examine the chromatograms obtained in the test for
ninhydrin-posTtive substances. T h e prindpal spot in the
chrom atogram obtained with test solution (b) is similar in
position, colour and size to the principal spot in the
chrom atogram obtained with reference solution (a).
CO2H
TESTS
HOjC A p p e a ra n c e o f so lu tio n
Dissolve 0.5 g in 1 M hydrochloric acid and dilute to 10 m L
w ith the same ad d . T he solution is clear (2.2.1) and not
B. (35)-3-amino-4- [[(13)-1 -carboxy-2-phenylethyI] amino] -4-
more intensely coloured than reference solution BY 6 (2.2.2,
oxobutanoic a d d (a-L-aspartyl-L-phenylalanine),
Method II).
S p ecific o p tic a l ro ta tio n (2.2.7)
Dissolve 2.000 g in hydrochloric acid R l and dilute to
25.0 m L with the same add. T h e specific optical rotation is
+ 24.0 to + 26.0, calculated with reference to the dried
substance.
C . (2S)-2-amino-3-phenylpropanoic a d d (L-phenylalanine).
N in h y d rin -p o sitiv e su b stan ces
PhEir
Examine by thin-layer chromatography (2.2.27), using a
TLC silica gel plate R.
Test solution (a) Dissolve 0.10 g o f th e substance to be
exam in e d in 2 m L of ammonia R and dilute to 10 m L w ith
water R.
Test solution (b) Dilute 1 mL of test solution (a) to 50 m L
with water R.
Reference solution (a) Dissolve 10 m g of aspartic add CRS in
2 m L of dilute ammonia R l and dilute to 50 m L with
water R.
Reference solution (b) Dilute 5 m L o f test solution (b) to
20 m L w ith water R.
1-204 Aspirin 2016
a
Effervescent Soluble Aspirin Tablets
Gastro-resistant Aspirin Tablets
Aspirin and Caffeine Tablets
limit test for sulfates (300 ppm ). Carry out the evaluation o f Co-codaprin Tablets
the test after 30 min. Dispersible Co-codaprin Tablets
A m m o n iu m PhEur.
2.4.1) 50 mg complies with limit test B (200 ppm ). Prepare
the standard using 0.1 m L o f ammonium standard solution D E F IN IT IO N
(100 ppm N H J R. 2-(Acetyloxy)benzoic acid.
Ir o n (2.4.9) C o n te n t
In a separating funnel, dissolve 1.0 g in 10 m L o f dilute 99.5 per cent to 101.0 per cent (dried substance).
hydrochloric add R. Shake with 3 quantities, each of 10 mT, CHARACTERS
of methyl isobutyl ketone R l, shaking for 3 min each time. A p p e a ra n c e
T o d ie combined organic layers add 10 m L o f water R and W hite or almost white, crystalline pow der or colourless
shake for 3 min. T he aqueous layer complies with the limit crystals.
test for iron (10 ppm ).
S o lu b ility
H eav y m e ta ls (2.4.8) Slighdy soluble in water, freely soluble in ethanol
2.0 g complies with test D (10 ppm ). Prepare the reference (96 per cent).
solution using 2 m L o f lead standard solution (10 ppm Pb) R.
mp
L oss o n d ry in g (2.2.32) About 143 °C (instantaneous method).
N ot m ore than 0.5 per cent, determined on 1.000 g by
drying in an oven at 105 °C. ID E N T IF IC A T IO N
First identification A, B
Sulfa ted ash (2.4.14)
N ot m ore than 0.1 per cent, d eterm ined on 1.0 g. Second identification B, C, D
A Infrared absorption spectrophotometry (2.2.24).
A SSA Y
Dissolve 0.100 g in 50 m L o f carbon dioxide-free water R, with
Comparison acetylsalicylic add CRS.
slight heating if necessary. Cool and add 0.1 m L o f B. T o 0.2 g add 4 m L o f dilute sodium hydroxide solution R
bromothymol blue solution R l. T itrate with 0.1 M sodium and boil for 3 min. Cool and add 5 mT. o f dilute sulfuric
hydroxide until the colour changes from yellow to blue. add R. A crystalline precipitate is formed. Filter, wash the
precipitate and dry at 100-105 °C. T h e melting point
1 m L o f 0.1 M sodium hydroxide is equivalent to 13.31 m g of
C4H7N 0 4. (2.2.14) is 156 °C to 161 °C.
C. In a test tube mix 0.1 g with 0.5 g o f caldum hydroxide R.
STO RA G E
H eat the mixture and expose to the fumes produced a piece
Protected from light. o f filter paper impregnated with 0.05 m L o f nitrobenzaldehyde
PhEur solution R. A greenish-blue o r greenish-yellow colour develops
on the paper. M oisten the paper with dilute hydrochloric
add R T h e colour becomes blue.
D . Dissolve with heating about 20 m g o f the precipitate
obtained in identification test B in 10 m L o f water R and
cool. T h e solution gives reaction (a) o f salicylates (2.3.1).
TESTS
A p p e a ra n c e o f so lu tio n
T h e solution is clear (2.2.1) and colourless (2.2.2,
Method 11).
2016 Aspirin 1-205
Dissolve 1.0 g in 9 m L o f ethanol (96 per cent) R. Prepare the reference solution using lead standard solution
Related substances (1 ppm Pb) obtained by diluting lead standard solution
L iquid chromatography (2.2.29). Prepare the solutions (100 ppm Pb) R with a mixture o f 6 volumes of water R and
immediately before use. 9 volumes of acetone R.
Injection 10 |iL.
h o 2c CO2H
Run time 7 times the retention time o f acetylsalicylic ad d .
Identification of impurities U se the chrom atogram obtained OH
w ith reference solution (a) to identify the peak due to
im purity C; use the chromatogram supplied with B. 4-hydroxybenzene-1,3-dicarboxylic a d d
acetylsalicylic acid for peak identification CRS and the (4-hydroxyisophthalic ad d ),
chrom atogram obtained with reference solution (c) to identify
the peaks due to impurities A, B, D , E and F.
COjH
Relative retention W ith reference to acetylsalicylic a d d
(retention tim e = about 5 min): impurity A = about 0.7;
im purity B = about 0.8; im purity C = about 1.3;
a OH
OH
— impurity B: not m ore than twice the area of the principal
peak in the chromatogram obtained with reference
solution (b) (0.2 p er cent);
— impurities F, G, I : for each impurity, not more than E. 2,2'- [(2-hydroxypropane-1,3-diyl)bis(oxy-4,1-phenylene)]
1.5 times the area of the principal peak in the diacetamide,
chrom atogram obtained with reference solution (b)
h 3c ch3
(0.15 p er cent); oh j
— unspecified impurities, for each impurity, not more th an the
area o f the principal peak in the chromatogram obtained
with reference solution (b) (0.10 per cent);
— totd: n o t more th an 5 limes the area of the principal peak F. 2,2'-[[(l-methylethyl)imino]bis[(2-hydroxypropane-3,l-
in the chrom atogram obtained with reference solution (b) diyI)oxy-4,1-phenylene]] diacetamide,
(0.5 per cent);
— disregard limit. 0.5 times the area o f the principal peak in V H H
^ ^ ch3
the chrom atogram obtained with reference solution (b) and enantiomer
(0.05 per cent). HO2C CH3
C h lo rid e s (2.4.4)
M axim um 0.1 per c e n t G . 2- [4- [(2i?5)-2-hydroxy-3- [(1 -methylethyl)amino]
Dissolve 50 mg in a mixture o f 1 m L o f dilute nitric acid R propoxy]phenyl] acetic a d d ,
and 15 m L o f water R. T he solution, without further addition
H OH
of dilute nitric acid R, complies with the test. H
N. .C H 3
L oss o n d ry in g (2.2.32) and enantiomer
D E F IN IT IO N
(3i?)-N-Methyl-3-(2-methylphenoxy)-3-phenylpropan-l-
B. 2-[4- [(2R5)-23-dihydroxypropoxy]phenyl] acetamide,
amine hydrochloride.
H OH C o n te n t
and enantiomer 98.0 per cent to 102.0 p er cent (dried substance).
CHARACTERS
D . 2-[4- [(2jR5)-3-chloro-2-hydroxypropoxy]phenyl] A p p e a ra n c e
acetamide, W hite or almost white powder.
S o lu b ility
Sparingly soluble in water, soluble in anhydrous ethanol,
practically insoluble in heptane.
1-208 Atomoxetine Hydrochloride 2016
It shows polymorphism (5.9). acid R previously adjusted to p H 2.5 w ith a 280 g/L solution
o f potassium hydroxide R
IDENTIFICATION
A. Infrared absorption spectrophotom etry (2.2.24). Test solution (a) Dissolve 25 m g o f the substance to be
examined in the mobile phase and dilute to 10.0 m L with
Comparison atomoxetine hydrochloride CRS.
the mobile phase.
If the spectra obtained in the solid state show differences,
Test solution (b) Dissolve 25.0 m g o f the substance to be
dissolve the substance to be examined and the reference
examined in the mobile phase and dilute to 100.0 m L with
substance separately in anhydrous ethanol R, evaporate to
the mobile phase.
dryness and record new spectra using the residues.
Reference solution (a) Dilute 1.0 m L o f test solution (a) to
B. Isomeric purity (see Tests).
100.0 m L with the mobile phase. D ilute 1.0 m L o f this
C. It gives reaction (a) o f chlorides (2.3.1). solution to 10.0 m L with the mobile phase.
TESTS Reference solution (b) Dissolve 7.5 m g o f 3-(methylammo)-l-
Isomeric purity phenylpropan-l-ol R (impurity H ) and 5 mg o f mandeHc add R
Liquid chromatography (2.2.29): use the normalisation (impurity E) in test solution (b) and dilute to 50 m L with
procedure. test solution (b).
Test solution Dissolve 35.0 m g of the substance to be Reference solution (c) Dissolve 5 m g o f atomoxetine for
examined in 2.5 m L o f anhydrous ethanol R, sonicate until impurity A identification CRS in the mobile phase and dilute
dissolution is complete and dilute to 10.0 m L with heptane R. to 20 m L with the mobile phase.
Reference solution (a) Dissolve 3.5 mg o f atomoxetine Reference solution (d) Dissolve 25.0 mg of atomoxetine
impurity B CRS and 1 mg o f atomoxetine impurity D CRS in hydrochloride CRS in the mobile phase and dilute to
5 m L of anhydrous ethanol R, sonicate until dissolution is 100.0 m L with the mobile phase.
complete and dilute to 20.0 m L with heptane R. Column:
Reference solution (b) Dissolve 35.0 m g of the substance to be — size. I = 0.15 m , 0 = 4.6 mm ;
examined in 2.5 m L o f anhydrous ethanol R. Add 1.0 m L of — stationary phase: end-capped octylsUyl silica gel for
reference solution (a) and dilute to 10.0 m L with heptane R. chromatography R (3.5 pm);
Reference solution (c) Dilute 1.0 m L of reference solution (a) — temperature. 40 °C.
to 100.0 m L with heptane R. Mobile phase propanol R, solution A (27:73 VIV).
Column: Flow rate 1.0 mL/min.
— size. I — 0.25 m , 0 = 4.6 mm; Detection Spectrophotom eter at 215 nm .
— stationary phase. cellulose derivative of silica gelfor chiral
Injection 10 pL of test solution (a) and reference solutions (a),
separation R (5 pm).
(b) and (c).
Mobile phase M ix 1.5 m L of diethylamine R, 2.0 m L of
Run time 2.5 times the retention time o f atomoxetine.
trifluoroacetic acid R and 150.0 m L o f 2-propanol R and dilute
to 1000 m l. with heptane R. Identification of impurities Use the chromatogram obtained
w ith reference solution (b) to identify the peaks due to
Flow rate 1.0 mL/min.
impurities E and H ; use the chromatogram supplied with
Detection Spectrophotom eter at 273 nm . atomoxetine for impurity A identification CRS and the
Irqecdon 10 pL of the test solution and reference solutions (b) chrom atogram obtained with reference solution (c) to identify
and (c). the peak due to impurity A.
Run time 1.3 times the retention time o f atomoxetine. Relative retention W ith reference to atomoxetine (retention
Identification of impurities U se the chromatogram obtained tim e = about 10 min): impurity E = about 0.2;
with reference solution (b) to identify the peaks due to impurity H = about 0.3; impurity A = about 0.7.
impurities B and D. System suitability: reference solution (b):
Relative retention W ith reference to atomoxetine (retention — resolution: m in im u m 5.0 between the peaks due to
time = about 12 min): impurity B = about 0.5; impurities E and H.
im purity D = about 0.6. Calculation ofpercentage contents:
System suitability: reference solution (b): — for each impurity, use the concentration o f atomoxetine
— resolution: minimum 1.8 between the peaks due to hydrochloride in reference solution (a).
impurities B and D . Limits:
Limits: — impurity A: maximum 0.3 per cent;
— impurity B: maximum 0.5 per cent; — unspecified impurities: for each impurity, maximum
— impurity D: maximum 0.15 per cent; 0.10 per cent;
— unspecified impurities', for each impurity, maximum — total: maximum 0.5 per cent;
0.10 per cent; — reporting threshold: 0.05 per cent.
— disregard limit: the area o f the peak due to impurity B in H e a v y m e ta ls (2.4.8)
the chromatogram obtained with reference solution (c) M axim um 10 ppm.
(0.05 per cent); disregard any peak with a relative Solvent mixture water R, methanol R (20:80 VIV).
retention with reference to atomoxetine o f about 0.7
0.250 g complies with test H. Prepare the reference solution
(impurity A).
using 0.25 m l . of lead standard solution (10 ppm Pb) R.
Related substances
L o ss o n d ry in g (2.2.32)
Liquid chromatography (2.2.29).
M axim um 0.5 per cent, determined on 1.000 g by drying
Solution A Dissolve 5.9 g of sodium octanesulfonate in vacuo at 105 °C for 2 h.
monohydrate R in 1000 m L o f a 2.9 g/L solution of phosphoric
2016 Atorvastatin Calcium Trihydrate 1-209
H . 3-(methylamino)-1 -phenylpropan-1-ol.
PhEur
A. N-methyl-3-phenoxy-3-phenylpropan-1-amine,
Atorvastatin Calcium Trihydrate ★ ★
★ ★
B . (3S)-N-methyl-3-(2-methylphenoxy)-3-phenylpropan-1-
amine,
HaC
DEFINITION
Calcium (3i?,5jR)-7-[2-(4-fluorophenyl)-5-(l-methyiethyl)-3-
phenyl-4-(phenylcarbamoyi)-1//-pyrrol- l-yQ-3,5-
dihydroxyheptanoate trihydrate.
H ,C
Content
97.0 p er cent to 102.0 p er cent (anhydrous substance).
D . (3R)-N-methyl-3-(3-methylphenoxy)-3-phenylpropan-1-
amine,
CHARACTERS
Appearance
W hite o r almost white powder.
HO H
Solubility
COjH
Very slightly soluble in water, slightly soluble in ethanol
(96 p er cent), practically insoluble in methylene chloride.
It shows polymorphism (5.9).
E. (2S)-2-hydroxy-2-phenylacetic acid (L-mandelic acid),
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24).
Comparison atorvastatin calcium trihydrate CRS.
1-210 Atorvastatin Calcium Trihydrate 2016
If the spectra obtained in the solid state show differences, impurity D CRS and 2.5 m g o f the substance to be examined
dissolve the substance to be examined and the reference in dimethylformamide R and dilute to 50.0 m L with the same
substance separately in methanol R, evaporate to dryness and solvent.
record new spectra using the residues. Column:
B. Enantiom eric purity (see Tests). — size: I = 0.25 m , 0 = 4.6 mm;
C. W ater (see Tests). — stationary phase: octylsifyl silica gel for chromatography R
(5 pm);
D. Ignite. T h e residue gives reaction (b) o f calcium (2.3.1).
— temperature: 35 °C.
Filtration may be necessary in case the residue does not
completely dissolve. Mobile phase:
— mobile phase A: tetrahydrofuran R, acetonitrüe R, 3.9 g/L
TESTS solution o f ammonium acetate R adjusted to p H 5.0 with
Enantiomeric purity glacial acetic acid R (12:21:67 VIVIV)',
Liquid chrom atography (2.2.29). — mobile phase B: tetrahydrofuran R, 3.9 g/L solution o f
Solvent mixture anhydrous ethanol R, methanol R (50:50 V!V). ammonium acetate R adjusted to p H 5.0 with glacial acetic
Test solution Dissolve 10 mg o f the substance to be examined add R, acetonitrüe R (12:27:61 VIVIV)',
in 4 m L o f the solvent mixture and dilute to 10.0 m L with
hexane R. Time Mobile phase A Mobile phase B
Reference solution (a) Dissolve 2 mg o f atorvastatin (min) (per cent V7V) (per cent V/V)
impurity E CRS in methanol R and dilute to 20.0 m L with the 0 • 40 100 0
same solvent (solution A). Dissolve 10 mg of the substance
40 - 70 100->20 0 + 80
to be examined in 1.25 m L of methanol R, add 0.75 m L of
solution A and 2 m L o f anhydrous ethanol R and dilute to 7 0 -8 5 20 0 80 -» 100
10.0 m L w ith hexane R.
Reference solution (b) T o 2.0 m L o f the test solution add Flow rate 1.5 mlVmin.
40.0 m L of the solvent mixture and dilute to 100.0 m L with
hexane R. T o 3.0 m L o f this solution add 5 m L of the Detection Spectrophotom eter at 244 nm .
solvent mixture and dilute to 20.0 m L with hexane R. Injection 20 )iL o f test solution (b) and reference solutions (b)
Column: and (c).
— size: I = 0.25 m, 0 = 4.6 mm; Identification of impurities U se the chrom atogram obtained
— stationary phase: amylose derivative of silica gel for with reference solution (c) to identify the peaks due to
chromatography R (10 jun). impurities A, B, C and D .
Mobile phase trifluoroacetic acid R, anhydrous ethanol R, Relative retention W ith reference to atorvastatin (retention
hexane R (0.1:6:94 VIVIV). time = about 33 min): impurity A = about 0.8;
Flow rate 1.0 mlVmin. im purity B = about 0.9; impurity C = about 1.2;
impurity D = about 2.1.
Detection Spectrophotom eter at 244 nm .
If necessary5 adjust the mobile phase by increasing or
Injection 20 )lL.
decreasing the percentage o f acetonitrile o r the p H o f the
Run time 1.2 times the retention time o f atorvastatin. am m onium acetate solution to achieve a retention time o f
Relative retention W ith reference to atorvastatin (retention about 33 rnin for atorvastatin. For example, raising the p H
time = about 44 min): impurity E = about 0.8. would decrease die retention time o f atorvastatin.
System suitability: reference solution (a): System suitability: reference solution (c):
— resolution: m inim um 2.0 between the peaks due to — resolution: m inim um 1.5 between the peaks due to
im purity E and atorvastatin. im purity B and atorvastatin.
Limit. Limits:
— impurity E: not m ore than the area o f the principal peak — impurities A , B: for each impurity, n o t m ore than 3 times
in the chrom atogram obtained with reference solution (b) the area o f the principal peak in the chrom atogram
(0.3 per cent). obtained with reference solution (b) (0.3 per cent);
Related substances — impurities C, D: for each impurity, n o t m ore than
Liquid chrom atography (2.2.29). 1.5 times die area o f the principal peak in the
chrom atogram obtained with reference solution (b)
Test solution (a) Dissolve 40.0 m g o f the substance to be
(0.15 p er cent);
examined in dimethytformamide R and dilute to 100.0 mT.
— unspecified impurities: for each im purity, not m ore than the
with the same solvent.
area o f the principal peak in the chrom atogram obtained
Test solution (b) Dissolve 50 m g o f the substance to be with reference solution (b) (0.10 p er cent);
examined in dimethylformamide R and dilute to 50.0 m L with — total: n o t m ore than 15 times the area o f the principal
the same solvent. peak in die chrom atogram obtained with reference
Reference solution (a) Dissolve 40.0 m g o f atorvastatin calcium solution (b) (1.5 p er cent);
trihydrate CRS in dimethylformamide R and dilute to — disregard limit. 0.5 tim es the area o f the principal peak in
100.0 m L with the same solvent. the chrom atogram obtained w ith reference solution (b)
Reference solution (b) D ilute 1.0 m L o f test solution (b) to (0.05 p er cent); disregard the peak due to
100.0 m L with dimethylformamide R. Dilute 1.0 m L o f this dimethylformamide.
solution to 10.0 m L w ith dimethylformarnide R. Sodium
Reference solution (c) Dissolve 2.5 mg o f atorvastatin M axim u m 0.4 per cent (anhydrous substance).
impurity A CRS, 2.5 m g o f atorvastatin impurity B CRS, Atomic absorption spectrometry (2.2.23, Method I).
2.5 m g o f atorvastatin impurity C CRS, 2.5 mg o f atorvastatin
2016 Atorvastatin Calcium Trihydrate 1-211
F . (3i?,5/$-7-[[(3i?,5£)-7-[2-(4-fluorophenyl)-5-
( 1-methylethyl)-3-phenyl-4-(phenylcarbamoyl)-1//-pyrrol-1-
yl] -3,5-dihydroxyheptanoyI] amino]-3,5-dihydroxyheptanoic
A. (3i?j5iR)-3j5-dihydroxy-7-[5-(l-methylethyl)-2j3-diphenyl-
add,
4-(phenylcarbamoyI)-1//-pyrrol-1-yl]heptanoic a d d
(desfluoroatorvastatin),
o
Test solution Dissolve 25.0 m g o f the substance to be
examined in the solvent mixture and dilute to 100.0 m L with
the solvent mixture.
Reference solution (a) Dissolve 25.0 m g o f atovaquone CRS in
the solvent mixture and dilute to 100.0 m L with the solvent
mixture.
Reference solution (b) Dissolve 2.5 m g o f atovaquone for system
suitability CRS (containing impurities B and C) in the solvent
mixture and dilute to 10.0 m L with the solvent mixture.
H. (4R,6R)-6-[2-[2-(4-fluorophenyI)-5-( 1-methylethyI)-3- Reference solution (c) Dilute 1.0 m L o f the test solution to
phenyl-4-(phenylcarbam oyl)-lif-pyrrol-l-yI]ethyl]-4- 100.0 m L with die solvent mixture. Dilute 1.0 m L o f this
hydroxytetrahydro-2Jï-pyran-2-one. solution to 10.0 m L with the solvent mixture.
__________________________________________________________ PhEur Column:
— size. I — 0.25 m, 0 = 4.6 mm;
— stationary phase, end-capped octadecylsifyl silica gelfor
chromatography R (5 pm).
Mobile phase phosphoric add R, methanol R2, water for
Atovaquone ****** chromatography R, acetomtrUe R1 (0.5:17.5:30:52.5 VIVIVIV).
(Ph E ut monograph 2192) * Flow rate 2.5 mL/min.
Detection Spectrophotometer at 220 nm.
Injection 20 jiL of the test solution and reference solutions (b)
and (c).
Run time Twice the retention time of atovaquone.
Identification of impurities Use the chromatogram supplied
O with atovaquone for system suitability CRS and the
chromatogram obtained with reference solution (b) to
C22H19C103 366.8 95233-18-4 identify the peaks due to impurities B and C.
Relative retention W ith reference to atovaquone (retention
Action and use time = about 15 min): impurity B = about 0.85;
Antiprotozoal (malaria). impurity C = about 0.90.
PhEur__________________________________________________________ System suitability: reference solution (b):
— resolution: minimum 2.0 between the peaks due to
DEFINITION impurity C and atovaquone;
2- [trans-4-(4-ChlorophenyI) cyclohexyl]-3- — peak-to-vaUey ratio: minim um 1.5, where Hp = height
hydroxynaphthalene-1,4-dione. above the baseline of the peak due to impurity C and
Content Hv ~ height above the baseline o f the lowest point of the
97.5 per cent to 102.0 per cent (anhydrous substance). curve separating this peak from the peak due to
impurity B.
CHARACTERS
Appearance Calculation of percentage contents:
Yellow, crystalline powder. — for each impurity, use the concentration o f atovaquone in
reference solution (c).
Solubility
Limits:
Practically insoluble in water, sparingly soluble in methylene
— impurity B: maximum 0.5 per cent;
chloride, very slightly soluble in methanol.
— impurity C: maximum 0.2 per cent;
It shows polymorphism (5.9). — unspecified impurities: for each impurity, maximum
IDENTIFICATION 0.10 per cent;
Infrared absorption spectrophotometry (2.2.24). — total: maximum 0.6 per cent;
Comparison atovaquone CRS. — reporting threshold: 0.05 per cent.
IM P U R IT IE S *****
Specified impurities B, C
Atracurium Besilate ★ ★
Other detectable impurities (the following substances would, if (Ph Ettr monograph 1970) *****
present at a sufficient level, be detected by one or other of
th e tests in the monograph. They are limited by die general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): A , D.
A ctio n a n d u se
Non-depolarizing neuromuscular blocker.
PhEir______________________________________
D E F IN IT IO N
B. 2 -[cw-4-(4-chlorophenyl)cyclohexyl]-3- Mixture of the cis-cis, cis-trans and trans-trans isomers of
hydroxynaphthalene-l,4-dione, 2 ,2 '- [pentane-1,5-diylbis [oxy(3-oxopropane-l ,3-diyl)]] bis [ 1-
(3,4-dimethoxybenzyl)-6,7-dimethoxy-2-methyl-l ,2,3,4-
tetrahydroisoquinolinium] dibenzenesulfonate.
C o n te n t
96.0 per cent to 102.0 per cent (anhydrous substance).
OH and enantiomer CHARACTERS
A p p e a ra n c e
White or yellowish-white, slightly hygroscopic powder.
C . 2- [( l.RS)-4-(4-chlorophenyl)cyclohex-3-en- 1-yl] -3- S olubility
hydroxynaphthalene-l,4-dione, Soluble in water, very soluble in acetonitrile, in ethanol
(96 per cent) and in methylene chloride.
ID E N T IF IC A T IO N
A Infrared absorption spectrophotometry (2.2.24).
Comparison atracurium besilate CRS.
B. Examine the chromatograms obtained in the assay.
Results T he 3 principal isomeric peaks in the chromatogram
D . 2-[mms-4-(4-chlorophenyl)cyclohexyl]-3- obtained with test solution (a) are similar in retention tim e to
methoxynaphthalene-1,4-dione. those in the chromatogram obtained with reference
solution (a).
PhEur
TESTS
S o lu tio n S
Dissolve 1.00 g in water R and dilute to 100 m L with the
same solvent.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and not more intensely coloured
than reference solution Y 7 (2.2.2, Method II).
R e la te d su b sta n c e s
Liquid chromatography (2.2.29).
Test solution (a) Dissolve 50.0 m g of the substance to be
examined in mobile phase A and dilute to 50.0 m L with
mobile phase A.
Test solution (b) Dissolve 0.100 g of the substance to be
examined in mobile phase A and dilute to 10.0 m L with
mobile phase A.
1-214 Atracurium Besilate 2016
Reference solution (a) Dissolve 50.0 m g o f atracurium im purity B = about 1.15; im purity C l = ab o u t 1.2;
besilate CRS in mobile phase A and dilute to 50.0 m L with impurity C2 (major isomer) = about 1.3.
mobile phase A. System suitability:
Reference solution (b) Dilute 1.0 m L o f test solution (a) to — resolution: minimum 1.5 betw een the peaks due to the
100.0 m L with mobile phase A. atracurium trans-trans isomer an d the atracurium cis-trans
Reference solution (c) Dissolve 20.0 m g o f methyl isomer, and m inim um 1.5 betw een the peaks due to the
benzenesulfonate R in aceîonitrüe R and dilute to 100.0 m l. atracurium cis-trans isom er and th e atracurium cis-ds
with the same solvent. Dilute 50 nL o f the solution to isomer in the chrom atogram obtained with reference
100.0 m L with mobile phase A. solution (a);
— pedk-to-valley ratio: m inim um 1.2, w here HP — height
Reference solution (d) Dissolve 2.0 m g o f atracurium for peak
above the baseline o f the peak due to im purity A l and
identification CRS (containing impurities A l, A2, B, C l , C2,
Hv = heigjit above the baseline o f the lowest point o f the
D l, D 2, E, G and K) in 2.0 m L o f mobile phase A.
curve separating this peak from th e peak due to the
Reference solution (e) Dissolve 2.0 m g o f atracurium for atracurium cis-cis isomer in the chrom atogram obtained
impurity F identification CRS in 2.0 m L o f mobile phase A. with reference solution (d).
Column: Limits:
— size: I = 0.25 m , 0 = 4.6 mm; — correction factor, for the calculation o f content, multiply the
— stationary phase: base-deactivated end-capped octadecylsifyl peak area of impurity G by 0.5;
silica gelfor chromatography R (5 |im ). — impurity E: not more than 1.5 tim es the sum o f the areas
Mobile phase: of the peaks due to the atracurium cis-cis, trans-trans and
— mobile phase A: mix 5 volumes of methanol R, 20 volumes cis-trans isomers in the chrom atogram obtained with
o f acetonitrüe R and 75 volumes o f a 10.2 g/L solution of reference solution (b) (1.5 per cent);
potassium dihydrogen phosphate R previously adjusted to — impurities A, D: for each im purity, for the sum o f the
p H 3.1 with phosphoric add R; areas o f the 2 isomer peaks, n o t m ore than 1.5 times the
— mobile phase B: mix 20 volumes o f acetomtrüe R, sum o f the areas o f the peaks due to the atracurium
30 volumes of methanol R and 50 volumes o f a 10.2 g/L cis-cis, trans-trans and cis-trans isomers in the
solution o f potassium dihydrogen phosphate R previously chromatogram obtained with reference solution (b)
adjusted to p H 3.1 with phosphoric add R; (1.5 per cent);
— impurity C: for the sum o f the areas o f the 2 isom er peaks,
Urne Mobile phase A Mobile phase B
not m ore than the sum o f the areas o f the peaks due to
(min) (per cent V/V) (percent V/V) the atracurium cis-cis, trans-trans and cis-trans isomers in
0 -5 80 20 the chrom atogram obtained w ith reference solution (b)
(1.0 p er cent);
5 -15 80 ->40 20 ->60
— impurities F, G: for each im purity, n o t m ore than the sum
15-25 40 60 o f the areas of the peaks due to th e atracurium cis-ds,
2 5 -3 0 40 ->0 60-> 100
trans-trans and cis-trans isomers in the chrom atogram
obtained with reference solution (b) (1.0 p er cent);
30 -4 5 0 100 — impurities H, 1, K. for the sum o f the areas o f the isom er
peaks o f these impurities, not m ore than th e sum o f the
areas o f the peaks due to the atracurium cis-cis, trans-trans
Flow rate 1 mlVmin.
and cis-trans isomers in the chrom atogram obtained with
Detection Spectrophotom eter at 280 run. reference solution (b) (1.0 per cent);
Injection 20 pL o f test solution (a) and reference solutions (a), — unspecified impurities: for each im purity, n o t m ore th an
(b), (d) and (e). 0.1 times the sum o f the areas o f the peaks due to the
Identification of impurities Use the chromatogram obtained atracurium cis-cis, trans-trans and cis-trans isomers in the
with reference solution (d) and the chromatogram supplied chromatogram obtained with reference solution (b)
with atracurium for peak identification CRS to identify the (0.10 p er cent);
peaks due to impurities A l, A2, B, C l, C2, D l, D 2, E, G — total: n o t more than 3.5 times th e sum o f the areas o f the
and K; use the chromatogram obtained with reference peaks due to the atracurium cis-cis, trans-trans and cis-trans
solution (e) and the chromatogram supplied with atracurium isomers in the chrom atogram obtained with reference
for impurity F identification CRS to identify the peak due to solution (b) (3.5 per cent);
im purity F. — disregard Umit. 0.05 times the sum o f the areas of the
Relative retention W ith reference to the atracurium tis-ds peaks due to the atracurium cis-cis, trans-trans an d cis-trans
isomer (retention time = about 30 min): isomers in the chrom atogram obtained with reference
impurity E = about 0.2; impurity F = about 0.25; solution (b) (0.05 per cent).
im purity G = about 0.3; impurity D l = about 0.45; Impurity J
impurity D 2 = about 0.5; atracurium Liquid chromatography (2.2.29) as described in the test for
trans-trans isom er = about 0.8; atracurium related substances with the following modifications.
cis-trans isomer = about 0.9; im purity A l = about 1.04;
im purity II = about 1.07; impurity H I = about 1.07
(shoulder on the front o f peak A2); im purity A2 (major
isomer) = about 1.08; impurity K1 = about 1.09 (shoulder
on the tail o f peak A2); impurity 12 (major
isomer) = about 1.12; impurity H 2 (major isomer) = about
1.12; impurity K 2 (major isomer) = about 1.12;
2016 Atracurium Besilate 1-215
Mobile phase:
5 - 15 8 0 -»75 20 ->25
15 - 25 75 25 o o
2 5 -3 0 75 55 25 ->45
3 0 -3 8 55 -» 0 45-» 100
38 - 45 0 100 A . l-(3,4-dimethoxybenzyl)-2-[13-[l-(3,4-dimethoxybenzyl)-
6,7-dim ethoxy-3,4-dihydroisoquinolin-2(l/i)-yI]-3,ll-dioxo-
4 ,1 0-dioxatridecyI]-6,7-dimethoxy-2-inethyl-l,2,3,4-
Detection Spectrophotom eter at 217 nm. tetrahydroisoquinolinium (A1 = cis-trans isomer,
A2 — cis-cis isomer),
Irtjection 100 jiL o f test solution (b) and reference
solution (c). o o
Retention time Im purity J = about 25 min; atracurium
trans-trans isomer = about 38 min. r a A q/ W ^ o^ r
Limit.
B. pentane-l,5-diyl bis[3-[l-(3,4-dimethoxybenzyl)-6,7-
— impurity J: n o t more than the area of the principal peak in
dimethoxy-3,4-dihydroisoquinolin-2( li/)-yl] prop anoate],
the chromatogram obtained with reference solution (c)
(10 ppm). o o
Isomer composition
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modifications. U se the
C . 1-(3,4-dimethoxybenzyl)-2-(3,11 -dioxo-4,10-dioxatridec-
normalisation procedure.
12-enyl)-6,7-dimethoxy-2-methyl-1,2,3,4-
Irtjection T est solution (a). tetrahydroisoquinolinium (C l = trans isomer,
Limits: C 2 = cis isomer),
— atracurium cis-cis isomer. 55.0 per cent to 60.0 per cent,
— atracurium cis-trans isomer. 34.5 per cent to 38.5 per cent, o
— atracurium trans-trans isomer. 5.0 per cent to 6.5 per cent.
Water (2.5.12)
M aximum 5.0 per cent, determined on 1.000 g. D . l-(3,4-dimethoxybenzyl)-2-[3-[(5-hydroxypentyl)oxy]-3-
Sulfated ash (2.4.14) oxopropyl] -6,7-dimethoxy-2-methyl-1,2,3,4-
M aximum 0.1 p er cent, determined on 1.0 g. tetrahydroisoquinolinium (D1 = trans isomer,
D 2 = cis isomer),
ASSAY
Liquid chromatography (2.2.29) as described in the test for . o
related substances with the following modification.
Irtjection T est solution (a) and reference solution (a).
Calculate the percentage content o f Q s H ^ ^ O j a S j from E . 2-(2-carboxyethyl)- l-(3,4-dimethoxybenzyl)-6,7-
the sum of the areas o f the peaks due to the 3 isomers. dimethoxy-2-methyl-1,2,3,4-tetrahydroisoquinolinium,
STORAGE F . R+-C H 3: l-(3,4-dimethoxybenzyl)-6,7-dimethoxy-2,2-
In an airtight containerj protected from light, at a dim ethyl-1,2,3,4-tetrahydroisoquinolinium,
tem perature o f 2 °C to 8 °C. G . R -C H 3: l-(3,4-dimethoxybenzyl)-6,7-dimethoxy-2-
IM P U R IT IE S m ethyl-1,2,3,4-tetrahydroisoquinoline,
Specified impurities A, C, D , E, F, G , H , I, J, K o
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the monograph. T hey are limited by die general O
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use H . 2,2 [hexane- 1,6-diylbis [oxy(3-oxopropane-1,3-
(2034). It is therefore n o t necessary to identify these dfyl)]]bis[l-(3,4-dimethoxybenzyl)-6,7-dimethoxy-2-methyl-
impurities for dem onstration of compliance. See also 5.10. I.2,3,4-tetrahydroisoquinolinium] (H I = cis-trans isomer,
Control of impurities in substances for pharmaceutical usé): B. H 2 = cis-cis isomer),
o o
Detecdon After cooling, spray with dilute potassium
’sV CHs iodobismuthate solution R.
Results T h e principal spot in the chrom atogram obtained with
the test solution is similar in position, colour and size to the
J. methyl benzenesulfonate, principal spot in the chrom atogram obtained with the
reference solution.
D . Place about 3 m g in a porcelain crudble and add 0.2 m L
of fuming nitric acid R. Evaporate to dryness on a water-bath.
Dissolve the residue in 0.5 m L o f a 30 g/L solution of
potassium hydroxide R in methanol R; a violet colour develops.
K. 2 ,2 '-[hexane-1j5-diylbis[oxy(3-oxopropane-l,3-diyl)]]
bis [ 1-(3 j4-dimethoxybenzyi)-6j 7-dimethoxy-2-methyl- E. Optical rotation (see Tests).
1,2,3,4-tetrahydroisoquinolinium]. TESTS
PhEur O p tic a l ro ta tio n (2.2.7)
—0.70° to + 0.05° (measured in a 2 dm tube).
Dissolve 1.25 g in ethanol (96 per cent) R and dilute to
25.0 m L with the same solvent.
★ ★ R e la te d su b s ta n c e s
Atropine ★ ★
Liquid chromatography (2.2.29).
*****
(Ph Eter monograph 2056) Test solution Dissolve 24 m g o f the substance to be examined
in mobile phase A and dilute to 100.0 m L with mobile
phase A.
Reference solution (a) D ilute 1.0 m L of the test solution to
and enanttomer
100.0 m L with mobile phase A. Dilute 1.0 m L o f this
solution to 10.0 m L with mobile phase A
Reference solution (b) Dissolve 5 mg o f atropine
impurity B CRS in the test solution and dilute to 20.0 m L
C j7H23N03 289.4 51-55-8 with the test solution. D ilute 5.0 m L o f this solution to
25.0 m L with mobile phase A.
A ctio n a n d use
Reference solution (c) Dissolve the contents o f a vial of atropine
Anticholinergic.
for peak identification CRS (containing impurities A, D , E, F,
PhEur. G and H ) in 1.0 m L o f mobile phase A
D E F IN IT IO N Reference solution (d) Dissolve 5 mg o f tropic acid R
(impurity C) in mobile phase A and dilute to 10.0 m L with
(1 i?,3i?,53)-8-Methyl-8-azabicyclo[3.2. l]oct-3-yl (2RS)-3-
hydroxy-2-phenylpropanoate. mobile phase A. Dilute 1.0 m L o f the solution to 100.0 m L
with mobile phase A. D ilute 1.0 m L o f this solution to
C o n te n t 10.0 m L with mobile phase A.
99.0 per cent to 101.0 p er cent (dried substance).
Column:
CHARACTERS — size. I = 0.10 m , 0 = 4.6 mm;
A p p e a ra n c e — stationary phase: octadecylsUyl silica gel for chromatography R
W hite or almost white, crystalline pow der or colourless (3 nm).
crystals. Mobile phase:
S o lu b ility — mobile phase A: dissolve 3.5 g o f sodium dodecyl sulfate R in
Very slightly soluble in water, freely soluble in ethanol 606 m L o f a 7.0 g/L solution o f potassium dihydrogen
(96 per cent) and in methylene chloride. phosphate R previously adjusted to p H 3.3 with a 5.8 g/L
solution o f phosphoric add R, and mix with 320 m L o f
ID E N T IF IC A T IO N
acetomtrüe R1;
First identification: A, B, E. — mobile phase B: acetomtrüe Rl;
Second identification: A , C, D, E
A. M elting point (2.2.14): 115 °C to 119 °C. Time Mobile phase A Mobile phase B
B. Infrared absorption spectrophotom etry (2.2.24). (min) (per cent V7V) (per cent V/V)
Comparison atropine CRS. 0 -2 95 5
STORAGE
Protected from light.
IM P U R IT IE S G. (lÄ,3r,5S)-8-methyl-8-azabicyclo[3.2.1]oct-3-yl (2RS)-2-
Specified impurities A, B, C , D , E, F , G, H . hydroxy-3-phenylpropanoate (littorine),
H . unknown structure.
PhEur
A. (12?,3r,55)-8-methyl-8-azabicydo[3.2.1]oct-3-yl
2-phenylpropenoate (apoatropine),
and enantiomer
Attapulgite
A c tio n a n d u se
A. (IR ,3r,55)-8-methyl-8-azabicydo[3.2.l]oct-3-yl Excipient.
2-phenylpropenoate (apoatropine)j
D E F IN IT IO N
Attapulgite is a purified native hydrated magnesium
alum inium silicate essentially consisting of the clay mineral
and enantiomer palygorskite.
C H A R A C T E R IS T IC S
A light, cream or buffi, very fine powder, free or almost free
from gritty particles.
B. (li?,3r,55)-8-azabicyclo[3.2.1]oct-3-yl (2RS)-3-hydroxy-2-
ID E N T IF IC A T IO N
phenylpropanoate (noratropine),
A. Ignite 0.5 g with 2 g of anhydrous sodium carbonate for
20 minutes, cool and extract with 25 m L o f boiling water.
Cool, filter, wash the residue with water and add the
CO2H washings to the filtrate. Reserve the residue for test B.
and enantiomer
Cautiously addify the combined filtrate and washings with
hydrochloric add, evaporate to dryness, moisten the residue
with 0.2 m l. of hydrochloric add, add 10 m l. of water and stir.
C . (2RS)-3-hydroxy-2-phenylpropanoic a d d (tropic ad d )j A white, gelatinous predpitate is produced.
B. W ash the residue reserved in test A with water and
H OH dissolve in 10 m L o f 2m hydrochloric add T o 2 m L o f the
solution add a 10% whr solution o f ammonium thiocyanate.
« ^ /•4 — r H
and epimer at C* A n intense red colour is produced.
, v -o -\
C. T o 2 m L of the solution obtained in test B add 1 m L of
\ h I strong sodium hydroxide solution and filter. T o the filtrate add
3 m L of ammonium chloride solution. A gelatinous white
D . (IR,3S,5R,6RS)- 6-hydroxy-8-methyl-8 - predpitate is produced.
a2abicydo[ 3 .2 . l]oct-3-yl (25)-3-hydroxy-2-phenylpropanoate D . T o 2 m L o f the solution obtained in test B add
( 6-hydiroxyhyoscyamine), ammonium chloride and an excess o f 13.5m ammonia and
1-220 Attapulgite 2016
filter. T o the filtrate add 0.15 m L o f magneson reagent and an Loss on ignition
excess o f 5 m sodium hydroxide. A blue precipitate is produced. W hen ignited at 600°, loses 15.0 to 27.0% o f its weight.
Use 1 g.
TESTS
Acidity or alkalinity
p H o f a 5% wN suspension in carbon dioxide-free water, after
shaking for 5 minutes, 7.0 to 9.5, Appendix V L.
Adsorptive capacity Activated Attapulgite
M oisture adsorption, 5 to 14% when determined by the
following method. D ry in air and pow der a sufficient quantity Action and use
o f th e substance being examined and pass through a sieve Antidiarrhoeal. '
with a nominal m esh aperture of 150 jam. Spread 0.5 g as a
DEFINITION
thin layer on a previously weighed piece o f aluminium foil
Activated Attapulgite is a purified native hydrated magnesium
(60 m m x 50 mm ) o f nominal gauge 17.5 nm and transfer
alum inium silicate essentially consisting o f the clay mineral
to a desiccator containing a dish o f sodium chloride crystals
palygorskite that has been carefully heated to increase its
partially immersed in saturated brine at 25°. After 4 hours,
adsorptive capacity.
remove from the desiccator and weigh immediately. D ry in
an oven at 110° for 4 hours, allow to cool in a desiccator and CHARACTERISTICS
weigh. T he moisture adsorption is the gain in weight o f the A light, cream or buff, very fine powder, free or almost free
substance being examined expressed as a percentage o f its from gritty particles.
oven-dried w eight
IDENTIFICATION
Arsenic A. Ignite 0 .5 g with 2 g o f anhydrous sodium carbonate for
T o 0.13 g add 5 m L o f water, 2 m L o f sulfuric add and 2 0 minutes, cool and extract with 2 5 m L o f boiling water.
10 m L of sulfur dioxide solution and evaporate on a water bath Cool, filter, wash the residue with water and add the
until the sulfur dioxide solution is removed and the volume washings to the filtrate. Reserve the residue for test B.
reduced to about 2 mL. Transfer the solution to the Cautiously acidify the combined filtrate and washings with
generator flask with the aid o f 5 m L o f water. T he resulting hydrochloric add, evaporate to dryness, m oisten the residue
solution complies with the limit test for arsenic, Appendix VII with 0 .2 m L o f hydrochloric add, add 10 m L of water and stir.
(8 ppm ). A white, gelatinous precipitate is produced.
Heavy metals B. W ash the residue reserved in test A with water and
A. N o t m ore than 2 0 ppm when determined by the following dissolve in 10 m L o f 2 m hydrochloric add. T o 2 m L of the
m ethod. Shake 6 .0 g with 4 0 m L o f 0 .5 m hydrochloric add at solution add a 10% w/v solution of ammonium thiocyanate.
3 7 ° for 3 0 minutes, cool and filter. W ash the residue with A n intense red colour is produced.
water and dilute the combined filtrate and washings to 5 0 m L C. T o 2 m l, o f the solution obtained in test B add 1 m L of
with water. T o 2 0 m L add 2 g each o f ammonium chloride and strong sodium hydroxide solution and filter. T o the filtrate add
ammonium tkiocyanate and dissolve. Shake the solution with 3 m L of ammonium chloride sdution. A gelatinous white
8 0 m L of a mixture o f equal volumes o f ether and isoamyl precipitate is produced.
alcohol and separate, retaining the aqueous layer. Extract with
D . T o 2 m L o f the solution obtained in test B add
a further 8 0 m L of the mixture. T o the aqueous layer add
ammonium chloride and an excess o f 1 3 .5 m ammonia and
2 g o f citric add, neutralise with 1 3 .5 m ammonia and dilute to
filter. T o the filtrate add 0 .1 5 m L o f magneson reagent and an
25 m L with water. 12 m L o f the resulting solution complies
excess o f 5 m sodium hydroxide. A blue precipitate is produced.
with limit test A for heavy metals, Appendix VII. Use lead
standard solution (2 ppm Pb) to prepare the standard. TESTS
B. N o t more than 10 ppm when determined by the following Acidity or alkalinity
m ethod. Shake 6.0 g with 40 m L o f 0.5m sodium hydroxide at p H o f a 5% w/v suspension in carbon dioxide-free water, after
37° for 30 minutes, cool and filter. W ash the residue with shaking for 5 minutes, 7 .0 to 9 .5 , Appendix V L.
water and dilute the combined filtrate and washings to 50 mT. Arsenic
with water. Neutralise 20 m L o f the solution with hydrochloric T o 0 .1 3 g add 5 mT, o f water, 2 m L o f sulfuric add and
add and dilute to 25 m L with water. 12 m L o f the resulting 10 mT, o f sulfur dioxide solution and evaporate on a w ater bath
solution complies with limit test A for heavy metals, until the sulfur dioxide solution is removed and the volume
A ppendix VH. Use lead standard solution (1 ppm Pb) to reduced to about 2 mT. Transfer the solution to the
prepare the standard. generator flask with the aid o f 5 m L o f water. T h e resulting
Acid-soluble m atter solution complies with the limit test for arsenic, Appendix VII
Boil 2 g with 1 0 0 m L o f 0 .2 m hydrochloric add under a reflux (8 ppm).
condenser for 5 minutes, cool and filter. Evaporate 5 0 mT, o f Heavy m etals
the filtrate to dryness. T he residue, after ignition at about A. N o t m ore than 2 0 ppm when determ ined by the following
6 0 0 ° for 3 0 minutes, weighs not m ore than 0 .2 5 g. m ethod. Shake 6 .0 g with 4 0 m L of 0.5m hydrochloric add at
W ater-soluble matter 3 7 ° for 3 0 minutes, cool and filter. W ash the residue with
Boil 10 g with 100 m L o f water under a reflux condenser for water and dilute die combined filtrate and washings to 5 0 m L
5 m inutes, cool and filter. Evaporate 50 m L o f the filtrate to with water. T o 2 0 mT, add 2 g each of ammonium chloride and
diyness. T h e residue, after ignition a t 600° for 30 minutes, ammonium thiocyanate and dissolve. Shake the solution with
weighs n o t m ore than 50 mg. 8 0 m L o f a mixture o f equal volumes o f ether and isoamyl
alcohol and separate, retaining the aqueous layer. Extract with
Loss on drying
a further 8 0 m L o f the mixture. T o the aqueous layer add
W hen dried to constant weight at 105°, loses n o t more than
2 g o f citric add, neutralise with 13.5m ammonia and dilute to
17.0% o f its weight. Use 1 g.
2 5 m L w ith water. 12 m L o f the resulting solution complies
2016 Azapropazone 1-221
with limit test A for heavy metals, Appendix VII. Use lead IDENTIFICATION
standard solution (2 ppm Pb) to prepare die standard. A. T he infrared absorption spectrum, Appendix II A, is
B. N o t m ore than 10 ppm w hen determined by the following concordant with die reference spectrum of azapropazone
method. Shake 6.0 g with 40 m L o f 0.5m sodium hydroxide at (RS 016).
37° for 30 m in u tes, cool and filter. W ash die residue with B. T he light absorption, Appendix I I B , in the range 230 to
water and dilute the combined filtrate and washings to 50 m L 350 nm of a 0.0008% w/v solution in 0.1 m sodium hydroxide
with water. Neutralise 20 m L o f the solution with hydrochloric exhibits two maxima, at 255 nm and 325 nm . T he
acid and dilute to 25 m L with water. 12 m L of the resulting absorbances at 255 nm and 325 nm are about 0.86 and 0.18
solution complies with limit test A for heavy metals, respectively.
Appendix VII. U se lead standard solution (1 ppm Pb) to
TESTS
prepare the standard.
Acetic add
A d d -s o lu b le m a t te r N o t more than 0.2%, determined by the following m ethod.
Boil 2 g with 100 m L o f 0.2m hydrochloric acid under a reflux Dissolve 10 g in 25 m L o f methanol, add 75 m L of water and
condenser for 5 m inutes, cool and filter. Evaporate 50 m L of carry out a potendometric titration, Appendix V H IB , using
the filtrate to dryness. T h e residue, after ignition at about 0.1 m sodium hydroxide KS as titrant to a p H o f 5.9. Each m L
600° for 30 minutes, weighs n o t more than 0.25 g. o f 0.1 m sodium hydroxide FS is equivalent to 6.005 mg of
W ate r-so lu b le m a t te r acetic ad d , C 2H 4O 2.
Boil 10 g with 100 m L of water under a reflux condenser for Related substances
5 minutes, cool and filter. Evaporate 50 m L of the filtrate to Carry out the following operations in subdued light using
dryness. T h e residue, after ignition at 600° for 30 minutes, low-actinic glassware without delay. Carry out the method
weighs not more than 50 mg. for liquid chromatography, Appendix HI D , using the following
A d so rp tiv e c a p a c ity solutions in a mixture of 1 volume of phosphate buffer pH 4.0
In a stoppered bottle shake 1.0 g, in very fine powder, with and 3 volumes of methanol.
50 m L o f a 0.12% w/v solution of methylene blue for ( 1) 0 . 10% w/v of die substance being examined.
5 minutes, allow to setde and centrifuge. T h e colour o f the (2) 0.00010% w/v o f azapropazone impurity A BPCRS.
clear supernatant solution is no t more intense than that of a
(3) 0.00025% w/v of azapropazone impurity B BPCRS.
0.0012% w/v solution of methylene blue.
(4) 0.00025% w/v of azapropazone impurity C BPCRS.
L oss o n d ry in g
W hen dried to constant weight at 105°, loses not more than (5) 0.0001% w/v o f the substance being examined.
4.0% of its weight. U se 1 g. ( 6) 0.00005% w/v of the substance being examined.
L oss o n ig n itio n (7) 0.1% w/v o f azapropazone impurity standard BPCRS.
W hen ignited at 600°, loses n o t more than 9.0% of its CHROMATOGRAPHIC CONDITIONS
weight. U se 1 g. (a) U se a stainless steel column (30 cm x 3.9 mm) packed
with octadecylsUyl silica gel for chromatography (10 nm)
(jiBondapak C18 is suitable).
(b) U se isocratic elution and the mobile phase described
Azapropazone below.
(c) U se a flow rate of 2.5 m L per minute.
(d) U se ambient column temperature.
(e) Use a detection wavelength of 254 nm.
(f) Inject 20 |iL o f each solution.
M OBILE PHASE
1 volume o f glacial acetic acid, 36 volumes of methanol and
63 volumes o f a 0.068% w/v solution of sodium
Ci6H 2oN40 2,2H20 336.4 13539-59-8 (anhydrous) butanesulfonate in water.
SYSTEM SUITABILITY
A ction a n d u se
Inject solution (7) and continue the chromatography for
Cyclo-oxygenase inhibitor; analgesic; anti-inflammatory.
5 times the retention time of the principal peak.
P re p a r a tio n s
T h e test is n o t valid unless the chromatogram obtained with
Azapropazone Capsules
solution (7) closely resembles the reference chromatogram
Azapropazone Tablets supplied with the azapropazone impurity standard.
I f necessary adjust the proportion o f methanol in the mobile
D E F IN IT IO N
phase to give the required retention times.
Azapropazone is 5-dimethylamino-9-methyl-2-
LIMITS
propylpyrazolo [ 1,2-a] [1,2,4] benzotriazine-1,3 (2 H) -dione
dihydrate. It contains not less than 99.0% and n o t m ore than In the chromatogram obtained w ith solution (1):
101.0% of C 16H 2oN402 , calculated with’reference to the the area of any peak corresponding to azapropazone
anhydrous substance. im purity A is n o t greater than the area o f the corresponding
C H A R A C T E R IS T IC S peak in the chromatogram obtained with solution (2) (0 . 1%);
A white to pale yellow, crystalline powder. the area of any peak corresponding to azapropazone
Very slightly soluble in water, soluble in ethanol (96%)-, im purity B is n o t greater than the area of the corresponding
it dissolves in solutions of alkali hydroxides..
1-222 Azathioprine 2016
Disregard any peak with an area less than the area o f the
principal peak in the chromatogram obtained with solution
(6) (0.05%). 4. a-(3-dimethylamino-7-methyl-1,2-dihydro-1,2,4-
H eav y m e ta ls benzotriazin-2-ylcaibonyl)valeric a d d (impurity C).
Dissolve 3.125 g in 20 m L o f water, boil for 10 minutes,
dilute to 50 m L and filter (solution A). 12 m L o f solution A
complies with limit test A for heavy metals, Appendix VII.
Use 10 m L o f lead standard solution (1 ppm Pb) to prepare the ★.★ ** ★
standard (16 ppm). Azathioprine ★ ★
*****
C h lo rid e (Ph. Eur. monograph 0369)
A m ixture o f 10 m L o f solution A and 5 m L o f water
complies with the limit test for chlorides, Appendix VII ,n o 2
n -—y
(80 ppm ).
S u lfate
<N x S
A mixture o f 10 m L o f solution A and 5 m L o f water
complies with the limit test for sulfates, Appendix VII
(240 ppm ).
cx>
W a te r
10.0 to 11.5% w/w, Appendix IX C. Use 0.25 g. C9H7N70 2S 277.3 446-86-6
S u lfa te d a s h
A ctio n a n d u se
N o t m ore than 0.1%, Appendix IX A
Immunosuppressant.
A SSA Y P re p a ra tio n s
Carry out M ethod I for non-aqueous titration, Azathioprine Tablets
A ppendix V III A, using 0.25 g and determining the end
Azathioprine Oral Suspension
point potentiometrically. E ach m L o f 0 .1 m perchloric acid VS
is equivalent to 30.04 mg o f C 16H 2oN40 2. PhEur__________________________
IM P U R IT IE S D E F IN IT IO N
6- [( l-M ethyl-4-nitro- lH-imidazol-5-yl)sulfenyl] -7H-purine.
Me N. C o n te n t
'N
98.5 per cent to 101.0 per cent (dried substance).
X
N NM62 CHARACTERS
A p p e a ra n c e
1. 3-dimethylamino-7-methyi-l ,2,4-benzotriazine Pale-yellow powder.
(impurity A ), S o lubility
Practically insoluble in water and in ethanol (96 p er cent).
Bun It is soluble in dilute solutions of alkali hydroxides and
sparingly soluble in dilute mineral adds.
Me H L
ID E N T IF IC A T IO N
N
A NMe2 Infrared absorption spectrophotom etry (2.2.24).
Comparison azathioprine CRS.
TESTS
2. 3-dimethylamino- l,2-dihydro-7-methyl-2-valeryl-1,2,4-
R e la te d su b s ta n c e s
benzotriazine,
Liquid chromatography (2.2.29).
Solution A 2.76 g/L solution o f sodium dihydrogen phosphate
monohydrate R adjusted to p H 2.5 with phosphoric acid R.
Test solution Dissolve 10 m g o f the substance to be examined
in 35 m L o f a 0.8 g/L solution o f sodium hydroxide R and
dilute to 100.0 m L with solution A.
2016 Azathioprine 1-223
SH
<x
no 2
reference solution (b) to identify the peak due to impurity G.
Relative retention W ith reference to azathioprine (retention N ^C I
tim e = about 15 min): impurity A = about 0.3; H3C
im purity B = about 0.4; impurity G = about 0.97.
System suitability: C. 5-chloro-l-methyl-4-nitro- l//-im idazole,
— resolution: minimum 2.0 between the peaks due to
impurities A and B in the chromatogram obtained with NOj
reference solution (a); minimum 2.0 between the peaks n —/
due to impurity G and azathioprine in the chromatogram <N x
-^ S H
obtained with reference solution (b).
H3C
Limits:
— impurities A, B: for each impurity, not more than
1.5 times the area o f the principal peak in the D . 1-methyl-4-nitro- li/-imidazole-5-thiol,
chromatogram obtained with reference solution (c)
(0.15 per cent); ,NOj
N—/
— unspecified impurities: for each impurity, not m ore than the
area of the principal peak in the chromatogram obtained
<N 1 OH
with reference solution (c) (0.10 per cent); H,C
— total: n o t more than 5 times the area of the principal peak
in the chromatogram obtained with reference solution (c) E. l-methyl-4-nitro-lH-imidazol-5-ol,
(0.5 per cent);
— disregard Hmir. 0.5 times the area o f the principal peak in
the chromatogram obtained with reference solution (c)
(0.05 per cent).
L o ss o n d ry in g (2.2.32)
M axim um 1.0 per cent, determined on 0.500 g by drying in
an oven at 105 °C. F. 1,7-dihydro-6//-purin-6-one (hypoxanthine),
1-224 Azelastine Hydrochloride 2016
Hj N
xX> 100.0 m L w ith the solvent mixture. D ilute 1.0 m L o f this
solution to 10.0 m L with the solvent mixture.
Reference solution (b) Dissolve 1 m g o f azdastine
G . 6- [(1 -m ethyl-4-nitro-1//-imidazol-5-yl) sulfanyl] -7//-p u rin- impurity B CRS, 1 m g o f azdastine impurity D CRS and 1 mg
2-amine (thiamiprine). o f azdastine impurity E CRS in the test solution and dilute to
PhEur 20 m L w ith the test solution.
Column:
— sizer. I = 0.25 m , 0 = 4.6 m m ,
— stationary phase: nitrUe silica gd for chromatography R
★ ★ (10 pm ),
Azelastine Hydrochloride ★ ★ — temperature: 30°C.
(Ph Eur monograph 1633) ***** Mobile phase Dissolve 2.16 g o f sodium octanesulfonate R and
0.68 g o f potassium dihydrogen phosphate R in 740 m L o f water
for chromatography R, adjust to p H 3.0-3.1 w ith dilute
phosphoric add R, add 260 m L o f acetonitrile for
chromatography R and mix.
, Ha Flow rate 2.0 m lA nin.
Detection Spectrophotom eter at 210 nm .
Irgecdon 10 |iL.
Run time Twice the retention tim e of azelastine.
Relative retention W ith reference to azelastine (retention
C 22H 25Q 2N 3O 418.4 79307-93-0 tim e = about 8-9 min): im purity A = about 0.2;
impurity B = about 0.3; im purity C = about 0.4;
A ctio n a n d u se
impurity D = about 0 .6; im purity E = about 1.4.
Histamine H x receptor antagonist; antihistamine.
System suitability: reference solution (b):
PhEir. — resolution: m inim um 4.0 between the peaks due to
impurities B and D ,
D E F IN IT IO N
— the peaks due to impurities D and E are baseline
4-(4-Chlorobenzyi)-2- [(4i25)-l-methylhexahydro- lH -azepin-
separated from the principal peak.
4-yI]phthalazin-1 (2H)-onc hydrochloride.
Limits:
C o n te n t — correction factors: for the calculation o f content, multiply
99.0 per cent to 101.0 p er cent (dried substance). the peak areas o f the following impurities by the
CHARACTERS corresponding correction factor, im purity B = 3.6;
A p p e a ra n c e impurity D = 0.7; im purity E = 2 . 1;
W hite or alm ost white, crystalline powder. — impurities A , B, C, D, E: for each impurity, n o t m ore than
the area of the principal peak in the chrom atogram
S o lu b ility
obtained with reference solution (a) (0.1 p er cent);
Sparingly soluble in water, soluble in ethanol and in
— any other impurity: for each impurity, n o t m ore than the
methylene chloride.
area o f th e principal peak in the chrom atogram obtained
ID E N T IF IC A T IO N with reference solution (a) (0.1 per cent);
A Infrared absorption spectrophotometry (2.2.24). — totah n o t m ore than twice the area o f the principal peak in
Comparison azdastine hydrochloride CRS. the chrom atogram obtained with reference solution (a)
B. Solution S (see Tests) gives reaction (a) o f chlorides (0.2 per cent);
(2.3.1). — disregard limit. 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (a)
TESTS (0.05 p er cent).
S o lu tio n S
L o ss o n d ry in g (2.2.32)
Dissolve 1.0 g in carbon dioxide-free water R and dilute to
M axim um 0.5 p e r cent, determ ined on 1.000 g by drying in
100 m L w ith the same solvent.
an oven at 105 °C.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
ASSAY
In order to avoid overheating in the reaction medium, mix
A cid ity o r a lk a lin ity thoroughly throughout and stop the titration immediately after the
T o 10 m L o f solution S add 0.2 m L o f bromothymol blue end-point has been reached.
solution R l. N o t m ore than 0.1 m L o f 0.01 M hydrochloric
Dissolve 0.300 g in 5 m L o f anhydrous formic add R.
acid or 0.01 M sodium hydroxide is required to c h a n g e the
A dd 30 m L o f acetic anhydride R. T itrate quickly w ith 0.1 M
colour of the solution.
perchloric acid, d eterm in in g the end-point potentiom etrically
R e la te d su b s ta n c e s (2. 2. 20).
Liquid chrom atography (2.2.29).
1.0 m L o f 0.1 M perchloric add is equivalent to 41.84 m g o f
Solvent mixture acetomtrUe for chromatography R, water R C 22H 25CI2N 3O.
(45:55 V/V).
2016 Azithromycin 1-225
IM P U R IT IE S ★>*★★
Azithromycin * ★
Specified impurities: A , B, C, D, E.
(Ph. Eur. monograph 1649) *****
ch 3
NH
i
nh2
A. benzoyldiazane (benzohydrazide),
and enantiomer
sCH3
0
A ctio n aind u se
Macrolide antibacterial.
PhEur_____________________________________
C. 2-[(4-chlorophenyl)acetyl]benzoic add, D E F IN IT IO N
(2R,3S,4R,5R,8R, 10R, 11R, 125,135,14R)-13- [(2,6-Dideoxy-
o 3-C-methyl-3-0-methyl-oc-L-nfco-hexopyranosyl) oxy] -2-ethyl-
3,4,1O-trihydroxy-3,5,6,8, 10,12,14-heptamethyl-l 1-[[3,4,6-
trideoxy-3- (dimethylamino)-P-D-ry/o-hexopyranosyl] oxy] -1 -
oxa-6-azacydopentadecan-15-one. T he degree of hydration is
1 or 2.
Semi-synthetic product derived from a fermentation p roduct
C o n te n t
96.0 per cent to 102.0 per cent (anhydrous substance).
D. 4-(4-chlorobenzyl)phthalazm-l (2/i)-one, CHARACTERS
A p p e a ra n c e
0 W hite or almost white powder.
S olu b ility
o
Practically insoluble in water, freely soluble in anhydrous
ethanol and in methylene chloride.
ID E N T IF IC A T IO N
Infrared absorption spectrophotometry (2.2.24).
Comparison azithromycin CRS.
If the spectra obtained in the solid state show differences,
prepare further spectra using 90 g/L solutions in methylene
E. 3-(4-chlorobenzylidene)isobenzofuran-l(3if)-one. chloride R.
PhEur TESTS
S o lu tio n S
Dissolve 0.500 g in anhydrous ethanol R and dilute to
50.0 m L with the same solvent.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
p H (2.2.3)
9.0 to 11.0.
Dissolve 0.100 g in 25.0 m L of methanol R and dilute to
50.0 m L with carbon dioxide-free water R.
Specific o p tic a l ro ta tio n (2.2.7)
—45 to —49 (anhydrous substance), determined on
solution S.
R e la te d su b stan c es
Liquid chromatography (2.2.29).
1-226 Azithromycin 2016
ch3
Mobile phase Mix 60 volumes of acetomtrüe R1 and h 3c /— u ch3
40 volumes o f a 6.7 g/L solution o f dipotassium hydrogen
phosphate R adjusted to p H 11.0 with a 560 g/L solution of
potassium hydroxide R
Flow rate 1.0 mL/min.
ch3
Detection Spectrophotometer at 210 nm .
cladinosyl
Injection 10 |xL.
h ch3
Run time 1.5 times the retention time of azithromycin.
Retention time Azithromycin = about 10 min.
System suitability: reference solution (b):
— resolution: minimum 3.0 between the peaks due to
im purity A and azithromycin. R1 = dadinosyl R2 = ( NH2
R1 = H R2 =
J. 13-O-decladinosylazithromycin,
A. R1 = O H , R2 = R6 = H , R3 = R4 = R5 = C H 3: ch3
6-dem ethylazithromyán,
H c / o
B. R1 = R 6 = H , R2 = R3 = R4 = R5 = C H 3: R1 = dadinosyl R2 = 3 ^ n —c h
3-deoxyazithromydn (azithromycin B),
C. R1 = O H , R2 = R3 = R5 = C H 3, R4 = R6 = H : 3 "-0 - OH
dem ethylazithrom ydn (azithromycin C),
D. L. azithromycin 3 '-iV-oxide,
R1 = O H , R2 = R3 = R4 = CH3, R5 = C H 2O H , R6 = H:
14-dem ethyl-14- (hydroxymethyl) azithromycin (azithromycin
0 f»
F ),
F. R1 = O H , R2 = R4 = R5 = C H 3, R3 = C H O , R6 = H: R1 = dadinosyl R2 = HM r
C NH
3 '-N -dem ethyl-3 '-AT-foimylazithromycin,
I. R1 = O H , R2 = R4 = R5 = C H 3, R3 = R6 = H: 3 '-N- OH
demethylazithromycin,
O. R1 = O H , R2 = R3 = R4 = R5 = R6 = C H 3: M . 3 '-(N,N-didem ethyl)-3 '-N’-foimylazithromycin,
2-desethyl-2-propylazithromycin,
ch3
R1 * cladinosyl R2 =
O ÓH
N . 3 '-de(dime&ylamino)-3 '-oxoazithromycin,
1-228 Bacampicillin Hydrochloride 2016
ch3
Mobile phase M ix 10 volumes o f a 272 g/L solution o f sodium
acetate R adjusted to p H 5.0 with glacial acetic add R}
40 volumes o f water R and 50 volumes o f ethanol
(96 per cent) R.
Application 1 pL.
Development Over a p ath o f 15 cm.
Drying In a current o f warm air.
Detection Spray with nmhydrin solution R1 and heat a t 60 °C
for 10 min.
System suitability, reference solution (b):
— the chrom atogram shows 3 clearly separated spots.
K. C 14j 1 "-epoxyazithromydn (azithromycin E), Results T h e principal spot in the chrom atogram obtained with
the test solution is similar in position, colour and size to the
P. unknow n structure.
principal spot in the chromatogram obtained with reference
PhEur solution (a).
C. Place about 2 m g in a test-tube about 150 mm long and
15 m m in diameter. M oisten with 0.05 m L o f water R and
add 2 m L o f sulfuric add-formaldehyde reagent R. M ix the
Bacampicillin Hydrochloride ★ ★ contents of the tube by swirling; the solution is practically
★ ★
colourless. Place the test-tube on a water-bath for 1 min;
(Ph. Eur. monograph 0808) * * * * *
a dark yellow colour develops.
D . Dissolve about 25 m g in 2 m L o f water R. A dd 2 m L of
dilute sodium hydroxide solution R and shake. W ait a few
h | V H3i
m inutes and add 3 m L o f dilute nitric add R and 0.5 m l. o f
H NH Y n^ A ^ ch ,
H'. / NH2h O V c h 3 silver nitrate sohaion R l. A white precipitate is formed.
A dd 0.5 m l. o f concentrated ammonia R T h e precipitate
H H dissolves.
® and epimer at C*
TESTS
A p p e a ra n c e o f so lu tio n
C21H28CIN3O7S 502.0 37661-08-8
Dissolve 0.200 g in 20 m L o f water R; the solution is not
A c tio n a n d u se m ore opalescent than reference suspension II (2.2.1).
Penicillin antibacterial. Dissolve 0.500 g in 10 m L o f water R; the absorbance
(2.2.25) o f the solution at 430 nm is n o t greater th an 0.10.
PhEur. p H (2.2.3)
D E F IN IT IO N 3.0 to 4.5.
( IRS ) - 1-[(Ethoxycarbonyl)oxy]ethyl (2S,5R,6R)-6-[[(2R)-2- Dissolve 1.0 g in carbon dioxide-free water R and dilute to
amino-2-phenylacetyl] amino] -3 ,3-dime thyl-7 -oxo-4-thia-1 - 50 m L with the same solvent.
azabicyclo [3.2.0]heptane-2-carboxylate hydrochloride. S p ecific o p tic a l ro ta tio n (2.2.7)
Semi-synthetic product derived from a fermentation product. + 175 to + 195 (anhydrous substance).
C o n te n t Dissolve 0.250 g in w ater R and dilute to 25.0 m L with the
95.0 p er cent to 102.0 per cent (anhydrous substance). same solvent.
CHARACTERS R e la te d su b s ta n c e s
A p p e a ra n c e L iquid chrom atography (2.2.29). Prepare the test solution and
W hite or alm ost white pow der or granules, hygroscopic. reference solutions (a), (b) and (d) immediately before use.
S o lu b ility Phosphate buffer A Dissolve 1.4 g of sodium dihydrogen
Soluble in water, freely soluble in ethanol (96 per cent), phosphate monohydrate R in water R and dilute to about
soluble in methylene chloride. 800 m l. with the same solvent. Adjust to p H 3.0 w ith dilute
phosphoric add R and dilute to 1000.0 m L with water R.
ID E N T IF IC A T IO N
Phosphate buffer B Dissolve 2.75 g o f sodium dihydrogen
First identification A , D phosphate monohydrate R and 2.3 g o f disodium hydrogen
Second identification B, C, D phosphate dihydrate R in water R and dilute to about 1800 m L
A Infrared absorption spectrophotom etry (2.2.24). with the sam e solvent. Adjust to p H 6.8, if necessary, using
Comparison bacampidUin hydrochloride CRS. dilute phosphoric add R or dilute sodium hydroxide solution R
and dilute to 2000.0 m L w ith water R.
B. Thin-layer chromatography (2.2.27).
Test solution Dissolve 10 mg o f the substance to be examined Test solution Dissolve 30.0 m g o f the substance to be
in 2 m L o f methanol R. examined in phosphate buffer A and dilute to 100.0 m L with
phosphate buffer A.
Reference solution (a) Dissolve 10 m g o f bacampidUin
hydrochloride CRS in 2 m L o f methanol R. Reference solution (a) Dissolve 30.0 m g o f bacampidllin
hydrochloride CRS in phosphate buffer A and dilute to
Reference solution (b) Dissolve 10 m g o f bacampidUin 100.0 m L w ith phosphate buffer A
hydrochloride CRS, 10 m g o f talampidUin hydrochloride CRS
and 10 mg o f pwampidUm CRS in 2 m l. o f methanol R.
Reference solution (b) D ilute 1.0 m L o f reference solution (a)
to 100.0 m L with phosphate buffer A.
Plate TLC silamsed silica gel plate R.
2016 Bacampicillin Hydrochloride 1-229
Reference solution (c) Dissolve 30 mg of the substance to be System suitability: reference solution (a):
examined in phosphate buffer B and dilute to 100 m L with — repeatability: maximum rdative standard deviation of
phosphate buffer B. H eat at 80 °C for about 30 min. 1.0 p er cent after 6 injections.
Reference solution (d) Dissolve 20 mg of ampidUin Calculate the percentage content of C 21H 28CIN 3O 7S from
trihydrate CRS (impurity I) in phosphate buffer A and dilute the d edared content of bacampicUHn hydrochloride CRS.
to 250 m L with phosphate buffer A. Dilute 5 m L o f this STORAGE
solution to 100 m L with phosphate buffer A.
In an airtight container.
Column:
— size. I = 0.05 m , 0 = 3.9 mm; IM P U R IT IE S
— stationary phase. octadecylsHyl silica gel for chromatography R
(5 |im ). H
Mobile phase Mix 30 volumes o f acetonitrile R l and ,CH3
70 volumes of a 0.06 per cent m/m solution o f
tetrahexylammonium hydrogen sulfate R in phosphate buffer B. H H
Flow rate 1.0 m lA nin.
Detection Spectrophotometer at 220 ran. A. (25,5i?,6JR)-6-amino-3,3-dimethyl-7-oxo-4-thia-l-
azabicydo[3.2.0]heptane-2-caiboxylic a d d
Injection 20 |iL o f the test solution and reference
( 6-am inopenidllanic ad d ),
solutions (b), (c) and (d).
Run time 3.5 times the retention time of bacampidllin. H NHa
System suitability".
— the peak due to impurity I is separated from the peaks
due to the solvent in the chromatogram obtained with
reference solution (d);
— relative retention with reference to bacampidllin: B. (2i?)-2-amino-2-phenylacetic a d d (D-phenylglycine),
degradation product eluting just after
bacam pidllin = 1.12 to 1.38 in the chromatogram
obtained with reference solution (c); if necessary, adjust
the concentration of tetrahexyiammonium hydrogen
sulfate in the mobile phase.
Limits:
— any impurity", for each impurity, not more than 1.5 times
the area of the principal peak in the chromatogram
obtained with reference solution (b) (1.5 p er cent); C . (2RS,45)-2-[[[(2i?)-2-annno-2-
— total: not more than 3 times the area o f the principal peak phenylacetyl] amino] methyl] -5,5-dimethylthiazolidine-4-
in the chromatogram obtained with reference solution (b) carboxylic a d d (penilloic adds of ampicillin),
(3 per cent);
— disregard limit. 0.1 times the area of the principal peak in *1 .c o 2h
the chromatogram obtained with reference solution (b) H NH2 H H N^C^CH a
(0.1 per cent).
B u ty l a c e ta te a n d eth y l a c e ta te (2.4.24> System A)
M axim um 2.0 per cent of butyl acetate, maximum
4.0 per cent of ethyl acetate and maximum 5.0 per cent for
the sum o f the contents. D . (45)-2-[[[(2JR)-2-amino-2-
Sample solution Dissolve 50.0 m g o f the substance to be phenylacetyl] amino] carboxymethyl]-535-dimethylthiazolidine-
examined in water R and dilute to 10.0 m L with the same 4-carboxylic a d d (penidUoic adds o f ampicillin),
solvent.
Use the method o f standard additions.
Static head-space conditions that may be used:
— equilibration temperature. 60 °C;
— equilibration time. 20 min.
iVyV-Dim etfaylaniline (2.4.26j, Method A)
M axim um 20 ppm .
W a te r (2.5.12)
M axim um 0.8 per cent, determined on 0.300 g. E. (4S)-2-(3,6-dioxo-5-phenylpiperazin-2-yl)-5,5-
S u lfa te d ash (2.4.14) dimethylthiazolidine-4-carboxylic a d d (diketopiperazines of
M axim um 1.5 p er cent, determined on 1.0 g. ampicillin),
A SSA Y
HS CH3
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modifications. H3C ^ X C° 2H anc*enantiomer
H NH2
Injection Test solution and reference solution (a).
F. (2ÄS)-2-amino-3-methyl-3-sulfanylbutanoic a d d
(DL-penicillamine),
1-230 Bacitracin 2016
H NH2
CHARACTERS
A p p e a ra n c e
W hite or almost white powder, hygroscopic.
S o lu b ility
G . methyl (2R)-2-amino-2-phenylacetate (methyl Freely soluble in w ater and in ethanol (96 p e r cent).
D -phenylgiycinate), ID E N T IF IC A T IO N
First identification B, C
H ? H- / H3i Second identification A , C
0 ^ 0 O' 'CH, A. Thin-layer chromatography (2.2.27).
Test solution Dissolve 10 m g o f the substance to be exam in ed
in a 3.4 g/L solution of hydrochloric add R and dilute to
and epimer at C* 1.0 m L with the sam e solution.
Reference solution Dissolve 10 m g o f bacitradn zinc CRS in a
H . ( IRS ) - 1- [(ethoxycarbonyl)oxy]ethyl (2.S,5/?,6i?)-6-[[(2.R)- 3.4 g/L solution o f hydrochloric acid R and dilute to 1.0 m L
2-(acetylamino)-2-phenylacetyl]amino]-3,3-dimethyl-7-oxo-4- w ith the same solution.
thia-l-azabicyclo[3.2.0]heptane-2-caiboxylate Plate TLC silica gd plate R.
(N-acetyibacampicillin), Mobile phase glacial acetic add R, water R, butanol R
(1:2:4 VIVIV).
o Application 10 |iL.
Development Over h alf o f the plate.
Drying A t 100-105 °C.
H H
Detection Spray with nmhydrm solution R1 and heat at 110 °C
for 5 min.
I. (2S,5i?,6i?)-6-[[(2i?)-2-amino-2-phenylacetyl]amino]-3,3-
Results T h e spots in the chrom atogram obtained with the test
solution are similar in position, size and colour to the spots
dim ethyl-7-oxo-4-thia-l-azabicydo [3.2.0]heptane-2-
in the chrom atogram obtained w ith the reference solution.
carboxylic a d d (am pidllin).
B. Composition (see Tests).
PhEur
C . Ignite 0.2 g. A n insignificant residue rem ains which is not
yellow at high tem perature. Allow to cool. Dissolve the
residue in 0.1 m L o f dilute hydrochloric add R. A dd 5 m L of
**★ water R and 0.2 m L of strong sodium hydroxide solution R.
Bacitracin * ★ N o white p redpitate is formed.
★ ★
(Ph. Eur. monograph 0465) * * * * * TESTS
S o lu tio n S
Dissolve 0.25 g in carbon dioxide-free water R and dilute to
h,n V H’ 25 m L with the sam e solvent.
A p p e a ra n c e o f so lu tio n
:' XN Solution S is clear (2.2.1).
p H (2.2.3)
L-Leu d-GIu Y—L- Lyso-OmX D-Phe-] 6.0 to 7.0 for solution S.
O t*- L-Asn— D-Asp -*• l-H is J C o m p o sitio n
L iquid chromatography (2.2.29): use the norm alisation
Name Mol. Formula X Y R procedure. Prepare the solutions immediately before use.
Bacitradn A CeeHi03Ni7OieS L-lle L-lle ch3 Test solution Dissolve 0.100 g of the substance to be
Bacitradn B1 CbsH io iN i 7 0 16S L-lle L-lle H e xam in ed in 50.0 m l . of the mobile phase.
Bacitradn B2 C «H io iN i70ieS L-Val L-lle ch3
Reference solution (a) Suspend 20.0 m g o f bacitradn zinc CRS
Bacitradn B3 C85H10iNi7O i6S L-lle L-Val ch3
in water R, add 0.2 m L o f dilute hydrochloric add R and dilute
to 10.0 m L with water R.
Column: R e la te d p e p tid e s
— size: I = 0.25 m , 0 = 4.6 mm; Liquid chromatography (2.2.29) as described in the test for
— stationary phase: end-capped octadecylsUyl silica gel for composition.
chromatography R (5 (im). See Figure 0465.-1.
Mobile phase A dd 40 volumes o f acetordtrile R, 300 volumes of Limit.
water R and 520 volumes of methanol R1 to 100 volumes o f a — sum of the areas of all peaks eluting before the peak due to
34.8 g/L solution o f dipotassium hydrogen phosphate R adjusted bacitracin Bl: maximum 20.0 per cent.
to p H 6.0 with a 27.2 g/L solution of potassium dihydrogen
Im p u rity E
phosphate R. Liquid chromatography (2.2.29) as described in the test for
Flow rate 1.0 m I7m in. composition.
Detection Spectrophotom eter at 254 nm. See Figure 0465.-2.
Injection 100 pL; inject the blank, the test solution and Detection Spectrophotometer at 254 nm; spectrophotometer
reference solutions (a) and (c). at 300 nm for reference solution (d).
Run time 3 times the retention time o f bacitracin A. Injection T est solution and reference solutions (b) and (d).
Relative retention W ith reference to bacitracin A (retention Limit:
time = 1 5 min to 25 min): bacitracin B1 = about 0.6; — impurity E: n o t more than 1.2 times the area o f the
bacitracin B3 = about 0.8; impurity E = about 2.5. principal peak in the chromatogram obtained with
If necessary, adjust the composition o f the mobile phase by reference solution (b) (6.0 per cent).
changing the am ount o f organic modifier whilst keeping the L oss o n d ry in g (2.2.32)
ratio constant betw een methanol and acetonitrile. M axim um 5.0 per cent, determined on 1.000 g by drying at
System suitability: reference solution (a): 60 °C over diphosphorus pentoodde R at a pressure not
— peak-to-valley ratio: minimum o f 1.2, where Hp = height exceeding 0.1 kPa for 3 h.
above the baseline o f the peak due to bacitracin B1 and S u lfa te d a s h (2.4.14)
Hv = height above the baseline o f the lowest point o f the M aximum 1.0 per cent, determined on 1.0 g.
curve separating this peak from the peak due to
bacitracin B2. S te rility
(2.6.1). If intended for the preparation of ophthalmic dosage
Limits:
forms without a further appropriate sterilisation procedure, it
— bacitracin A: minimnm 40.0 per cent;
complies with the test for sterility.
— sum of bacitracins A , B l, B2 and B3: minimum
70.0 per cent; B a c te ria l en d o to x in s (2.6.14)
— disregard limit: the area o f the peak due to bacitracin A in Less than 0.8 IU/mg, if intended for use in the manufacture
the chrom atogram obtained with reference solution (c) o f ophthalmic dosage forms without a further appropriate
(0.5 per cent); disregard any peak observed in the blank procedure for the removal of bacterial endotoxins.
run.
1-232 Bacitracin Zinc 2016
ASSAY ★ ★
Bacitracin Zinc ★ ★
Carry out the microbiological assay o f antibiotics (2.7.2).
*****
Use bacitracin zinc CRS as the reference substance. (Ph. Eur. monograph 0466)
STORAGE
Action and use
In an airtight container at 2 °C to 8 °C. If the substance is
Polypeptide antibacterial.
sterile, store in a sterile, airtight, tam per-proof container.
Preparations
IMPURITIES Polymyxin and Bacitracin O intm ent
Polymyxin and Bacitracin Eye O intm ent
PhEur.
DEFINITION
Zinc complex o f bacitracin, which consists o f a mixture o f
antimicrobial polypeptides produced by certain strains of
Bacillus lichemformis or Bacillus subtiHs, the m ain com ponents
A. X = L-Val, Y = L-Ile, R = H : bacitracin C l, being bacitracins A, B l, B2 and B3.
B. X = L -Ü e, Y = L-Val, R = H: bacitracin C2, Content
Minimum 60 IU /m g (dried substance).
C. X = Y = L-Val, R = C H 3: bacitracin C3,
D . X = Y = L-Val, R = H : bacitracin E, CHARACTERS
Appearance
W hite or light yellowish-grey powder, hygroscopic.
Solubility
Slightly soluble in water and in ethanol (96 p er cent).
IDENTIFICATION
First identification B, C
Second identification A , C
E. X = Y = L-Üe, R = C H 3: bacitracin F , A. Thin-layer chrom atography (2.2.27).
F. X = Y = L-Üe, R = H : bacitracin H I, Test solution Dissolve 10 m g o f the substance to be examined
in 0.5 m l. o f dilute hydrochloric add R and dilute to 1.0 m L
G . X = L-Val, Y = L-Üe, R = C H 3: bacitracin H 2,
with water R.
H . X = L-Ile, Y = L-Val, R = C H 3: bacitracin H 3,
Reference solution Dissolve 10 m g o f badtradn zinc CRS in
I. X = L-Val, Y = L-Üe, R = H : badtradn II, 0.5 m L o f dilute hydrochloric add R and dilute to 1.0 m L with
J. X = L-Ile, Y = L-Val, R = H : bacitracin 12, water R
K. X = Y = L-Val, R = C H 3: bacitracin 13. Plate TLC silica gel plate R.
PhEw
2016 Bacitracin Zinc 1-233
Mobile phase glacial acetic acid R, water R, butanol R Reference solution (d) Dissolve 50.0 mg of the substance to be
(1:2:4 VIVIV). examined in 25.0 m L of a 40 g/L solution of sodium edetate R
Application 10 |iL. adjusted to p H 7.0 with dilute sodium hydroxide solution R.
H eat in a boiling water-bath for 30 min. Cool to room
Development Over half of the plate.
temperature.
Drying h i 100-105 °C.
Blank solution A 40 g/L solution o f sodium edetate R adjusted
Detection Spray with ninhydrin solution R l and heat at 110 °C to p H 7.0 with dilute sodium hydroxide R.
for 5 min.
Column:
Residts T h e spots in the chromatogram obtained with the test — size: I = 0.25 m , 0 = 4.6 mm ;
solution are similar in position, size and colour to the spots — stationary phase: end-capped octadecylsHyl silica gel for
in the chrom atogram obtained with the reference solution. chromatography R (5 Jim).
B. Com position (see Tests). Mobile phase Add 520 volumes of methanol R l, 40 volumes of
C. Ignite about 0.15 g, allow to cool and dissolve the residue acetonitrile R and 300 volumes o f water R to 100 volumes of a
in 1 m L o f dilute hydrochloric acid R. Add 4 m L of water R. 34.8 g/L solution o f dipotassium hydrogen phosphate R,
T he solution gives the reaction of zinc (2.3.1). adjusted to p H 6.0 with a 27.2 g/L solution of potassium
TESTS dihydrogen phosphate R.
p H (2.2.3) Flow rate 1.0 mL/min.
6.0 to 7.5. Detection Spectrophotom eter at 254 nm.
Shake 1.0 g for about 1 min with 10 m L o f carbon dioxide-free Injection 100 nL; inject the blank, the test solution and
water R and filter. reference solutions (a) and (c).
C o m p o sitio n Run time 3 times the retention time of bacitracin A.
Liquid chromatography (2.2.29): use the normalisation Relative retention W ith reference to bacitracin A (retention
procedure. Prepare the solutions immediately before use. time = 15 min to 25 min): bacitracin B1 = about 0.6;
Test solution Dissolve 0.100 g of the substance to be bacitracin B3 = about 0.8; impurity E = about 2.5.
examined in 50.0 m L o f a 40 g/L solution of sodium edetate R If necessary, adjust the composition of the mobile phase by
adjusted to p H 7.0 with dilute sodium hydroxide solution R. changing the am ount o f organic modifier whilst keeping the
Reference solution (a) Dissolve 20.0 m g of bacitracin zinc CRS ratio constant between methanol and acetonitrile.
in 10.0 m L of a 40 g/L solution of sodium edetate R adjusted System suitability: reference solution (a):
to p H 7.0 with dilute sodium hydroxide solution R. — peak-to-valley ratio: minimum o f 1.2, where Hp = height
Reference solution (b) Dilute 5.0 m L of the test solution to above the baseline o f the peak due to bacitracin B1 and
100.0 m L with water R. Hv = height above the baseline o f the lowest point o f the
Reference solution (c) Dilute 1.0 m L o f reference solution (b) curve separating this peak from the peak due to
to 10.0 m L with water R. bacitracin B2.
1-234 Bacitracin Zinc 2016
Action and use Results T h e prindpal spot in the chromatogram obtained with
the test solution is similar in position, colour and size to the
Skeletal m u sd e relaxant
prindpal spot in the chromatogram obtained with the
Preparations reference solution.
Baclofen Oral Solution
TESTS
Baclofen Tablets
A p p e a ra n c e o f so lu tio n
PhEtr__________________________________________________________ T h e solution is n o t m ore opalescent than reference
suspension II (2.2.1) and not m ore intensely coloured than
D E F IN IT IO N
reference solution BY5 (2.2.2, Method II).
(3&S)-4-Amino-3-(4-chlorophenyl)butanoic ad d .
Dissolve 0.50 g in 1 M sodium hydroxide and dilute to 25 m L
Content with the same solvent.
98.0 p er cent to 101.0 per cent (anhydrous substance).
R e la te d su b sta n c e s
CHARACTERS Liquid chromatography (2.2.29).
Appearance Test solution Dissolve 25.0 mg o f the substance to be
W hite or almost white powder. examined in the mobile phase and dilute to 10.0 m L with
S o lu b ility the mobile phase.
Slightly soluble in water, very slightly soluble in ethanol
(96 per cent), practically insoluble in acetone. It dissolves in
1-236 Bambuterol Hydrochloride 2016
IM P U R IT IE S
Preparation Discs.
Specified impurities A Comparison bambuterol hydrochloride CRS.
Other detectable impurities (the following substances would, if If the spectra obtained show differences, dissolve the
present at a sufficient level, be detected by one or other of substance to be examined and the reference substance
the tests in the m onograph. T hey are limited by the general separatdy in a m ixture o f 1 volume o f water R and 6 volumes
acceptance criterion for other/unspecified impurities and/or o f acetone R, cool in ice to p redpitate and dry both
by th e general m onograph Substances for pharmaceutical use predpitates in vacuo at 50 °C to constant w tig h t Record new
(2034). It is therefore n o t necessary to identify these spectra using the residues.
impurities for dem onstration o f compliance. See also 5.10. B. It gives reaction (a) o f chlorides (2.3.1).
Control of impurities in substances for pharmaceutical use): B. TESTS
S o lu tio n S
Dissolve 4.0 g in carbon dioxide-free water R and dilute to
20.0 m L with the same solvent.
and enantiomer
A cid ity o r a lk a lin ity
H
T o 10 m L o f solution S add 0.2 m L o f methyl red sdution R
a and 0.2 m L o f 0.01 M hydrochloric acid. T h e solution is red.
A dd 0.4 m L o f 0.01 M sodium hydroxide. T h e solution is
A. (4RS)-4-(4-chlorophenyl)pyrrolidin-2-one, yellow.
O p tic a l r o ta tio n (2.2.7)
-0 .1 0 ° t o + 0.10°.
2016 Barbital 1-237
H OH
Dilute 1 m L o f solution S to 10 m L with carbon dioxide-free
water R.
and enantiomer
R e la te d su b sta n c e s
Liquid chromatography (2.2.29).
Test solution Dissolve 5.0 mg of the substance to be examined
in the mobile phase and dilute to 10.0 m L with the mobile
phase. A. R l = N H -C (C H 3)3, R2 = R3 = H: (li?S )-l-(3,5-
dihydroxyphenyl)-2-[(l,l-dimethylethyl)amino]ethanol
Reference solution (a) Dissolve 1.0 mg of formoterol jumarate
(terbutaline),
dihydrate CRS in die mobile phase and dilute to 10.0 m L
with the mobile phase. Mix 0.8 m L o f this solution with B. R l = O H , R2 = R3 = C O -N (C H 3)2: 5 -[(li?S )-l,2-
0.4 m L of the test solution and dilute to 100.0 m L with the dihydroxyethyl] -1,3-phenylene bis (dimethylcarb amate),
mobile phase. C . R l = N H -C (C H 3)3, R2 = H , R3 = C O -N (C H 3)2:
Reference solution (b) Dilute 1.0 m L of the test solution to 3- [(lftS )-2 -[(1,1 -dimethylethyl) amino] -1 -hydroxyethyl] -5-
50.0 m L with the mobile phase. Dilute 2.0 m L of this hydroxyphenyl dimethylcarbamate,
solution to 20.0 m L with the mobile phase. D . R l = H , R2 = R3 = C O -N (C H 3)2: 5-[(lRS)-l-
Column: hydroxyethyl]-l,3-phenylene bis(dimethylcarbamate),
— size: I = 0.15 m , 0 = 4.6 mm;
— stationary phase, base-deactivated octadecylsHyl silica gelfor ch3 o
chromatography R (5 |im).
Mobile phase Dissolve 1.3 g of sodium octanesulfonate R in
430 m L of a mixture of 25 volumes of acetonitrile R l and
75 volumes o f methanol R; then mix this solution with O
H3C" Y
570 m L of 0.050 M phosphate buffer p H 3.0 prepared as
follows: dissolve 6.90 g of sodium dütydrogen phosphate
monohydrate R in water R and dilute to 1000 m L with
E. R = H : 5-acetyl-l,3-phenylene bis(dimethylcarbamate),
water R, adjust to p H 3.0 with a 50 g/L solution of dilute
phosphoric acid R. F . R = N H -C (C H 3)3: 5- [ [( 1,1 -dimethylethyl)amino] acetyl] -
1,3-phenylene bis (dimethylcarbamate).
Flow rate 1.5 mL/min.
PhEur
Detection Spectrophotometer at 214 nm.
Injection 20 |iL; inject the mobile phase as a blank.
Run time 1.5 times the retention time o f bambuterol.
Retention time Form oterol = about 7 min; ★ ★
bambuterol = about 9 min. If necessary, adjust the Barbital ★ ★
composition o f the mobile phase; increase the content o f *****
(Ph. Ever, monograph 0170)
phosphate buffer to increase the retention time.
System suitability: reference solution (a):
— resolution: m in im u m 5.0 between the peaks due to
bambuterol and formoterol.
Limits:
— impurities A , B, C, D, E, F: for each impurity, n o t more
than the area o f the principal peak in the chromatogram
obtained w ith reference solution (b) (0.2 per cent); C8Hi2N 20 3 184.2 57-44-3
— total: not m ore than 3 times the area o f the principal peak
in the chromatogram obtained with reference solution (b) A c tio n a n d u se
(0.6 per cent); Barbiturate.
— disregard limit. 0.25 times the area of the principal peak in
PhEur____________
the chromatogram obtained with reference solution (b)
(0.05 per cent); disregard any peak due to the mobile D E F IN IT IO N
phase. Barbital contains not less than 99.0 per cent and not m ore
W a te r (2.5.12) than the equivalent of 101.0 per cent of
M aximum 0.5 p er cent, determined on 0.500 g. 5,5-diethyipyrimidine-2,4,6(lH,3H,5.H)-trione, calculated
with reference to die dried substance.
S u lfa te d a sh (2.4.14)
M aximum 0.1 p er cent, determined on 1.0 g. CHARACTERS
A white or alm ost white, crystalline powder or colourless
A SSAY
crystals, slightly soluble in water, soluble in boiling water and
Dissolve 0.320 g in 50 m L o f ethand (96 per cent) R and add
in alcohol. It forms water-soluble compounds with alkali
5 m L of 0.01 M hydrochloric acid. C an y out a potentiometric
hydroxides and carbonates and w ith ammonia.
titration (2.2.20), using 0.1 M sodium hydroxide. Read
the volume added between the 2 points o f inflexion. ID E N T IF IC A T IO N
1 m L of 0.1 M sodium hydroxide is equivalent to 40.39 mg of First identification A , B
C 13H 30CIN 3O 5. Second identification A, C, D.
IM P U R IT IE S A. Determ ine the melting point (2.2.14) of the substance to
be examined. M ix equal parts o f the substance to be
Specified impurities A, B, C, D , E, F.
examined and barbital CRS and determine the melting point
1-238 Barium Sulfate 2016
o f the mixture. T he difference between the melting points in pyridine R. T itrate with 0.1 M ethanolic sodium hydroxide
(which are about 190 °C) is n o t greater than 2 °C. until a pure blue colour is obtained. Carry o u t a blank
B. Examine by infrared absorption spectrophotometry titration.
(2.2.24), comparing with the spectrum obtained with 1 m L o f 0.1 M ethanolic sodium hydroxide is equivalent to
barbital CRS. 9.21 m g o f C8H 12N 20 3.
C. Examine by thin-layer chrom atography (2.2.27), using PhEtr
silica gel GF254 R as the coating substance.
Test solution Dissolve 75 m g o f the substance to be examined
in alcohol R and dilute to 25 m L with the same solvent.
Reference solution Dissolve 75 m g o f barbital CRS in alcohol R Barium Sulfate ★ ★
★ ★
and dilute to 25 m L with the same solvent.
(Ph Eiar monograph 0010) *****
Apply separately to the plate 10 |iL of each solution. Develop
over a path o f 18 cm using the lower layer o f a mixture o f BaS0 4 233.4 7727-43-7
5 volumes of concentrated ammonia R, 15 volumes o f alcohol R A c tio n a n d u se
and 80 volumes o f chloroform R. Examine immediately in Radio-opaque substance used in the investigation o f the
ultraviolet light at 254 nm. T h e principal spot in the gastro-intestinal tra c t
chrom atogram obtained with the test solution is similar in
position and size to the principal spot in the chromatogram P re p a r a tio n
obtained with the reference solution. Barium Sulfate for Suspension
A c tio n a n d use
Anhydrous Beclometasone *****
Radio-opaque preparation used in the investigation of the Dipropionate *****
gastro-intestinal tract. (Ph. Eur. monograph 0654)
P re p a r a tio n
Barium Sulfate Oral Suspension O
D E F IN IT IO N
Barium Sulfate for Suspension is a dry mixture of Barium
Sulfate with a suitable dispersing agent and may contain
suitable flavours and suitable antimicrobial preservatives.
C o n te n t o f b a riu m sulfate, B a S 0 4
90.0 to 110.0% o f the stated amount.
C H A R A C T E R IS T IC S
A fine, white or creamy white powder.
ID E N T IF IC A T IO N A ctio n a n d u se
A Ignite 1 g to constant weight. T o 0.2 g of the residue add Glucocorticoid.
5 m L o f a 50% w/v solution o f sodium carbonate and boil for P re p a ra tio n s
5 m in u tes. Add 10 m l. of water and filter. Reserve the
Beclometasone Cream
residue for test B. Acidify a portion of the filtrate with
Beclometasone Aqueous Nasal Spray
2m hydrochloric acid. T he solution yields the reactions
characteristic of sulfates, Appendix VI. Beclometasone Inhalation Powder
B. W ash the residue reserved in test A with water, add 5 m L Beclometasone Inhalation Powder, pre-dispensed
of 2m hydrochloric acid, m ix well and filter. Add 0.3 m L o f Beclometasone O intm ent
1m sulfuric acid to the filtrate. A white precipitate is produced Beclometasone Pressurised Inhalation
which is insoluble in 2m hydrochloric acid.
PhEur______________________________________________________
TESTS
A c id ity o r alkalinity D E F IN IT IO N
p H o f an aqueous suspension containing the equivalent o f 9-Chloro-l 1ß-hydroxy-16ß-methyi-3,20-dioxopregna-l,4-
60% w/w o f Barium Sulfate or, for lower strengths, the diene-17,21-diyl dipropanoate.
aqueous suspension at the strength o f intended use, 3.5 to C o n te n t
8.5, Appendix V L. 96.0 per cent to 102.0 per cent (dried substance).
L oss o n d ry in g CHA RACTERS
W hen dried at 105° for 4 hours, loses not more than 1.0% of A p p e a ra n c e
its weight. Use 1 g. White or almost white, crystalline powder.
1-240 Beclometasone Dipropionate 2016
J. R1 = R2 = CO -C 2H 5: 9,lip-epoxy-16p-m ethyl-3,20-
dioxo-9 P-pregna-1,4-diene-17,21 -diyl dipropanoate,
R. R1 = R2 = H : 9,11 p-epoxy-17,2 1-dihydroxy-l 6 P-methyl-
9P-pregna-l,4-diene-3,20-dione,
U . R1 = C O -C 2H 5, R2 = H: 9,HP-epoxy-21-hydroxy-16P-
methyl-3,20-dioxo-9p-pregna-l,4-dien-17-yl propanoate,
A. R1 = R3 = H , R2 = Cl, R4 = C O -Q H g: 9-chloro-
V. R 1 = H , R2 = CO-C 2H 5: 9,llp-epoxy-17-hydroxy-16P-
lip,17-dihydroxy-16p-m ethyl-3,20-dioxopregna-l,4-dien-21-
methyl-3,20-dioxo-9P-pregna-l,4-dien-2 1-yl propanoate,
yl propanoate (beclometasone 21-propionate),
B. R1 = H , R2 = Cl, R3 = CO-C2H 5, R4 = C O -C H 3: o
2 l-(acetyloxy)-9-chloro-l 1P-hydroxy-16P-methyi-3,20-
dioxopregna-1,4-dien-17-yl propanoate (beclometasone
21-acetate 17-propionate),
C. R1 = H , R2 = Cl, R3 = CO-C2H 5, R4 = C O -C H 2-
C H 2-C H 3: 9-chloro-l 1P-hydroxy-16P-methyl-3,20-dioxo-l 7-
(propanoyloxy)-pregna-1,4-dien-21-yl butanoate
(beclometasone 21-butyrate 17-propionate),
D . R1 = H , R2 = Br, R3 = R4 = C O -Q H g: 9 -b ro m o -llp - L. 9-chloro-l 1P-hydroxy-16P-methyl-3,20-dioxopregn-4-ene-
hydroxy-16 P-methyl-3^0-dioxopregna-1,4-diene-17,21 -diyl 17,21-diyl dipropanoate,
dipropanoate,
o
F . R1 = Br, R2 = Cl, R3 = R4 = C O -Q H g: 6a-bromo-9-
chloro-11P-hydroxy-16P-methyl-3,20-dioxopregna-l,4-diene-
17,21-diyl dipropanoate,
M . 9-chloro-l 1P-hydroxy-16P-methyl-3,20-dioxopregna-4,6-
diene-17,21-diyl dipropanoate,
o
E. R1 = Cl, R2 = CO -C 2H 5: 6a,9-dichloro-lip-hydroxy-
16p-methyl-3,20-dioxopregna-l ,4-diene-l 7,21 -diyl
dipropanoate,
H . R1 = R2 = H : 9-chloro-l ip ,2 1-dihydroxy-l 6 P-methyl-
3,20-dioxopregna-l,4-dien-17-yl propanoate (beclometasone
17-propionate),
N . 2-bromo-9-chloro-l 1P-hydroxy-16P-methyl-3,20-
dioxopregna-1,4-diene-17,21-diyl dipropanoate,
O
O
0 ^ y ° 0 'w-CH,
ChT H T \-C H 3
I. 16P-methyi-3,20-diQxopregna-l,4,9(l l)-triene-17,21-diyl
O. R1 = R2 = Cl: 9,liP-dichloro-16P-methyl-3,20-
dipropanoate,
dioxopregna-l,4-diene-17,21-diyl dipropanoate,
Q. R1 = R2 = H : 16P-methyl-3,20-dioxopregna-l,4-diene-
17,21-diyl dipropanoate,
S. R1 = O -C O -C 2H 5, R2 = Cl: 9-chloro-l6p-m ethyl-3,20-
dioxopregna-l,4-diene-l ip,17,21-triyl tripropanoate
(beclometasone tripropionate).
__________________________________________________________ PhEur
1-242 Beclometasone Dipropionate Monohydrate 2016
D E F IN IT IO N 4 -1 2 40-» 45 60 55
9-C hloro-l 1{5-hydroxy-16 P-methyi-3,20-dioxopregna-1,4- 12 -5 9 45 55
diene-17,21-diyl dipropanoate monohydrate.
Content Flow rate 1.4 mL/min.
97.0 per cent to 102.0 p er cent (dried substance). Detection Spectrophotom eter at 254 nm .
CHARACTERS Injection 20 pi o f test solution (a) and reference solutions (a),
Appearance (b) and (c).
W hite or alm ost w hite powder. Identification of impurities U se the chrom atogram supplied
Solubility with beclometasone dipropionate for peak identification CRS and
Practically insoluble in water, freely soluble in acetone, the chrom atogram obtained with reference solution (c) to
sparingly soluble in ethanol (96 per cent). identify the peaks due to impurities B, C , F and L; use the
chrom atogram supplied with beclometasone dipropionate for
ID E N T IF IC A T IO N system suitability CRS and the chrom atogram obtained with
A. Infrared absorption spectrophotom etry (2.2.24). reference solution (b) to identify the peak due to impurity D .
Comparison beclometasone dipropionate monohydrate CRS. Relative retention W ith reference to beclometasone
B. T reat 25 m g by th e oxygen-flask m ethod (2.5.10). U se a dipropionate (retention time = about 25 min):
mixture of 1 m L o f 1 M sodium hydroxide and 20 m L o f im purity B = about 0.6; im purity D = about 1.1;
water R to absorb the com bustion products. T h e solution impurity L = about 1.3; im purity C = about 1.8;
gives reaction (a) o f chlorides (2.3.1). im purity F = about 2.2.
C. Loss on drying (see Tests). System suitability: reference solution (b):
— peak-to-vaHey ratio: m inim um 1.5, where Hp = height
TESTS
above the baseline o f the peak due to im purity D and
Specific optical rotation (2.2.7) Hv — height above the baseline o f the lowest point o f the
+ 108 to + 115 (dried substance).
curve separating this peak from the peak due to
Dissolve 0.100 g in ethanol (96 per cent) R and dilute to beclom etasone dipropionate.
10.0 m L with the sam e solvent. Limits:
Related substances — correction factor, for the calculation o f content, multiply the
Liquid chrom atography (2.2.29). peak area o f im purity F by 1.3;
Solvent mixture M obile phase A, mobile phase B (45:55 VIV). — impurity B: n o t m ore than 5 times the area o f the
Test solution (a) Dissolve 50.0 m g o f the substance to be principal peak in the chrom atogram obtained with
examined in 28 m L o f mobile phase B and dilute to 50.0 m L reference solution (a) (0.5 per cent);
with mobile phase A. — impurities C, F, L: for each impurity, not m ore than
1 .5 times the area o f the principal peak in the
Test solution (b) D ilute 1.0 m L o f test solution (a) to
chrom atogram obtained w ith reference solution (a)
50.0 m L with the solvent mixture.
(0.15 p er cent);
Reference solution (a) D ilute 5.0 m L o f test solution (b) to — unspecified impurities: for each impurity, n o t more than the
100.0 m L with the solvent mixture. area of the principal peak in the chrom atogram obtained
Reference solution (b) Dissolve 5 m g o f beclometasone w ith reference solution (a) (0.10 per cent);
dipropionate for system suitability CRS (containing im purity D ) — total: n o t m ore than 10 times the area of the principal
in 3 m L o f mobile phase B and dilute to 5 m L with mobile peak in die chrom atogram obtained with reference
phase A. solution (a) (1.0 per cent);
2016 Beclometasone Dipropionate Monohydrate 1-243
on °joA .\ -
-R 2
M . 9-chloro-l 1P-hydroxy-16P-methyl-3,20-dioxopregna-4,6-
diene-17 ,2 1-diyl dipropanoate,
H R1
B. R I = H , R2 = C O C H 3: 21-(acetyloxy)-9-chloro-lip-
hydroxy-16 P-methyl-3,2 0-dioxopregna-1,4-dien-17-yl
propanoate (beclometasone 21-acetate 17-propionate),
C. R I = H , R2 = C O -C H 2-C H 2-C H 3: 9-chloro-l lp -
hydroxy-16P-methyl-3,20-dioxo-l 7-(propanoyloxy)-pregna-
l,4-dien-21-yl butanoate (beclometasone 21-butyrate N . R I = Br, R2 = O H , R3 = Cl: 2-brom o-9-chloro-l 1P-
17-propionate), hydroxy-16P-methyl-3,20-dioxopregna-l,4-diene-l 7,2 1-diyl
F. R I = Br, R2 = CO-C 2H 5: 6a-brom o-9-chloro-lip- dipropanoate,
hydroxy-16 P-methyl-3,20-dioxopregna- 1,4-diene-17,21 -diyl O. R I = H , R2 = R3 = Cl: 9,11 P-dichloro-16P-methyl-
dipropanoate, 3,20-dioxopregna-l,4-diene-17,21-diyl dipropanoate,
1-244 Beeswax 2016
Q. R l = R2 = R3 = H : 16P-methyl-3,20-dioxopregna-l,4- of ethanol (96 per cent) R and xylene R and a few glass beads.
diene-17,21-diyl dipropanoate, H eat until the substance is dissolved. Add 25.0 m L o f 0.5 M
S. R l = H , R2 = 0 - C 0 - C 2H 5, R3 = Cl: 9-chloro-16p- alcoholic potassium hydroxide and heat under a reflux
methyl-3,20-dioxopregna-l ,4 -d ien e-lip ,1 7 321-triyl condenser for 3 h. Titrate the hot solution immediately with
tripropanoate (bedom etasone tripropionate). 0.5 M hydrochloric acid, using 1 m L o f phenolphthalein
solution R l as indicator (nx mL). Reheat the solution to
__________________________________________________________ PhEur
boiling several times during the course o f the titration. Carry
out a blank test (n2 mL).
.* * * ★ 28.05 (na — m )
★ Saponification value =
White Beeswax ★ ★
★+ ★
(Ph. Eur. monograph 0069)
Ceresin, paraffins and certain other waxes
Action and use T o 3.0 g, in a 100 m L round-bottom ed flask, add 30 m L of
Excipient a 40 g/L solution o f potassium hydroxide R in aldehyde-free
alcohol R and boil gently under a reflux condenser for 2 h.
PhEur_______________________
Remove the condenser and immediately insert a
DEFINITION thermom eter. Place the flask in a water-bath at 80 °C and
Wax obtained by bleaching yellow beeswax. allow to cool, swirling the solution continuously.
N o precipitate is formed until 65 °C, although the solution
CHARACTERS
may be slightly opalescent. Beginning at 65 °C, the solution
Appearance may become cloudy and precipitates may be formed.
White or yellowish-white pieces or platesj translucent when
At 59 °C, the solution is cloudy.
thin, with a fine-grained, m att and non-crystalline fracture;
when w anned in the hand they become soft and malleable. Glycerol and other polyols
M axim um 0.5 per cent m/m, calculated as glycerol.
It has an odour similar to that o f yellow beeswax, though
fainter and never rancid. It is tasteless and does not stick to T o 0.20 g add 10 m L o f alcoholic potassium hydroxide
the teeth. solution R and heat on a water-bath under a reflux condenser
for 30 min. Add 50 m L o f dilute sulfuric add R, cool and
Solubility
filter. Rinse the flask and the filter with dilute sulfuric add R.
Practically insoluble in water, partially soluble in hot ethanol
Combine the filtrate and washings and dilute to 100.0 m L
(90 p er cent V/V) and completely soluble in fatty and
w ith dilute sulfuric add R. Place 1.0 m L of the solution in a
essential oils.
test-tube, add 0.5 m L o f a 10.7 g/L solution o f sodium
Relative density periodate R, mix and allow to stand for 5 min. Add 1.0 m L of
About 0.960. decolorisedfuchsin solution R and mix. Any precipitate
TESTS disappears. Place the tube in a beaker containing water at
Drop point (2.2.17) 40 °C. D uring cooling observe for 10-15 min. Any violet-
61 °C to 66 °C. blue colour in the solution is not more intense than that in a
standard prepared at the same time and in the same m anner
M elt the beeswax by heating on a water-bath, pour onto a
using 1.0 m L of a 10 m g/L solution o f glycerol R in dilute
glass plate and allow to cool to a semi-solid mass. Fill the
sulfuric add R.
metal cup by inserting the wider end into the beeswax and
__________________________________________________________ PhEur
repeating the procedure until beeswax extrudes from the
narrow opening. Remove the excess with a spatula and insert
the therm om eter immediately. Remove the beeswax
displaced. Allow to stand at room temperature for at least
12 h before determining the drop point. *****
Yellow Beeswax ★ ★
Acid value *****
17.0 to 24.0. (Ph. Eur. monograph 0070)
To 2.00 g (m g), in a 250 m L conical flask fitted with a
Action and use
reflux condenser, add 40 m L of xylene R and a few glass
Excipient.
beads. H eat until the substance is dissolved. Add 20 m L of
ethanol (96 per cent) R and 0.5 m L o f phenolphthalein PhEur.
solution R l and titrate the hot solution with 0.5 M alcoholic
DEFINITION
potassium hydroxide until a red colour persists for at least 10 s
Wax obtained by melting the walls o f the honeycomb made
(«i mL). Carry out a blank test («2 mL).
by the honey-bee, Apis meUifera L., with hot water and
removing foreign matter.
Add value = 28-°^ (ni ~ ni)
tn CHARACTERS
Appearance
Ester value (2.5.2) Yellow or light brown pieces or plates with a fine-grained,
70 to 80. m att and non-crystalline fracture; when w anned in the hand
they become soft and malleable.
Saponification value
87 to 104. It has a faint odour, characteristic of honey. It is tasteless and
does n o t stick to the teeth.
To 2.00 g (m g), in a 250 m L conical flask fitted with a
reflux condenser, add 30 m L o f a mixture o f equal volumes
2016 Benazepril Hydrochloride 1-245
S olubility filter. Rinse the flask and the filter with dilute sulfuric acid R.
Practically insoluble in water, partially soluble in h o t ethanol Combine the filtrate and washings and dilute to 100.0 m L
(90 per cent V/V) and completely soluble in fatty and with dilute sulfuric add R. Place 1.0 m l. o f the solution in a
essential oils. test-tube, add 0.5 m L of a 10.7 g/L solution of sodium
R elativ e d e n sity periodate R, mix and allow to stand for 5 min. Add 1.0 m L of
A bout 0.960. decolorisedfuchsm solution R and mix. Any precipitate
disappears. Place the tube in a beaker containing water at
TESTS 40 °C. D uring cooling observe for 10-15 min. Any violet-
D ro p p o in t (2.2.17) blue colour in the solution is not more intense than that in a
61 °C to 66 °C. standard prepared at the same time and in the same m anner
M elt the beeswax by heating on a water-bath, p our onto a using 1.0 m L of a 10 mg/L solution o f glycerol R in dilute
glass plate and allow to cool to a semi-solid mass. Fill the sulfuric add R.
metal cup by inserting the wider end into the beeswax and __________________________________________________________ PhEur
repeating the procedure until beeswax extrudes from the
narrow opening. Remove the excess with a spatula and insert
the therm om eter immediately. Remove the beeswax
displaced. Allow to stand at room tem perature for at least
12 h before determining the drop point. Benazepril Hydrochloride ****%
A cid value *. *
(Ph. Eur. monograph 2388) *
17.0 to 22.0.
T o 2.00 g (m g), in a 250 m L conical flask fitted with a
reflux condenser, add 40 m L of xylene R and a few glass
beads. H eat until the substance is dissolved. Add 20 m L o f
ethanol (96 per cent) R and 0.5 mL of phenolphxhalein
solution R1 and titrate the hot solution with 0.5 M alcoholic
potassium hydroxide until a red colour persists for at least 10 s
(ni m L). Carry out a blank test («2 mL).
C24H 29CIN 2O 5 461.0 86541-74-4
A #J , 28.05 ( n i - n 2)
A cid value = ---------------------
m A c tio n a n d u se
Angiotensin converting enzyme inhibitor.
E s te r v alu e (2.5.2)
PhEir__________________________________________________________
70 to 80.
S ap o n ific a tio n v alu e D E F IN IT IO N
87 to 102. [(3S)-3-[ [( 1S )-1-(Ethoxycarbonyl)-3-phenylpropyl] amino] - 2 -
T o 2.00 g (m g), in a 250 m L conical flask fitted with a oxo-2}3,4,5-tetrahydro- 1/f-l-benzazepin- 1-yl] acetic a d d
reflux condenser, add 30 m L of a mixture o f equal volumes hydrochloride.
o f ethanol (96 per cent) R and xylene R and a few glass beads. C o n te n t
H eat until the substance is dissolved. Add 25.0 m L o f 0.5 M 97.5 per cent to 102.0 per cent (dried substance).
alcoholic potassium hydroxide and heat under a reflux CHARACTERS
condenser for 3 h. T itrate the hot solution immediately with
A p p e a ra n c e
0.5 M hydrochloric acid, using 1 m L o f phenolphthalein W hite or almost white, crystalline powder, hygroscopic.
solution R1 as indicator («j mL). Reheat the solution to
boiling several times during the course o f the titration. Carry S o lu b ility
out a blank test («2 mL)- Slightly soluble in water, freely soluble in anhydrous ethanol,
very slightly soluble in ethyl acetate, practically insoluble in
cyclohexane.
S ap o n ificatio n value = It shows polymorphism (5.9).
m
ID E N T IF IC A T IO N
C e re sin , p a ra ffin s a n d c e rta in o th e r w axes Carry out either tests A, B, D or tests B, C, D.
T o 3.0 g, in a 100 m L round-bottom ed flask, add 30 m L of A. Specific optical rotation (2.2.7): —141 to -1 3 6 (dried
a 40 g/L solution of potassium hydroxide R in aldehyde-free substance).
alcohol R and boil gently under a reflux condenser for 2 h.
Dissolve 1.000 g in anhydrous ethanol R and dilute to
Remove the condenser and immediately insert a
50.0 m L with the same solvent.
therm om eter. Place the flask in a water-bath at 80 °C and
allow to cool, swirling the solution continuously. B. Infrared absorption spectrophotometry (2.2.24).
N o precipitate is formed until 65 °C, although the solution Comparison benazepril hydrochloride CRS.
may be slightly opalescent. Beginning at 65 °C, the solution If the spectra obtained in the solid state show differences,
may become cloudy and precipitates may be formed. dissolve the substance to be examined and the reference
At 59 °C, the solution is cloudy. substance separately in methanol R, evaporate to dryness and
G lycerol a n d o th e r polyols record new spectra using the residues.
M aximum 0.5 per cent m/m, calculated as glycerol. C. Enantiomeric purity (see Tests).
T o 0.20 g add 10 m L o f alcoholic potassium hydroxide D . It gives reaction (a) o f chlorides (2.3.1).
solution R and heat on a water-bath under a reflux condenser
for 30 min. A dd 50 m L o f dilute sulfuric acid R, cool and
1-246 Benazepril Hydrochloride 2016
TESTS — disregard limit. 0.25 times the area of the principal peak in
Related substances the chromatogram obtained with reference solution (c)
Liquid chromatography (2.2.29). (0.05 per cent).
Test solution (a) Dissolve 50.0 mg o f the substance to be E n a n tio m e ric p u rity
examined in the mobile phase and dilute to 50.0 m L with Liquid chromatography (2.2.29).
the mobile phase. Buffer solution pH 60 Dissolve 3.58 g o f disodium hydrogen
Test solution (b) Dilute 10.0 m L o f test solution (a) to phosphate R and 9.66 g o f potassium dihydrogen phosphate R in
100.0 m L with the mobile phase. water R and dilute to 1000.0 m L with the same solvent
Reference solution (a) Dissolve 50.0 mg o f benazepril Test solution Dissolve 50.0 mg o f the substance to be
hydrochloride CRS in the mobile phase and dilute to 50.0 m L examined in the mobile phase and dilute to 50.0 m L with
with the mobile phase. Dilute 10.0 m L of this solution to the mobile phase.
100.0 m L with the mobile phase. Reference solution (a) Dissolve 5.0 m g o f benazepril
Reference solution (b) Dissolve the contents of a vial of impurity A CRS in the mobile phase and dilute to 50.0 m L
benazeprilfor system suitability CRS (containing impurities B, with the mobile phase.
C, D , E, F and G) in 1.0 m L of test solution (a). Reference solution (b) Dilute 1.0 m L o f reference solution (a)
Reference solution (c) Dilute 1.0 m L of reference solution (a) to 100.0 m L with the mobile phase.
to 50.0 m L with the mobile phase. Reference solution (c) Dilute 1.0 m L of reference solution (a)
Column: to 10.0 m L with the mobile phase. Dilute 1.0 m L o f this
— size: I = 0.30 m, 0 = 3.9 mm ; solution to 10.0 m L with the test solution.
— stationary phase: end-capped octadecylsüyl silica gelfor Column:
chromatography R (10 pm). — size: I = 0.10 m , 0 = 4.0 mm;
Mobile phase Add 0.2 m L of glacial acetic acid R to 1000 m L — stationary phase: spherical silica gel AGP for chiral
o f a mixture o f 360 volumes o f water R and 640 volumes of chromatography R (5 pm);
methanol R2; add 0.81 g o f tetrabutylammonium bromide R and — temperature: 30 °C.
stir to dissolve. Mobile phase methanol R2, buffer solution p H 6.0
Flow rate 1.0 mlVmin. (20:80 V/V).
Detection Spectrophotometer at 240 nm . Flow rate 0.9 mL/min.
Injection 25 pL of test solution (a) and reference solutions (b) Detection Spectrophotom eter at 240 nm .
and (c). Injection 50 pL of the test solution and reference solutions (b)
Run time 3 times the retention time o f benazepril. and (c).
Relative retention W ith reference to benazepril (retention Run time 3.5 times the retention tim e of benazepril.
time = about 6 min): impurity E = about 0.3; Relative retention W ith reference to benazepril (retention
impurity F = about 0.4; impurity C = about 0.5; time — about 6 min): impurity A = about 1.9.
impurity B = about 1.8; impurity D = about 2.0; System suitability: reference solution (c):
impurity G = about 2.5. — peak-to-valley ratio: minimum 2.5, where Hp = height
Identification of impurities Use the chromatogram supplied above the baseline o f the peak due to impurity A and
with benazepril for system suitability CRS and the Hv = height above the baseline o f the lowest point o f the
chrom atogram obtained with reference solution (b) to curve separating this peak from the peak due to
identify the peaks due to impurities B, C, D , E, F and G. benazepril.
System suitability.: reference solution (b): Limit.
— resolution: m in im u m 2.5 between the peaks due to — impurity A: n o t more than the area of the corresponding
benazepril and impurity B and minimum 1.5 between the peak in the chromatogram obtained with reference
peaks due to impurities E and F. solution (b) (0.1 per cent).
Limits: H eav y m e ta ls (2.4.8)
— correction factors: for the calculation o f content, multiply M aximum 20 ppm.
the peak areas of the following impurities by the
1.0 g complies with test C. Prepare the reference solution
corresponding correction factor, impurity E = 0.5;
using 2 mT. o f lead standard solution (10 ppm Pb) R.
im purity F = 0.7;
— impurity B: n o tm o re than 2.5 times the area o f the L oss o n d ry in g (2.2.32)
principal peak in the chromatogram obtained with M aximum 1.5 per cent, determined on 1.000 g by drying
reference solution (c) (0.5 per cent); in vacuo at 105 °C for 3 h.
— impurity C: not m ore than 1.5 times die area of the S u lfa te d a s h (2.4.14)
principal peak in the chromatogram obtained with M aximum 0.1 per cent, determined on 1.0 g.
reference solution (c) (0.3 per cent);
A SSA Y
— impurities D, E, F, G: for each impurity, not more than
Liquid chromatography (2.2.29) as described in the test for
the area o f the principal peak in the chromatogram
related substances with the following modification.
obtained with reference solution (c) (0.2 per cent);
— unspecified impurities: for each impurity, not more than Irqection T est solution (b) and reference solution (a).
0.5 times the area o f the principal peak in the Calculate the percentage content o f C 24H 29CIN 2O 5 from the
chrom atogram obtained with reference solution (c) declared content o f benazepril hydrochloride CRS.
(0.10 per cent);
STORAGE
— total: not m ore than 10 times the area of the principal
Protected from light, in an airtight container.
peak in the chromatogram obtained with reference
solution (c) (2.0 per cent);
2016 Bendroflumethiazide 1-247
IM P U R IT IE S
Bendroflumethiazide ★
★
★
★
Specified impurities A, B, C, D , E 3 F 3 G
(Ph. Eitr. monograph 0370) *****
O O O 0
* '/ * //
C02h
H2n
,s s.
‘NH
and enantiomer
f3c A -
H H
PhEir.
D E F IN IT IO N
B. [(3i?S)-3-[[(1 SR)- 1-(ethoxycarbonyl)-3- (3^5)-3-Benzyl-6-(trifluorom ethyl)-3,4-dihydro-2H-l,2,4-
phenylpropyl]am ino]-2-oxo-2,3j4j5-tetrahydro-lii-l- benzothiadiazine-7-sulfonamide 1, 1-dioxide.
benzazepin-l-yl] acetic acid.
C o n te n t
98.0 p er cent to 102.0 per cent (dried substance).
o
H H || CHARACTERS
A p p e a ra n c e
W hite o r almost white, crystalline powder.
S o lu b ility
Practically insoluble in water, freely soluble in acetone,
soluble in ethanol (96 per cent).
C. R = H: (2S)-2-[[(3<S)-l-(carboxymethyl)-2-oxo-2,3,4,5-
tetrahydro- IH -l -benzazepin-3-yI] amino] -4-phenyIbutanoic ID E N T IF IC A T IO N
acid, Infrared absorption spectrophotometry (2.2.24).
G. R = C 2H 5: ethyl (25)-2-[[(35)-l-(2-ethoxy-2-oxoethyl)-2- Comparison bendroflumethiazide CRS.
oxo-2,3,4 j5-tetrahydro- IH- l-benzazepin-3-yl] amino] -4- TESTS
phenylbutanoate, R e la te d su b sta n c e s
Liquid chrom atography (2.2.29). Prepare the solutions
immediately before use.
Solvent mixture M ix 40 volumes of methanol R and
CO,H
60 volumes of a 2.0 gfL solution o f citric acid R.
Test solution Dissolve 10.0 mg of the substance to be
examined in the solvent mixture and dilute to 50.0 m L w ith
the solvent mixture.
D . [(35)-3-[[(15)-3-cyclohexyl-1- Reference solution (a) Dissolve 2 m g of bendroflumethiazide
(ethoxycarbonyl)propyl]amino]-2-oxo-2,3,4,5-tetrahydro-lfi- impurity A CRS and 2.5 mg of altizide CRS in the solvent
1-benzazepin-1-yl] acetic ad d , m ixture and dilute to 10 m L with the solvent mixture.
M ix 1 m L of this solution with 1 m L of the test solution and
dilute to 100 m L with the solvent mixture.
H,N. Reference solution (b) Dilute 1.0 m l. of the test solution to
100.0 m L with the solvent mixture. Dilute 1.0 m L of this
solution to 10.0 m L with the solvent mixture.
Column:
— size: I = 0.15 m , 0 = 3.0 mm;
— stationary phase: end-capped octadecylsUyl silica gel for
E. R = H: [(35)-3-am ino-2-oxo-2^,4,5-tetrahydro-lH -l-
chromatography R (5 jam);
benzazepin-l-yl] acetic add,
— temperature: 40 °C.
F. R = C (C H 3) 3: 1,1-dimethylethyl [(3S)-3-amino-2-oxo-
Mobile phase M ix 15 volumes of tetrahydrqfuran R,
2,3,4,5-tetrahydro- IH -l -benzazepin- 1-yl] acetate.
25 volumes of methanol R and 60 volumes of a 2.0 g/L
PhEur solution o f citric add R.
Flow rate 0.8 mL/min.
Detection Spectrophotom eter at 273 nm.
Injection 20 jiL.
Run time Twice the retention time of bendroflumethiazide.
1-248 Benorilate 2016
ov
— mobile phase B: acetonitrUe R; •Y-NH
Time
(min)
Mobile phase A
(per cent V/V)
Mobile p h u t B
(per cent V/V)
C f 't )
0 -15
15-20
100 ->60
60
0*40
40
o ^ ~ 'r
20-25 100 0 B . 1- [ 1- [4-(2-fluorophenyl)-4-oxobutyl]piperidin-4-yI] -1,3-
d£hydro-2//-benzim idazol- 2 -one,
ov
V -N H OH
Content
98.5 per cent to 101.0 per cent (anhydrous substance). Flow rate 1.3 m U m in.
C H A R A C T ER S Detection Spectrophotometer at 210 nm.
Appearance Injection 5 pL.
White or yellowish-white or orange-white, crystalline powder.
Identification of impurities Use die chromatogram supplied
S olubility with benserazide for peak identification CRS and die
Freely soluble in water, very slightly soluble in anhydrous chromatogram obtained with reference solution (c) to identify
ethanol, practically insoluble in acetone. the peaks due to impurities A, B and C; doubling of die peak
It shows polymorphism (5.9). due to impurity C , related to separation o f the (£Z)-isomers,
may be observed.
ID E N T IF IC A T IO N
A. Infrared absorption spectrophotometry (2.2.24). Relative retention W ith reference to benserazide (retention
time = about 9 min): impurity A = about 0.6;
Comparison benserazide hydrochloride CRS.
impurity C = about 1.2; impurity B = about 1.5.
If die spectra obtained show differences, dissolve the System suitability: reference solution (a):
substance to be examined and die reference substance — resolution: minimum 5.0 between die peaks due to
separately in hot methanol R, evaporate to dryness and record benserazide and impurity C; use the 1st peak of
new spectra using the residues. impurity C if 2 peaks occur.
B. Solution S (see Tests) gives reaction (b) of chlorides Limits:
(2.3.1). — correction factor, for die calculation o f content, multiply the
TESTS peak area o f impurity B by 0.7;
Solution S — impurity A: not more than die area o f the corresponding
Dissolve 1.0 g in carbon dioxide-free water R and dilute to peak in the chromatogram obtained with reference
100 m L with die same solvent solution (a) (0.5 per cent);
— impurity B: not more than die area of the peak due to
Appearance of solution
benserazide in the chromatogram obtained with reference
Solution S is clear (2.2.1) and not more intensely coloured
solution (a) (0.5 per cent);
than reference solution BY6 (2.2.2, Method II).
— impurity C: not more than die area o f the corresponding
p H (2.2.5) peak or pair of peaks in the chromatogram obtained with
4.0 to 5.0 for solution S. reference solution (a) (0.5 per cent);
R elated su b stan ces — unspecified impurities: for each impurity, not more than the
Liquid chromatography (2.2.29). area o f the peak due to benserazide in die chromatogram
AM sohaions must be irgected immediatdy or stored at 4 °C. obtained with reference solution (b) (0.10 per cent);
— sum of impurities other than A: n o t more than twice the
Test solution Dissolve 0.100 g of die substance to be
area of die peak due to benserazide in die chromatogram
examined in methanol R2 and dilute to 50.0 m L with the
ob tain ed with reference solution (a) (1.0 per cent);
same solvent
— disregard Umar. 0.5 times die area of the peak due to
Reference solution (a) Dissolve 5.0 m g o f benserazide benserazide in the chromatogram obtained with reference
impurity A CRS, 5.0 m g of benserazide impurity C CRS angl solution (b) (0.05 per cent).
5.0 mg of benserazide hydrochloride CRS in methanol R2 and
dilute to 50.0 m L with the same solvent Dilute 5.0 m L of H eavy m e ta ls (2.4.8)
this solution to 50.0 m L with methanol R2. Maximum 20 ppm.
Reference solution (b) Dilute 2.0 m L of reference solution (a) 1.0 g complies with test C. Prepare the reference solution
to 10.0 m L with methanol R2. using 2 m L o f lead standard solution (10 ppm Pb) R
Reference solution (c) Dissolve 5 m g of benserazide for peak W a te r (2.5.12)
identification CRS (containing impurities A, B and C) in Maximum 1.0 per cent, determined on 0.500 g.
methanol R2 and dilute to 5.0 mL with the same solvent S u lfated a sh (2.4.14)
Column: Maximum 0.1 per cen^ determined on 1.0 g.
— size: I = 0.25 m, 0 = 4 mm; ASSAY
— stationary phase: octylsQyl silica gelfor chromatography R In order to avoid overheating during the titration, mix thoroughly
(5 Jim); throughout and stop the titration immediately after the end-point
— temperature. 30 °C.
has been reached
Mobile phase:
Dissolve 0.250 g in 5 m L o f anhydrous formic add R
— mobile phase A: dissolve 2.2 g of sodium heptanestdfbnate
Add 70 m L of anhydrous acetic add R Titrate immediately
monohydrate R and 6.8 g of potassium dihydrogen
with 0.1 M perchloric add, determining the end-point
phosphate R in 900 m L o f water R, add 50 m L of
potentiometrically (2.2.20).
methanol R2 and adjust to pH 3.5 with phosphoric add R;
— mobile phase B: dissolve 2.2 g of sodium heptanesulfonate 1 m L of 0.1 M perchloric add is equivalent to 29.37 mg of
monohydrate R and 6.8 g of potassium dihydrogen C 10H 16CIN 3O 5.
phosphate R in 500 m L o f water R, adjust to p H 3.5 with ST O R A G E
phosphoric add R and add 500 m L of methanol R2; Protected from light.
1-252 Bentonite 2016
8001-54-5
CrHfiO 106.1 100-52-7
Action and use
Action and use
Antiseptic.
Flavour.
PhEir____________
DEFINITION
Benzaldehyde c o n tains not less than 98.0% w/w and not DEFINITION
more than 100.5% w/w of G/H^O. M ixture of alkylbenzyldrmethylaxnmonium chlorides, the
alkyl groups mainly having chain lengths o f C 12, C J4 and
CHARACTERISTICS
A clear, colourless liquid.
c16.
Content
Slightly soluble in water, miscible with ethanol (96%) and
95.0 per cent to 104.0 p er cent of
with ether.
alkylbenzyldimethylammonium chlorides (anhydrous
TESTS substance) calculated using the average relative molecular
Refractive index m ass (see Tests).
1.544 to 1.546, Appendix V E. CHARACTERS
Weight per mL Appearance
1.043 to 1.049 g, Appendix V G. W hite or yellowish-white powder or gelatinous, yellowish-
Free add white fragments, hygroscopic. On heating it forms a d ear
N ot more than 1.0% w/v, calculated as benzoic ad d , m olten mass.
CyHfiO^ w hen determ ined by the following method. Solubility
T o 10 m L add 2 0 m L of ethanol (96%) previously Very soluble in water and in ethanol (96 p er cent).
neutralised to phenolphthalein solution R1 and titrate with A n aqueous solution froths copiously when shaken.
0 .1 m sodium hydroxide F 5 using phenolphthalein solution R1 as
IDENTIFICATION
indicator. E ach m l , of 0 .1 m sodium hydroxide P'S is equivalent
to 12.21 m g o f C 7H 60 2.
First identification B, E
Second identification A, C, D, E
Chlorinated compounds
N o t more than 0.05% w/v, calculated as Cl, when A . Ultraviolet and visible absorption spectrophotometry
determined by the following method. T o 5 m L add 50 m L of (2.2.25).
isoamyl alcohol and 3 g o f sodium and boil under a reflux Test solution Dissolve 80 m g in water R and dilute to
condenser for 1 hour. Cool, add 50 m L o f water and 15 m l, 100.0 m L with the same solvent.
of nitric acid, cool, add 5 m L o f 0.1m silver nitrate FS, shake Spectral range 220-350 nm .
and titrate the excess silver nitrate with 0 . 1m ammonium Absorption maxima At 257 nm, 263 nm and 269 nm.
thiocyanate VS using ammonium iron(III) sulfate solution R2 as
indicator. Repeat the procedure without the substance being
Shoulder A t about 250 nm .
examined. T h e difference between die titrations represents B. Examine the chromatograms obtained in the test for
the am ount o f silver nitrate required. Each m L o f 0 . 1m silver average rdative molecular mass and ratio o f alkyl
nitrate F51is equivalent to 3 .5 4 5 mg o f CL components.
Results T he prindpal peaks in the chromatogram obtained
ASSAY
w ith the test solution are similar in retention time to the
Carry out the m ethod for determination of aldehydes,
p rin d p al peaks in the chromatogram obtained with the
Appendix X K, using 0.5 g. Each m L o f 0.5m potassium
reference solution.
hydroxide in ethanol (60%) FiS is equivalent to 53.06 mg o f
CrHfiO. C . T o 2 m L of solution S (see Tests) add 0.1 m L o f glacial
acetic acid R and, dropwise, 1 m L o f sodium tetraphenylborate
STORAGE solution R. A white predpitate is formed. Filter. Dissolve the
Benzaldehyde should be kept in a well-filled container, p redpitate in a mixture o f 1 m L o f acetone R and 5 m L o f
protected from light and stored at a tem perature not ethanol (96 per cent) R, heating to not m ore than 70 °C.
exceeding 15°. A dd water R dropwise to the warm solution until a slight
opalescence forms. H eat gently until the solution is clear and
allow to cool. W hite crystals separate. Filter, wash with
3 quantities, each of 10 m T, of water R and dry in vacuo over
diphosphorus pentoxide R o r anhydrous silica gel R a t a
tem perature not exceeding 50 °C. The crystals melt (2.2.14)
at 127 °C to 133 °C.
D . T o 5 m L o f dilute sodium hydroxide solution R add 0.1 m L
o f bromophenol blue solution R1 and 5 m L o f methylene
chloride R and shake. T he methylene chloride layer is
1-254 Benzalkonium Chloride 2016
colourless. Add 0.1 m L of solution S and shake. Calculate the percentage of each homologue, using the
T he methylene chloride layer becomes blue. following expression:
E. T o 2 m L o f solution S add 1 m L o f dilute nitric acid R.
A white precipitate is formed which dissolves on the addition
of 5 m L of ethanol (96 per cent) R. T h e solution gives
reaction (a) of chlorides (2.3.1).
100(I)
TESTS C = product of the relative molecular mass o f the given
S o lu tio n S homologue and the area of the corresponding peak
Dissolve 1.0 g in carbon dioxide-free water R and dilute to in the chromatogram obtained with the test
100 m L with the same solvent. solution;
A p p e a ra n c e o f so lu tio n D = sum o f the C values for all homologues quantified.
Solution S is clear (2.2.1) and not more intensely coloured Limits:
than reference solution Y 6 (2.2.2, MethodII). — C1 2 homologue: minimum 40 per cent;
A cid ity o r alk alin ity — C14 homologue: minimum 20 p er cent;
T o 50 m L of solution S add 0.1 m L o f bromocresolpurple — sum of C 1 2 and C14 homologues: minim um 70 per cent.
solution R. N ot more than 0.1 m L of 0.1 M hydrochloric add C 1 2 and C14 homologues
or 0.1 M sodium hydroxide is required to change the colour of Im p u ritie s A , B a n d C
the indicator. Liquid chromatography (2.2.29). Prepare the solutions
A v erag e re la tiv e m o le c u la r m a s s a n d ra tio o f alkyl immediately before use.
co m p o n e n ts Test solution Dissolve 0.50 g o f the substance to be examined
L iquid chromatography (2.2.29). in methanol R1 and dilute to 10.0 m L with the same solvent.
Test solution Dissolve 0.400 g o f the substance to be Reference solution (a) Dissolve 25.0 mg of benzyl alcohol CRS
examined in water R and dilute to 100.0 m L with the same (impurity A) in methanol R1 and dilute to 100.0 m L with the
solvent. same solvent.
Reference solution Dissolve the contents o f a vial of Reference solution (b) Dissolve 75.0 m g of benzaldehyde CRS
benzalkonium chloridefor system suitability CRS in 5.0 m L o f (impurity B) in methanol R1 and dilute to 100.0 m L with the
water R. same solvent. Dilute 1.0 m L o f this solution to 10.0 m L with
Column: methanol R l.
— sizer. I = 0.25 m , 0 = 4.6 mm; Reference solution (c) Dilute 1.0 m L o f reference solution (a)
— stationary phase: end-capped nitrUe silica gelfor to 10.0 m L with methanol R l.
chromatography R (5 pm). Column:
Mobile phase Mix 45 volumes o f acetonitrUe R and 55 volumes — size. I = 0.15 m, 0 = 4.6 mm;
of a 13.6 g/L solution o f sodium acetate R previously adjusted — stationary phase, end-capped octadecylsUyl silica gel for
to p H 5.0 with glacial acetic add R. chromatography R (5 jam);
Flow rate 2.0 mL/min. — temperature: 30 °C.
Detection Spectrophotom eter at 254 nm. Mobile phase.
— mobile phase A: dissolve 1.09 g o f sodium hexanesulfimate R
Injection 10 (iL.
and 6.9 g of sodium dihydrogen phosphate monohydrate R in
Identification of homologues Use the chromatogram supplied water R; adjust to p H 3.5 with phosphoric add R and
with benzalkonium chloride for system suitability CRS and the dilute to 1000.0 m L with the same solvent;
chromatogram obtained with die reference solution to — mobile phase B: m ethanol R l ;
identify the peaks due to Q jj, C 14 and C i6.
Relative retention W ith reference to C 12 homologue (retention
Time Mobile phase A Nobile phase B
time = about 6 min): C 14 homologue = about 1.3;
(min) (per cent V/V) (per cent V/V)
C 16 homologue = about 1.7.
0-10 80 20
System suitability: reference solution:
— resolution: minim um 1.5 between the peaks due to the C i 2 10-14 80 —> 50 2 0 -* 5 0
and C 14 homologues. 14-35 50 50
Calculate the average relative molecular mass o f the sample 35 -3 6 50 —> 20 5 0 -» 8 0
by summing the products for each homologue, using the
following expression: 3 6-55 20 80
3 6 -5 5 20 80
2016 Benzathine Benzylpenicillin 1-257
^ ^ nh2
,CH3S 03 H
OCHPh2
D E F IN IT IO N
Benzatropine Mesilate is (li?,3i?,55)-3-benzhydryloxytropane
methanesulfonate. It contains not less than 98.0% and not
m ore than 100.5% o f C 2iH 25N 0 ,C H 403S, calculated with
reference to the dried substance.
C H A R A C T E R IS T IC S
A white, crystalline powder. It melts at about 144°.
Very soluble in water, fredy soluble in ethanol (96%) \
practically insoluble in ether.
C. benzylpenicilloic ad d s benzathide, ID E N T IF IC A T IO N
A. D ry the substance at 105° for 3 hours. The infrared
absorption spectrum, Appendix II A, is concordant with the
,c o 2h reference spectrum of benzatropine mesilate (RS 026).
ch 3 B. T h e light absorption, Appendix II B, in the range 230 to
ch 3 350 nm of a 0.1% w/v solution in 2m hydrochloric acid
H02C I H exhibits two maxima, at 253 and 258 nm . T he absorbance at
n
253 nm is about 0.96 and at 258 n m is about 1.1.
C. Dissolve 10 m g in 2 m L of water, pour into 5 m L o f hot
D . (35,7i?,7aR)-5-benzyl-2,2-dimethyl-2,3,7,7a-
picric arid solution R1 and allow to c o o l T he melting point of
tetrahydroimidazo [5,1 -b]thiazole-3,7~dicarboxylic a d d
the predpitate, after drying at 105°, is about 185°,
(penillic a d d o f benzylpenicillin),
Appendix V A.
TESTS
I.COzH
T ro p in e
h n - A .c h 3
C arry out the m ethod for thin-layer chromatography,
Appendix HI A, using silica gel G as the coating substance
o co 2h
and a mixture of 75 volumes of ethanol (96%) and
15 volumes o f 13. 5m ammonia as the mobile phase. Apply
separatdy to the plate 10 |iL of each o f two solutions in
E. (45)-2-[caiboxy[(phenylacetyl)amino]methyl]-5,5- acetone containing (1) 4.0% w/v of the substance being
dimethylthiazoüdine-4-carboxylic a d d (penicilloic ad d s of examined and (2) 0.020% w/v o f tropine. After removal o f the
benzylpenicillin), plate, allow it to dry in air and spray with sodium
iodobismuthate solution and then with a 0.4% w/v solution of
sulfuric acid. Any spot corresponding to tropine in the
1-260 Benzbromarone 2016
H eav y m e ta ls (2.4.8) D E F IN IT IO N
M axim um 20 ppm . N -B enzyl-N ^-dim ethyl-2- [2- [4-( 1,1,3,3-
0.5 g complies with test C. Prepare the reference solution tetramethylbutyl)phenoxy] ethoxy] ethanaminium chloride.
using 1 m L o f lead standard solution (10 ppm Pb) R. C o n te n t
97.0 per cent to 103.0 per cent (dried substance).
1-262 Benzocaine 2016
CHARACTERS ASSAY
A p p e a ra n c e Dissolve 2.000 g in water R and dilute to 100.0 m L with the
W hite or yellowish-white powder. same solvent. Transfer 25.0 m L o f the solution to a
S o lubility separating funnel, add 10 m L of a 4 g/L solution o f sodium
Very soluble in water and in ethanol (96 per cent), freely hydroxide R, 10.0 m L of a freshly prepared 50 g/L solution of
soluble in methylene chloride. potassium iodide R and 25 m L o f methylene chloride R. Shake
vigorously, allow to separate and discard the lower layer.
An aqueous solution froths copiously when shaken.
Shake the upper layer with 3 quantities, each of 10 mT, of
ID E N T IF IC A T IO N methylene chloride R and discard the lower layers. T o the
A. Melting point (2.2.14): 158 °C to 164 °C, after drying at upper layer add 40 m L o f hydrochloric add R, allow to cool
105 °C for 4 h. and titrate with 0.05 M potassium iodate until the deep brown
B. Thin-layer chromatography (2.2.27). colour is almost discharged. Add 4 m L o f methylene chloride R
Test solution Dissolve 25 mg o f the substance to be examined and continue the titration, shaking vigorously, until the lower
layer is no longer brown. C any ou t a blank titration using a
in water R and dilute to 5 m L with the same solvent.
mixture of 10.0 m L o f a freshly prepared 50 g/L solution of
Reference solution Dissolve 25 m g of benzethonium chloride CRS potassium iodide R, 20 m L o f water R and 40 m L of
in water R and dilute to 5 m L with the same solvent.
hydrochloric acid R.
Plate TLC silica gel F2 5 4 plate R. 1 m L of 0.05 Mpotassium iodate is equivalent to 44.81 m g of
Mobile phase glacial acetic acid R, water R, methanol R C 27H 42CINO 2.
(5:5:100 V/V/V).
STORAGE
Application 20 pL.
Protected from light.
Development Over a path of 12 cm.
PhEur
Drying In a current of warm air.
Detection Examine in ultraviolet light at 254 nm.
Results T he principal spot in the chromatogram obtained with
the test solution is similar in position and size to the principal ★ ★
spot in the chromatogram obtained with the reference Benzocaine ★ ★
solution. (Ph. Eur. monograph 0011) **★**
C. T o 5 m L of dilute sodium hydroxide solution R add 0.1 m L
of bromophenol blue solution R l and 5 m L o f methylene
chloride R and shake. T h e lower layer is colourless.
Add 0.1 m L o f solution S (see Tests) and shake. A blue 'CH,
colour develops in the lower layer.
h2n
D. T o 2 m L o f solution S add 1 m L of dilute nitric acid R.
A white precipitate is formed which dissolves upon addition
of 5 m L o f ethanol (96 per cent) R. T he solution gives CçHuNOa 165.2 94-09-7
reaction (a) o f chlorides (2.3.1).
A c tio n a n d u se
TESTS Local anaesthetic.
S o lu tio n S
Dissolve 5.0 g in carbon dioxide-free water R and dilute to PhEur.
50 m L with the same solvent. DEFINITION
Appearance o f solution Ethyl 4-aminobenzoate.
Solution S is clear (2.2.1) and not more intensely coloured Content
than reference solution Y6 (2.2.2, Method II). 99.0 per cent to 101.0 per cent (dried substance).
Acidity or alkalinity
CHARACTERS
T o 25 m L o f solution S add 0.1 m L of phenolphthalein
solution R. T he solution is colourless. Add 0.3 m L o f 0.01 M Appearance
W hite or almost white, crystalline powder or colourless
sodium hydroxide. T h e solution is pink. Add 0.1 m L of methyl
crystals.
red solution R and 0.5 m L o f 0.01 M hydrochloric acid
T he solution is orange-red. Solubility
Very slightly soluble in water, freely soluble in ethanol
V olatile b a se s a n d sa lts o f v o latile b ase s (2.4.1,
Method B) (96 per cent).
M aximum 50 ppm , determined on 0.20 g. It shows polymorphism (5.9).
Prepare the standard using 0.1 m L of ammonium standard IDENTIFICATION
solution (100 ppm N H J R. Replace heavy magnesium oxide Infrared absorption spectrophotometry (2.2.24).
by 2.0 m L o f strong sodium hydroxide solution R. Comparison benzocaine CRS.
L oss o n d ry in g (2.2.32) If the spectra obtained show differences, dissolve the
M aximum 5.0 per cent, determined on 1.000 g by drying in substance to be examined and the reference substance
an oven at 105 °C for 4 h. separately in anhydrous ethanol R, evaporate to dryness and
S u lfa te d a s h (2.4.14) record new spectra using the residues.
Maximum 0.1 per cent, determined on 1.0 g. TESTS
Related substances
Liquid chromatography (2.2.29).
2016 Benzocaine 1-263
L oss o n d ry in g (2.2.32)
M aximum 0.5 per cent, determined on 1.000 g by drying
in vacuo.
S u lfa te d a s h (2.4.14)
M aximum 0.1 per cent, determined on 1.0 g.
G. 4-aminobenzoic acid,
A SSA Y
Carry out the determination of primary aromatic amino- o
nitrogen (2.5.5), using 0.400 g dissolved in a mixture of
25 m L of hydrochloric acid R and 50 m L o f water R.
1 m L o f 0.1 M sodium nitrite is equivalent to 16.52 mg
o fG jH n N O z .
ST O R A G E H . methyl 4-aminobenzoate.
Protected from light. __________________________________________________________PhEur
1-264 Benzoic Acid 2016
— water, m inim um 20.0 per cent. phase. Dilute 1.0 m L of the solution to 10.0 mT. with the
mobile phase.
CHARACTERS
Appearance Reference solution (c) Dissolve 50.0 mg of ethyl benzoate R in
White o r alm ost white, amorphous or granular powder. the mobile phase and dilute to 100.0 m L with the mobile
phase. Dilute 1.0 m L o f the solution to 100.0 m L with the
S o lu b ility mobile phase.
Practically insoluble in water, soluble in acetone, soluble in
methylene chloride with the separation of water, slightly
Reference solution (d) Dissolve 50.0 mg of benzaldehyde R in
the mobile phase and dilute to 100.0 m L with the mobile
soluble in ethanol (96 per cent).
phase. Dilute 1.0 m L o f the solution to 100.0 m L with the
It loses w ater rapidly on exposure to air with a risk of
mobile phase.
explosion.
Reference solution (e) Dissolve 30.0 mg of benzoic add R and
Mix the entire sample thoroughly before carrying out the following 30.0 m g of benzaldehyde R in the mobile phase and dilute to
tests. 100.0 m L with the mobile phase. Dilute 1.0 m L of the
ID E N T IF IC A T IO N solution to 10.0 m L with the mobile phase.
First identification B Column:
Second identification A y C, D. — size: I = 0.25 m , 0 = 4.6 mm;
A. U ltraviolet and visible absorption spectrophotometry — stationary phase: octadecylsüyl silica gel for chromatography R
(10 |im).
(2.2.25).
Solution A Dissolve 80.0 mg in ethanol (96 per cent) R and Mobile phase glacial acetic acid R, acetonitrüe R, water R
dilute to 100.0 m L with the same solvent. Dilute 10.0 m L o f (1:500:500 V/V/V).
the solution to 100.0 m L with ethanol (96 per cent) R. Flow rate 1 mL/min.
Solution B D ilute 10.0 m L of solution A to 100.0 m L with Detection Spectrophotometer at 235 nm.
ethanol (96 per cent) R. Injection 20 |iL loop injector.
Spectral ranges 250-300 n m for solution A; 220-250 nm for Run time 2 times the retention time of dibenzoyl peroxide.
solution 6 . Relative retention W ith reference to dibenzoyl peroxide
Absorption maxima A t 274 nm for solution A; at 235 nm for (retention time = about 28.4 min): impurity B = about 0.15;
solution B. impurity A = about 0.2; impurity C = about 0.4.
Shoulder A t about 282 nm for solution A. System suitabüity: reference solution (e):
Absorbance ratio A 2 3 5 /A 2 7 4 = 1.17 to 1.21. — resolution: minimum 6 between the peaks due to benzoic
acid and benzaldehyde.
B. Infrared absorption spectrophotometry (2.2.24).
Limits:
Comparison Ph. Eur. reference spectrum of hydrous benzoyl
— impurity A: n o t m ore than the area of the principal peak
peroxide.
in the chromatogram obtained with reference solution (d)
C. Dissolve about 25 m g in 2 m L o f acetone R. Add 1 m L o f (0.25 per cent);
a 10 g/L solution o f diethylphenylenediamme sulfate R and mix. — impurity B: not more than the area of the principal peak
A red colour develops which quickly darkens and becomes in the chromatogram obtained with reference solution (b)
dark violet within 5 mm. (1.5 per cent);
D . T o 1 g add 5 mT. of ethanol (96 per cent) R, 5 m L of — impurûy C: not more than the area of the principal peak
dilute sodium hydroxide solution R and 10 m L of water R Boil in the chromatogram obtained with reference solution (c)
the m ix ture under reflux for 20 min. Cool. T he solution (0.25 per cent);
gives reaction (c) o f benzoates (2.3.1). — unspecified impurities: for each impurity, not m ore than the
TESTS area of the principal peak in the chromatogram obtained
A cid ity with reference solution (a) (0.10 per cent);
Dissolve a quantity o f the substance to be examined — disregard limit:. 0.2 times the area of the principal peak in
containing the equivalent o f 1.0 g o f dibenzoyl peroxide in the chromatogram obtained with reference solution (a)
25 m L o f acetone R, add 75 m L of water R and filter. Wash (0.02 per cent).
the residue with two quantities, each of 10 mL, o f water R. Chlorides (2A. 4)
Com bine the filtrate and the washings and add 0.25 m L of M aximum 0.4 per cent.
phenolphthalein solution R l. N o t more than 1.25 m L of 0.1 M Dissolve a quantity of the substance to be ex a m in ed
sodium hydroxide is required to change the colour of the containing the equivalent of 0.5 g of dibenzoyl peroxide in
indicator. Carry out a blank test. 15 m L of acetone R. Add, while stirring, 50 m L o f 0.05 M
Related substances nitric acid. Allow to stand for 10 min and filter. W ash the
Liquid chrom atography (2.2.29). Prepare the solutions residue with 2 quantities, each of 10 mL, of 0.05 M nitric
immediately before use. acid. Combine the filtrate and the washings and dilute to
Test solution Dissolve a quantity of the substance to be 100 m L with 0.05 M nitric acid. Dilute 2.5 m L of the
examined containing the equivalent of 0.10 g of dibenzoyl solution to 15.0 m L w ith water R.
peroxide in acetonitrüe R and dilute to 50 m L with the same A SSAY
solvent. Solution (a) Dissolve 2.500 g immediately before use in
Reference solution (a) Dilute 1.0 m L of the test solution to 75 m L of dimethylformamide R and dilute to 100.0 m L with
100.0 m L with acetonitrüe R. D ilute 1.0 m L of this solution the same solvent.
to 10.0 m L with acetonitrüe R. Dibenzoyl peroxide
Reference solution (b) Dissolve 30.0 m g of benzoic acid R in T o 5.0 m L of solution (a) add 20 m L of acetone R and 3 m L
the m obile phase and dilute to 100.0 m L with the m obile, o f a 500 g/L solution o f potassium iodide R and mix. Allow to
1-266 Benzydamine Hydrochloride 2016
o 1 - 3-dimethylaminopropyl 2-benzylaminobenzoate
hydrochloride BPCRS (impurity A) and 0.00125% w/v of
A. R = H: benzaldehyde, 3-(l,5dibenzyl-lH-mdazole-3-yl)oxypropyldimethylamirie
hydrochloride BPCRS (impurity B). Solution (3) contains
B. R = OH: benzoic acid,
0.00025% w/v o f 1-benzyl-lH-mdazol-3-ol BPCRS
C. R = 0 -C H 2-C H 3: ethyl benzoate. (impurity C). Solution (4) contains 0.00025% w/v o f the
PhEur substance being examined. Solution (5) contains equal
volumes o f solutions (1), (2) and (3).
T h e chromatographic procedure may be carried out using a
stainless steel colum n (25 cm x 4.6 mm ) packed with end-
capped octadecylsUyl silica gelfor chromatography (5 fun)
Benzydamine Hydrochloride (Suplex pK B-100 is suitable), fitted with a stainless steel
guard colum n (2 cm x 4.6 mm ) packed with the same
m aterial U se as the initial mobile phase with a flow rate of
1.5 m L per m inute, a m ixture o f 50 volumes o f solution A
and 50 volumes o f methanol. Solution A contains
0.01m potassium dihydrogen orthophosphate and 0.005m sodium
octyl sulfate in water, adjusted to p H 3.0 ± 0 . 1 with
orthophosphoric add Carry out a linear gradient elution
increasing the percentage o f methanol to 70% over
20 m inutes from die m om ent of injection and then
C19H23N3OJHCI 132-69-4 decreasing the percentage o f methanol to 50% over 2 minutes
and maintain the final mobile phase until the completion of
Action and use th at run. U se a detection wavelength o f 320 nm .
Cyclo-oxygenase inhibitor; analgesic; anti-inflam m atory
Inject 20 jiL of solution (5) and modify the rate o f change o f
Preparations the mobile phase, if necessary, to obtain a retention time of
Benzydamine Cream about 10 m inutes for the substance being examined.
Benzydamine M outhwash
Benzydamine Oromucosal Spray
2016 Benzyl Alcohol 1-267
to 25.0 m L with the test solution. A dd 1.0 m L o f this — disregard litrtir. 0.01 times the area o f the peak due to
solution to a mixture o f 2.0 m L o f standard solution (a) and ethylbenzene in the chrom atogram obtained with
2.0 m L of standard solution (b) and dilute to 20.0 m L with reference solution (a) corrected if necessary as
the test solution. described above (0.0001 p er cent).
Column: Benzyl alcohol intendedfor parenteral administration
— material: fused silica; Injection W ithout air-plug, 0.1 nL o f the test solution and
— size: / = 30 m, 0 = 0.32 m m ; reference solution (b).
— stationary phase: macrogol 20 000 R (film thickness
Relative retention W ith reference to benzyl alcohol (retention
0.5 nm).
time = about 26 min): ethylbenzene = about 0.28;
Carrier gas helium for chromatography R. dicyclohexyl = about 0.59; impurity A = about 0.68;
Linear velocity 25 cm/s. impurity B = about 0.71.
Temperature: System suitability: reference solution (b):
— resolution: m inim um 3.0 between the peaks due to
Time Temperature
impurities A and B.
(min) CC) If any peaks in the chrom atogram obtained with the test
Column 0 - 34 50*220 solution have die same retention times as the peaks due to
ethyl benzene o r dicyclohexyl, substract the areas o f any such
3 4 -6 9 220
peaks from die peak areas at these retention times in the
Injection port 200 chromatograms obtained with reference solutions (a) o r (b)
Detector 310 (corrected peak areas of ethyl benzene and dicyclohexyl).
Any such peaks in the chrom atogram obtained with the test
solution are to be included in the assessments for the sum of
Detection Flam e ionisation. other peaks.
Benzyl alcohol not intended for parenteral administration Limits:
Injection W ithout air-plug, 0.1 jtL o f the test solution and — impurity A: n o t m ore than the difference between the
reference solution (a). area o f die peak due to impurity A in the
Relative retention W ith reference to benzyl alcohol (retention chrom atogram obtained w ith reference solution (b)
time = about 26 min): ethylbenzene = about 0.28; and the area o f the peak due to impurity A in the
dicyclohexyl = about 0.59; im purity A = about 0.68; chrom atogram obtained w ith the test solution
impurity B = about 0.71. (0.05 per cent);
— impurity B: n o t m ore than the difference between the
System suitability: reference solution (a):
area o f die peak due to impurity B in the
— resolution: m inim um 3.0 between the peaks due to
chrom atogram obtained with reference solution (b)
impurities A and B.
and the area o f the peak due to impurity B in the
If any peaks in the chrom atogram obtained with the test chrom atogram obtained with the test solution
solution have the sam e retention tim e as the peaks due to (0.10 p er cent);
ethyl benzene or dicyclohexyl, substract the areas o f any such — sum of other peaks with a relative retention less than that
peaks from the peak areas at these retention times in the of benzyl alcohol: n o t m ore than twice the area of the
chromatograms obtained with reference solutions (a) o r (b) peak due to ethylbenzene in the chromatogram
(corrected peak areas o f ethyl benzene and dicyclohexyl). obtained with reference solution (b) corrected if
Any such peaks in the chrom atogram obtained w ith the test necessary as described above (0.02 p er cent);
solution are to be included in die assessments for the sum of — sum of peaks with a relative retention greater than that of
other peaks. benzyl alcohol: n o t m ore than the area o f the peak due
Limits: to dicyclohexyl in the chrom atogram obtained with
— impurity A: n o t m ore than the difference betw een the reference solution (b) corrected if necessary as
area o f the peak due to impurity A in the described above (0.2 per cent);
chrom atogram obtained with reference solution (a) — disregard limit: 0.01 times the area o f the peak due to
and the area o f the peak due to impurity A in the ethylbenzene in the chrom atogram obtained with
chrom atogram obtained w ith the test solution reference solution (b) corrected if necessary as
(0.15 per cent); described above (0.0001 per cent).
— impurity B: n o t m ore than the difference Jbetween the
Residue on evaporation
area o f the peak due to impurity B in the
M aximum 0.05 p er cent.
chrom atogram obtained with reference solution (a)
and the area o f the peak due to impurity B in the After ensuring th at th e substance to be examined complies
chrom atogram obtained with the test solution with the test for peroxide value, evaporate 10.0 g to dryness
(0.10 p er cent); in a tared quartz o r porcelain crucible or platinum dish on a
— sum of other peaks with a relative retention less than that hot plate a t a tem perature n o t exceeding 200 °C. Ensure that
of benzyl alcohol: no t m ore than 4 times die area of the the substance to be examined does no t boil during
peak due to ethylbenzene in the chromatogram evaporation. D ry the residue on the ho t plate for 1 h and
obtained with reference solution (a) corrected if allow to cool in a desiccator. T h e residue weighs a maximum
necessary as described above (0.04 per cent); of 5 mg.
— sum of peaks with a relative retention greater than that of ASSAY
benzyl alcohol: n o t m ore than the area o f the peak due T o 0.900 g ( m g ) add 15.0 m L o f a freshly prepared mixture
to dicyclohexyl in the chrom atogram obtained with of 1 volume o f acetic anhydride R and 7 volumes o f anhydrous
reference solution (a) corrected if necessary as pyridine R and heat u n d er a reflux condenser on a boiling
described above (0.3 p er cent);
2016 Benzyl Hydroxybenzoate 1-269
PhEti.
D E F IN IT IO N
Phenylmethyl benzoate. Benzyl Hydroxybenzoate
C o n te n t Benzylparaben
99.0 per cent to 100.5 per c e n t
CHARACTERS
A p p e a ra n c e
Colourless or almost colourless crystals or colourless or
almost colourless, oily liquid.
S o lubility
Practically insoluble in water, miscible with ethanol
C 14H 12O3 228.3 94-18-8
(96 per cent), with methylene chloride and with fatty and
essential oils. A ctio n a n d u se
Eb: about 320 °C. Antimicrobial preservative.
1-270 Benzylpenicillin Potassium 2016
D E F IN IT IO N
Benzylpenicillin Potassium *****
Benzyl Hydroxybenzoate is benzyl 4-hydroxybenzoate.
It contains not less than 99.0% and n o t more than 101.0% (Ph. Eur. monograph 0113) *
of C 14H 12O 3.
0 H1 C O jK
C H A R A C T E R IS T IC S
Y - N '''\.C H 3
A white to creamy white, crystalline powder.
Practically insoluble in water, freely soluble in ethanol (96%) n - - |—4 ^s ch3
H H
and in ether. It dissolves in solutions o f alkali hydroxides.
ID E N T IF IC A T IO N
A. T h e infrared absorption spectrum, Appendix II A, is Ci6H 17KN 20 4S 372.5 113-98-4
concordant with the reference spectrum of benzyl
hydroxybenzoate (RS 028). A c tio n a n d u se
B. T h e light absorption, Appendix II B, in the range 230 to Penicillin antibacterial.
350 nm of a 0.001% w/v solution in ethanol (96%) exhibits a P r e p a r a tio n
m axim um only at 260 nm . T h e absorbance at the maximum Benzylpenicillin Injection
at 260 nm is about 0.76.
C. Dissolve 0.1 g in 2 m L of ethanol (96%), boil and add
PhEur___________________________________________________
0.5 m L of nitric add solution of mercury. A precipitate is D E F IN IT IO N
produced slowly and the supernatant liquid becomes red. Potassium (2S,5Ä,6.R)-3,3-dimethyl-7-oxo-6-
D. Melting point, about 112°, Appendix V A. [(phenylacetyl) amino] -4-thia-1-azabicydo [3.2.0] heptane-2-
TESTS carboxylate.
A cid ity Substance produced by the growth of certain strains of
Dissolve 0.2 g in 10 m L of ethanol (50%) previously PenicUHum notation or related organisms, or obtained by any
neutralised to methyl red solution and titrate with 0 .1 m sodium other means.
hydroxide KS using methyl red solution as indicator. N o t more C o n te n t
than 0.1 m L of 0. 1m sodium hydroxide VS is required to 96.0 per cent to 102.0 per cent (dried substance).
change the colour o f the solution.
CHA RACTERS
R e la te d su b sta n c e s
A p p e a ra n c e
Carry out the m ethod for thin-layer chromatography,
W hite or almost white, crystalline powder.
Appendix IH A, using a plate precoated with silica gel F 254j
the surface o f which has been modified with chemically- S o lu b ility
bonded octadecylsilyl groups (W hatm an K C18F plates are Very soluble in water, practically insoluble in fatty oils and in
suitable) and a mixture o f 70 volumes o f methanol, liquid paraffin.
30 volumes o f water and 1 volum e o f glacial acetic add as the ID E N T IF IC A T IO N
mobile phase. Apply separately to the plate 2 |iL o f each of First identification A , D
two solutions o f the substance being examined in acetone
Second identification B, C, D
containing (1) 1.0% w/v and (2) 0.010% w/v. After removal
of the plate, allow it to dry in air and examine under A. Infrared absorption spectrophotometry (2.2.24).
ultraviolet light (254 nm). Any secondary spot in the Comparison benzylpemdOin potassium CRS.
chrom atogram obtained with solution ( 1) is not more intense B. Thin-layer chromatography (2.2.27).
than the spot in the chrom atogram obtained with Test solution Dissolve 25 mg of the substance to be examined
solution. (2). in 5 m l, of water R.
Sulfated ash
Reference solution (a) Dissolve 25 m g of benzylpenidllin
N ot m ore than 0.1%, A ppendix IX A. potassium CRS in 5 m L of water R
A SSA Y Reference solution (b) Dissolve 25 m g of benzylpenicHlin
Gently boil 0.12 g under a reflux condenser with 20 m L of potassium CRS and 25 m g o f phenoxymethylpemdUin
2m sodium hydroxide for 30 minutes. Cool and extract with potassium CRS in 5 mT. o f water R
three 20-m L quantities o f 1,2-dichhroethane. Wash the
Plate TLC sUamsed silica gel plate R.
com bined extracts with 20 m L o f 0.1m sodium hydroxide and
. add the washings to the m ain aqueous phase, discarding the Mobile phase Mix 30 volumes of acetone R and 70 volumes of
organic layer. T o the aqueous solution add 25 m L of a 154 g/L solution of ammonium acetate R previously adjusted
0.0333m potassium bromate VS, 5 m L o f a 12.5% w/v solution to p H 5.0 with glacial acetic acid R.
of potassium bromide and 10 m L o f hydrochloric add and Application 1 |iL.
immediately stopper the flask. Shake for 15 minutes and Development Over a path o f 15 cm.
allow to stand for 15 minutes. A dd 25 m L o f dilute potassium Drying In air.
iodide solution and shake vigorously. T itrate the liberated
Detection Expose to iodine vapour until the spots appear and
iodine with 0 .1 m sodium thiosulfate VS using starch mucilage,
examine in daylight.
added towards the end of the titration, as indicator. Repeat
the operation w ithout the substance being examined. System suitability: reference solution (b):
T he difference between the titrations represents the am ount — the chromatogram shows 2 clearly separated spots.
of potassium brom ate required. T he volume of Results T he principal spot in the chromatogram obtained with
0 .0 3 3 3 m potassium bromate VS used is equivalent to h alf of the test solution is similar in position, colour and size to the
the volum e o f 0 . 1m sodium thiosulfate VS required for the principal spot in obtained with reference solution (a).
titration. Each m L o f 0 .0 3 3 3 m potassium bromate VS is C. Place about 2 m g in a test-tube about 150 m m long and
equivalent to 7.608 m g o f Q 4H 12O 3. 15 m m in diameter. Moisten with 0.05 m L o f water R and
2016 Benzylpenicillin Potassium 1-271
add 2 m L o f sulfuric acid-formaldehyde reagent R. Mix the If the mobile phase composition has been adjusted to achieve
contents of the tube by swirling; the solution is practically the required resolution, the adjusted composition will apply
colourless. Place the test-tube on a water-bath for 1 min; at time zero in the gradient and in the assay.
a reddish-brown colour develops. Flow rate 1.0 mL/min.
D . It gives reaction (a) of potassium (2.3.1). Detection Spectrophotometer at 225 nm.
TESTS Injection 20 jiL o f reference solutions (b) and (c) with
pH (2.2.3) isocratic elution at the initial mobile phase composition and
5.5 to 7.5. 20 nL o f test solution (b) according to the elution gradient
Dissolve 2.0 g in carbon dioxide-free water R and dilute to described under Mobile phase; inject water R as a blank
20 m L with the same solvent. according to the elution gradient described under Mobile
phase.
Specific optical rotation (2.2.7)
+ 270 to + 300 (dried substance). System suitability: reference solution (b):
— resolution: minim um 6.0 between the peaks due to
Dissolve 0.500 g in carbon dioxide-free water R and dilute to
impurity B and benzylpenicillin; if necessary, adjust the
25.0 m L with the same solvent.
ratio A:B o f the mobile phase.
Absorbance (2.2.25)
Limit:.
Dissolve 94.0 mg in water R and dilute to 50.0 m L with the
— any impurity: for each impurity, not more than the area of
same solvent. Measure the absorbance of the solution at
the principal peak in the chromatogram obtained with
325 nm , 280 nm and at the absorption maximum at 264 nm,
reference solution (c) (1 per cent).
diluting the solution, if necessary, for the measurement at
264 nm . T h e absorbances at 325 nm and 280 nm do not Loss on drying (2.2.32)
exceed 0.10 and that at the absorption maximum at 264 nm M axim um 1.0 per cent, determined on 1.000 g by drying in
is 0.80 to 0 .88, calculated on the basis o f the undiluted an oven at 105 °C.
(1.88 g/L) solution. Verify the resolution o f the apparatus Bacterial endotoxins (2.6.14, Method E)
(2.2.25); the ratio o f the absorbances is at least 1.7. Less than 0.16 IU/mg, if intended for use in the manufacture
Related substances o f parenteral preparations without a further appropriate
Liquid chromatography (2.2.29). Prepare the solutions procedure for the removal of bacterial endotoxins.
immediately before use. ASSAY
Test solution (a) Dissolve 50.0 mg o f the substance to be Liquid chromatography (2.2.29) as described in the test for
examined in water R and dilute to 50.0 m L with the same related substances with the following modifications.
solvent. Mobile phase Initial composition of the mixture of mobile
Test solution (b) Dissolve 80.0 mg of the substance to be phases A and B, adjusted where applicable.
examined in water R and dilute to 20.0 m L with the same Injection T est solution (a) and reference solution (a).
solvent.
Calculate the percentage content o f C 16H 17K N 20 4S by
Reference solution (a) Dissolve 50.0 mg o f benzylpenicillin multiplying the percentage content of benzylpenicillin sodium
sodium CRS in water R and dilute to 50.0 m L with the same by 1.045.
solvent.
STORAGE
Reference solution (b) Dissolve 10 mg of benzylpenicillin
sodium CRS and 10 mg of phenylacetic acid R (impurity B) in In an airtight container. If the substance is sterile, store in a
water R, then dilute to 50 m L with the same solvent. sterile, airtight, tamper-proof container.
S o lu b ility
W I c o 2h Very soluble in water, practically insoluble in fatty oils and in
k V cc h 3 liquid paraffin.
ch3 ID E N T IF IC A T IO N
V P *
HOî C ^ H First identification A, D
Second identification B, C, D
D. (3«Sj7i?,7ai?)-5-benzyl-2j2-dimethyl-2j3j7j7a- A. Infrared absorption spectrophotometry (2.2.24).
tetrahydroimidazo [5,1 -b]thiazole-3,7-dicarboxyiic a d d Comparison benzylpemdUin sodium CRS.
(penillic a d d o f benzylpenicillin).
B. Thin-layer chromatography (2.2.27).
PhEur
Results T h e prindpal spot in the chromatogram obtained with
the test solution is similar in position, colour and size to the
p rindpal spot in the chromatogram obtained with reference
solution (a).
*** C. Place about 2 m g in a test-tube about 150 m m long and
Benzylpenicillin Sodium ★ ★
★ ★ 15 m m in diameter. Moisten with 0.05 m L o f water R and
***** add 2 m L of sulfuric add-formaldehyde reagent R. Mix the
(Ph. Eur. monograph 0114)
contents o f the tube by swirling; the solution is practically
colourless. Place the test-tube on a water-bath for 1 min;
0 H COjNa
a reddish-brown colour devdops.
N ^ A _ch3
D . It gives reaction (a) of sodium (2.3.1).
TESTS
p H (2.2.3)
5.5 to 7.5.
C 16H 17N 2N a 0 4S 356.4 69-57-8 Dissolve 2.0 g in carbon dioxide-free water R and dilute to
20 m L with the same solvent.
A c tio n a n d u se S p ecific o p tic a l ro ta tio n (2.2.7)
Penicillin antibacterial. + 285 to + 310 (dried substance).
P r e p a r a tio n Dissolve 0.500 g in carbon dioxide-free water R and dilute to
Benzylpenicillin Injection 25.0 m L with the same solvent.
P h E tr __________________________________ A b so rb a n c e (2.2.25)
Dissolve 90.0 m g in water R and dilute to 50.0 m L with the
D E F IN IT IO N
same solvent. M easure the absorbance o f the solution at
Sodium (2S,5R,6R)-3,3-dimethyl-7-oxo-6 -
325 nm , at 280 nm and at the absorption maximum at
[(phenylacetyl)amino]-4-thia-l-azabicyclo[3.2.0]heptane-2- 264 nm , diluting the solution, if necessary, for the
carboxylate.
m easurem ent at 264 nm. T h e absorbances at 325 nm and
Substance produced by the growth o f certain strains of 280 nm are not greater than 0.10 and the absorbance at the
PenicHtium notation or related organisms, or obtained by any absorption maximum at 264 nm is 0.80 to 0.88, calculated
other means. on the basis o f the undiluted (1.80 g/L) solution. Verify the
C o n te n t resolution o f the apparatus (2.2.25); the ratio of the
96.0 per cen t to 102.0 per cent (dried substance). absorbances is at least 1.7.
CHARACTERS R e la te d su b sta n c e s
A p p e a ra n c e Liquid chromatography (2.2.29). Prepare the solutions
W hite or alm ost white, crystalline powder. immediately before use.
2016 Benzylpenicillin Sodium 1-273
Test solution (a) Dissolve 50.0 mg o f the substance to be B a c te ria l en d o to x in s (2.6.14, Method E)
examined in water R and dilute to 50.0 m L with the same Less than 0.16 IU/m g, if intended for use in the manufacture
solvent. o f parenteral preparations without a further appropriate
Test solution (b) Dissolve 80.0 mg of die substance to be procedure for the removal of bacterial endotoxins.
examined in water R and dilute to 20.0 m L with the same ASSAY
solvent. Liquid chromatography (2.2.29) as described in the test for
Reference solution (a) Dissolve 50.0 m g of benzylpema&n related substances with the following modifications.
sodium CRS in water R and dilute to 50.0 m L with the same Mobile phase Initial composition o f the mixture of mobile
solvent. phases A and B, adjusted where applicable.
Reference solution (b) Dissolve 10 mg o f benzylpemdOin Injection T e st solution (a) and reference solution (a).
sodium CRS and 10 mg o f phenylacetic add R (impurity B) in
Calculate die percentage content o f C i 6H 17N 2N a 0 4S from
water R, then dilute to 50 m L with the same solvent.
die declared content o f benzylpemdSBn sodium CRS.
Reference solution (c) Dilute 4.0 m L o f reference solution (a)
to 100.0 m L with water R STO R A G E
Column: In an airtight container. If the substance is sterile, store in a
— size: I = 0.25 m , 0 = 4.6 mm; sterile, airtight, tamper-proof container.
— stationary phase: octadecylsifyl silica gelfor chromatography R IM P U R IT IE S
(5 Jim).
Mobile phase:
— mobile phase A: mix 10 volumes o f a 68 g/L solution of
potassium dihydrogen phosphate R adjusted to p H 3.5 with h 2n--
a 500 g/L solution of dilute phosphoric add R, 30 volumes
of methanol R and 60 volumes o f water R;
— mobile phase B: mix 10 volumes o f a 68 g/L solution of
A. (2S,5Æ,6i?)-6-amino-3,3-dimethyl-7-oxo-4-thia-l-
potassium dihydrogen phosphate R adjusted to p H 3.5 with azabicydo [3.2.0]heptane- 2-carboxylic ad d
a 500 g/L solution of dilute phosphoric add R, 40 volumes (6-aminopenidllanic add),
of water R and 50 volumes o f methanol R;
H C02H TESTS
HN'^X.CHa Related substances
H U / \ ru and epimerat C*
Determine the absorbance (2.2.25) o f test solutions (b)
ç r r h and (a) used in Identification, at 455 nm an d at 340 nm
respectively.
Absorbance ratio A 4 S5 / A ^ : minimum 1.5.
F. (2i?5,45)-2-[[(phenyIacetyl)amino]methyi]-5,5-
dimetiiylthiazoHdine-4-caiboxylic a d d (penifloic ad d s o f T h e thresholds indicated under Related substances
(Table 2034.-1) in the general monograph Substances for
benzylpenidllin).
pharmaceutical use (2034) do n o t apply.
PhEur
Heavy metals (2.4.8)
Maximum 10 ppm .
2.0 g complies with test D . Prepare the reference solution
using 2 m L o f lead standard solution (10 ppm Pb) R
★ ★
Betacarotene ★ ★ Loss on drying (2.2.32)
*
(Ph. Eur. monograph 1069) •k* M aximum 0.2 p er cent, determined on 1.000 g by drying
in vacuo over diphosphorus pentoxide R at 40 °C for 4 h.
Sulfated ash (2.4.14)
M aximum 0.2 p er cent, determined on 1.0 g, moistened with
a mixture o f 2 mT. o f dilute sulfuric add R and 5 m L o f
ethanol (96 per cent) R
ASSAY
Measure the absorbance <2.2.25) o f test solution (b) used in
Identification at the absorption maximum at 455 nm , using
cydohexane R as the compensation liquid.
Calculate the content o f C 40H 56 taking the specific
absorbance to be 2500.
STORAGE
In an airtight container, protected from light, at a
tem perature n o t exceeding 25 °C.
PhEur
C40H 56 536.9 7235-40-7
Content
96.0 per cent to 101.0 p e r cent (dried substance).
CHARACTERS
Appearance
Brown-red or brownish-red, crystalline powder.
Solubility
Practically insoluble in w ater, slightly soluble in cydohexane,
practically insoluble in anhydrous ethanol.
It is sensitive to air, heat a n d light) especially in solution.
Cany out all operations as rapidly as possible avoiding exposure to
actinic light; use freshly prepared solutions.
IDENTIFICATION
1135 7585-39-9
Ultraviolet and visible absoiption spectrophotometry (2.2.25).
Test solution (a) Dissolve 50.0 m g in 10 m L o f chloroform R Action and use
and dilute immediately to 100.0 m L with cydohexane R. Carrier molecule for drug delivery systems.
D ilute 5.0 m L o f this solution to 100.0 m L with
cydohexane R P h B r.
R e la te d su b sta n c e s
Carrier gas helium for chromatography R.
Liquid chrom atography (2.2.29). Static head-space conditions which may be used:
— equilibration temperature: 45 °C;
Test solution (a) Dissolve 0.25 g of the substance to be
— equilibration timer. 2 h.
examined in water R with heating, cool and dilute to
25.0 m L with the same solvent. Temperature:
— column: 50 °C;
Test solution (b) Dilute 5.0 m L of test solution (a) to
— infection port. 140 °C;
50.0 m L with water R.
— detector. 280 °C.
Reference solution (a) Dissolve 25.0 mg o f alfadex CRS
Detection Flame ionisation.
(impurity A), 25.0 mg o f gammacyclodextrin CRS
(impurity B) and 50.0 mg of betadex CRS in water R, then Irtjection 200 |jL of the head space, at least 3 times.
dilute to 50.0 m L with the same solvent. Retention time Toluene = about 10 min.
Reference solution (b) Dilute 5.0 m L o f reference solution (a)
to 50.0 m L with water R.
1-276 Betahistine Dihydrochloride 2016
System suitability:
— resolution: m in im u m 1.1 between the peaks due to
trichloroethylene and toluene; minim um 1.1 between the
peaks due to toluene and ethylene chloride;
— repeatability: maximum relative standard deviations of the
ratios o f the areas o f the peaks due to trichloroethylene
and toluene to th at of the peak due to ethylene chloride
o f 5 per cent.
Calculate the content o f trichloroethylene and of toluene
taking their relative densities to be 1.46 and 0.87,
respectively.
Limits:
— trichloroethylene: m axim um 10 ppm;
— toluene, maximum 10 ppm .
H eav y m e ta ls (2.4.8)
M axim um 10 ppm. B. cyclooctakis-(l -+4)-(a-D-glucopyranosyl)
1.0 g complies with test C. Prepare the reference solution (cyclomaltooctaose or y-cyclodextrin).
using 1 m L o f lead standard solution (10 ppm Pb) R.
L oss o n d ry in g (2.2.32)
M axim um 16.0 per cent, determ ined on 1.000 g by drying in
an oven at 120 °C for 2 h. ★* ★
★ ★
S u lfa te d a s h (2.4.14) Betahistine Dihydrochloride ★ ★
M axim um 0.1 per cent, determ ined on 1.0 g. * * * * *
(Ph. Eur. monograph 1665)
A SSA Y
Liquid chromatography (2.2.29) as described in the test for R
related substances w ith the following modifications. f CH3 ,2HCI
Injection T est solution (b) and reference solutions (a) and (c).
System suitability: reference solution (a):
— repeatability: maximum relative standard deviation of the C8Hi4C12N 2 209.1 5579-84-0
area of the peak due to betadex of 2.0 per cent.
A c tio n a n d u se
Calculate the percentage content of [C 6H 10O 5]7 from the
Histamine H ] receptor antagonist; antih istam in e.
assigned content of betadex CRS.
P r e p a r a tio n
STORAGE B etahistin e Dihydrochloride Tablets
In an airtight container.
PhEur___________________________________________________________
IM P U R IT IE S
Specified impurities A, B D E F IN IT IO N
N-M ethyl-2-(pyridin-2-yi)ethanamine dihydrochloride.
C o n te n t
99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
A p p e a ra n c e
W hite or slightly yellow powder, very hygroscopic.
S o lu b ility
Very soluble in water, soluble in ethanol (96 per cent),
practically insoluble in 2 -propanol.
ID E N T IF IC A T IO N
First identification B, D
Second identification A , C, D
A. M eltin g point (2.2.14): 150 °C to 154 °C.
A. cyclohexakis-(l-*4)-(a-D-glucopyranosyl) (alfadex or
B. Infrared absorption spectrophotom etry (2.2.24).
cyclomaltohexaose or a-cyclodextrin),
Comparison betahistine dihydrochloride CRS.
C. Thin-layer chrom atography (2.2.27).
Test solution Dissolve 10 m g of the substance to be examined
in 2 m L o f ethanol (96 per cent) R.
Reference solution Dissolve 10 m g o f betahistine
dihydrochloride CRS in 2 m L o f ethanol (96 per cent) R.
Plate TLC silica gel GF2s\ plate R.
Mobile phase concentrated ammonia R, ethyl acetate R,
methanol R (0.75:15:30 VIVIV).
Application 2 |iL.
2016 Betahistine Mesilate 1-277
Development Over 2/3 of the plate. — total: not more than 0.5 times the area of the principal
Drying At 110 °C for 10 min. peak in the chromatogram obtained with reference
solution (b) (0.5 per cent);
Detection Examine in ultraviolet light at 254 nm.
— disregard limit. 0.25 times the area o f the principal peak in
Results T he principal spot in the chromatogram obtained with the chromatogram obtained with reference solution (c)
the test solution is similar in position and size to the prindpal (0.05 p er cent).
spot in the chromatogram obtained with the reference
solution. L oss o n d ry in g (2.2.32)
M aximum 1.0 per cent, determined on 1.000 g by drying in
D . It gives reaction (a) o f chlorides (2.3.1).
an oven at 105 °C.
TESTS S u lfa te d a sh (2.4.14)
Solution S Maximum 0.1 p er cent, determined on 1.0 g.
Dissolve 5.0 g in carbon dioxide-free water R, and dilute to
50 m L with the same solvent. ASSAY
Dissolve 80.0 m g in 50 m L of ethanol (96 per cent) R. T itrate
Appearance o f solution
with 0.1 M sodium hydroxide, determ in in g the end-point
Solution S is clear (2.2.1) and not more intensely coloured
potentiometrically (2.2.20). Read the volume added to reach
than reference solution B 8 (2.2.2, Method II).
the second point of inflexion.
pH (2.2.3) 1 m L of 0.1 M sodium hydroxide is equivalent to 10.46 m g of
2.0 to 3.0 for solution S. C 8H 14C12N 2.
Related substances
STORA GE
Liquid chromatography (2.2.29).
In an airtight container.
Test solution Dissolve 25 mg o f die substance to be examined
in the mobile phase and dilute to 25.0 m L with the mobile IM P U R IT IE S
phase. Specified impurities A, B, C.
Reference solution (a) Dissolve 10 m g of betahistine
dihydrochloride CRS and 10 mg of 2-virtylpyridine R in the
mobile phase and dilute to 50.0 m L with the mobile phase. C T "
Dilute 2.0 m L o f the solution to 50.0 m L with the mobile
phase. A. 2-ethenylpyridine (2-vinylpyridine),
Reference solution (b) Dilute 1.0 m L of the test solution to
100.0 m L with the mobile phase.
Reference solution (c) Dilute 2.0 m L of reference solution (b)
to 10.0 m L with the mobile phase. C P '* '
Column:
— sizer. I = 0.15 m , 0 = 3.0 mm; B. 2-(pyridin-2-yl)ethanol,
— stationary phase: base-deactivated end-capped octadecylsilyl
ch3
silica gel for chromatography R (5 Jim).
n.
Mobile phase Dissolve 2.0 g o f sodium dodecyl sulfate R in a
mixture of 15 m L of a 10 per cent V/V solution o f sulfuric O T '- '^ O
acid R, 35 m L o f a 17 g/L solution of tetrabtaylammomum
hydrogen sulfate R and 650 m L of water R; adjust to p H 3.3 C. N-methyl-2-(pyridin-2-yl)-N-[2-(pyridin-2-
using dilute sodium hydroxide solution R and mix w ith 300 m L yl)ethyl] ethanamine.
of acetonitrile R.
PhEur
Flow rate 1 mlVmin.
Detection Spectrophotom eter at 260 nm.
Irtjection 20 (iL.
Run time 4 times the retention time of betahistine. ★ ★
Betahistine Mesiiate ★ ★
Relative retention W ith reference to betahistine (retention
(Ph. Eur. monograph 1071) * * * * *
time = about 7 m in): impurity B = about 0.2;
impurity A = about 0.3; impurity C = about 3.
System suitability: reference solution (a):
— resolution: minim um 3.5 between the peaks due to CH3 , 2 H3C -S O 3H
2-vinylpyridine and betahistine.
Limits:
— correction factor, for the calculation of content, multiply the C 10H20N2O6S2 328.4 54856-23-4
peak area o f impurity B by 0.4;
— impurities A , B, C: for each impurity, n o t more th an the A ctio n a n d u se
area o f the principal peak in the chromatogram obtained Histamine H I receptor antagonist; antihistamine.
with reference solution (c) (0.2 per cent);
PhEur_________________________________________________________
— unspecified impurities: for each impurity, not m ore than
0.5 times o f the area o f the principal peak in the D E F IN IT IO N
chromatogram obtained with reference solution (c) N-Methyl-2-(pyridin-2-yl)ethanamine bis(methanesulfonate).
(0.10 p er cent); C o n te n t
98.0 per cent to 101.0 per cent (anhydrous substance).
1-278 Betahistine Mesilate 2016
T itrate with 0.1 M perchloric acid, determining the end-point chloride R, evaporate to dryness on a water-bath and record
potentiometrically (2.2.20). new spectra using the residues.
1 m L of 0.1 M perchloric add is equivalent to 16.42 mg C. Thin-layer chromatography (2.2.27).
of C 10H 20N 2O 6S 2. Solvent mixture methanol R, methylene chloride R (1:9 V/V).
STORA GE Solvent mixture
In an airtight container. Test solution Dissolve 10 mg of the substance to be examined
IM P U R IT IE S in the solvent mixture and dilute to 10 m L with the solvent
Specified impurities A mixture.
Reference solution (a) Dissolve 20 m g of betamethasone CRS in
the solvent mixture and dilute to 20 m L with the solvent
mixture.
C T "
Reference solution (b) Dissolve 10 m g of dexamethasone CRS in
reference solution (a) and dilute to 10 m L with reference
A. 2-ethenylpyridine (2-vinylpyridine).
solution (a).
PhEur
Plate TLC silica gel F 254 plate R.
Mobile phase butanol R saturated with water R, toluene R,
ether R (5:10:85 VIVIV).
★ ★ Application 5 |iL.
Betamethasone ★ ★
Development Over a path of 15 cm.
*****
(Ph. Eur. monograph 0312) Drying In air.
Detection A Examine in ultraviolet light at 254 nm.
Results A T h e principal spot in the chromatogram obtained
with the test solution is similar in position and size to the
principal spot in the chromatogram obtained with reference
solution (a).
Detection B Spray with alcoholic solution of sulfuric acid R. H eat
at 120 °C for 10 m in or until the spots appear. Allow to
C22H 29FO 5 392.5 378-44-9 cool. Examine in daylight and in ultraviolet light at 365 nm .
Results T h e principal spot in the chromatogram obtained w ith
A ctio n a n d u se the test solution is similar in position, colour in daylight,
Glucocorticoid. fluorescence in ultraviolet light at 365 nm and size to the
P re p a ra tio n principal spot in the chromatogram obtained with reference
Betamethasone T ablets solution (a).
System suitability: reference solution (b):
PhEir ___________________ — the chromatogram shows 2 spots which may, however,
D E F IN IT IO N not be completely separated.
9-Fluoro-l 1P, 17 j2 1-trihydroxy-16 ^-methylpr egna-1,4- D . Mix about 5 m g with 45 mg of heavy magnesium oxide R
diene-3,20-dione. and ignite in a crucible until an almost white residue is
C o n te n t obtained (usually less than 5 min). Allow to cool, add 1 m L
97.0 per cent to 103.0 per cent (dried substance). o f water R, 0.05 m L of phenolphthalein solution R1 and about
1 m L of dilute hydrochloric acid R to render the solution
CHARACTERS colourless. Filter. Add 1.0 m L of the filtrate to a freshly
A p p earan ce prepared mixture o f 0.1 m L of alizarin S solution R and
White or almost white, crystalline powder. 0.1 m L o f zirconyl nitrate solution R. Mix, allow to stand for
Solubility 5 min and compare the colour of the solution with that o f a
Practically insoluble in water, sparingly soluble in anhydrous blank prepared in the same manner. T he test solution is
ethanol, very slightly soluble in methylene chloride. yellow and the blank is red.
ID E N T IF IC A T IO N E. Add about 2 m g to 2 m L of sidfuric acid R and shake to
First identification B, C. dissolve. W ithin 5 min, a deep reddish-brown colour
develops. Add this solution to 10 m L of water R and mix.
Second identification A , C, D, E.
T h e colour is discharged and a clear solution remains.
A. Dissolve 10.0 m g in anhydrous ethanol R and dilute to
100.0 m L with the same solvent. Place 2.0 m L of this TESTS
solution in a stoppered tube, add 10.0 m L o f S p ecific o p tic a l ro ta tio n (2.2.7)
phertylhydrazine-sidfuric acid solution R, mix and heat in a + 118 to + 126 (dried substance).
water-bath at 60 °C for 20 min. Cool immediately. Dissolve 0.125 g in methanol R and dilute to 25.0 m L with
The absorbance (2.2.25) measured at 419 n m is n o t greater the same solvent.
than 0 . 10. R e la te d su b sta n c e s
B. Infrared absorption spectrophotometry (2.2.24). Liquid chromatography (2.2.29).
Comparison betamethasone CRS. Test solution Dissolve 25.0 mg o f the substance to be
If the spectra obtained in the solid state show differences, examined in a mixture of equal volumes o f acetonitrHe R and
dissolve the substance to be examined and the reference methanol R and dilute to 10.0 m L with the same mixture o f
substance separately in the minimum volume of methylene solvents.
1-280 Betamethasone 2016
Reference solution (a) Dissolve 2 m g o f betamethasone CRS and Calculate the content of C 22H 29F O 5 taking the specific
2 m g of methylprednisolone CRS in mobile phase A, then absorbance to be 395.
dilute to 100.0 m L with mobile phase A. STO RA G E
Reference solution (b) Dilute 1.0 m L of the test solution to Protected from light.
100.0 m L with mobile phase A.
IM P U R IT IE S
Column'.
Specified impurities A, B, C, D , E, F, G , H , I, J
— size. I = 0.25 m , 0 = 4.6 mm;
— stationary phase, octadecylsilyl silica gel for chromatography R
(5 Jim);
— temperature: 45 °C.
Mobile phase".
— mobile phase A: in a 1000 m L volumetric flask mix
250 m l. o f acetomtrUe R w ith 700 m L of water R and
allow to equilibrate; dilute to 1000 m L with water R and
mix again; A. 9 -flu o ro -lip , 17,21 -trihydroxy-16a-m ethylpregna-1,4-
— mobile phase B: acetomtrUe R; diene-3,20-dione (dexamethasone),
4 0-41 0 * 100 10 0 * 0
4 1 -4 6 100 0
B. 21-chloro-9-fluoro-l ip,17-dihydroxy-16P-methylpregna-
l,4-diene-3,20-dione,
Flow rate 2.5 mlVmin.
Detection Spectrophotom eter at 254 nm .
Equilibration W ith mobile phase B for at least 30 min and
then with mobile phase A for 5 min. F or subsequent
chromatograms, use the conditions described from 40 min to
46 min.
Injection 20 pL; inject the mixture of equal volumes o f
acetomtrUe R and methanol R asa, blank. C. 17,21-dihydroxy-16P-methylpregna-l,4,9(l l)-triene-3,20-
Retention time M ethylprednisolone = about 11.5 min; dione,
betam ethasone = about 12.5 min.
System suitability: reference solution (a):
— resolution: minimum 1.5 between the peaks due to
methylprednisolone and betamethasone; if necessary,
adjust the concentration o f acetonitrfle in mobile phase A.
Limits:
— impurities A , B, C, D, E, F, G, H, 1, J: for each impurity,
no t more than the area o f the principal peak in the
chrom atogram obtained w ith reference solution (b) D . 9 -flu o ro -lip , 17-dihydroxy-16P-methyl-3,20-dioxopregna-
( 1.0 per cent), and not m ore than 1 such peak has an l,4-dien-21-yl ethoxycarboxylate,
area greater than 0.5 times the area of the principal peak
in the chrom atogram obtained with reference solution (b)
(0.5 per cent);
— total: not more than twice the area of the principal peak in
~ the chrom atogram obtained with reference solution (b)
( 2.0 per cent);
— disregard limit. 0.05 times the area of the principal peak in
the chrom atogram obtained with reference solution (b)
(0.05 per cent). E. 9,11 P-epoxy-17,21 -dihydroxy-16P-methyl-9 P-pregna-1,4-
L oss o n d ry in g (2.2.32) diene-3,20-dione,
M axim um 0.5 per cent, determ ined on 0.500 g by drying in
an oven at 105 °C.
A SSA Y
Dissolve 0.100 g in ethanol (96 per cent) R and dilute to
100.0 m L w ith the same solvent. Dilute 2.0 m L of this
solution to 100.0 m L with ethanol (96 per cent) R. M easure
the absorbance (2.2.25) at the absorption maximum at
238.5 nm. F . 17,21-dihydroxy-16P-methylpregna-l,4,l l-triene-3,20-
dione,
2016 Betamethasone Acetate 1-281
CHARACTERS
A p p e a ra n c e
W hite or almost white, crystalline powder.
S olu b ility
Practically insoluble in water, freely soluble in acetone,
soluble in ethanol (96 per cent) and in methylene chloride.
It shows polymorphism (5.9).
G . 1 la,17,21-trihydroxy-16ß-methylpregna-l,4-diene-3,20-
dione, ID E N T IF IC A T IO N
First identification B, C
Second identification A , C, D, E, F
A. Dissolve 10.0 m g in anhydrous ethanol R and dilute to
100.0 m L with the same solvent. Place 2.0 m L o f this
solution in a groimd-gjass-stoppered tube, add 10.0 m L o f
phenyJhydrazme-sulfuric acid solution R, m ix and heat in a
water-bath at 60 °C for 20 m in. Cool im m ediately.
T he absorbance (2.2.25) measured at 419 nm is not greater
H . 14-fluoro-l lß,17,21-trihydroxy-16ß-methyl-8a,9ß,14ß- than 0 . 10.
pregna-1 ,4-diene-3,20-dione, B. Infrared absorption spectrophotometry (2.2.24).
Comparison betamethasone acetate CRS.
If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and the reference
substance separately in the m inim um volume of methanol R,
evaporate to dryness on a water-bath and record new spectra
using the residues.
C. Thin-layer chromatography (2.2.27).
Solvent mixture methanol R, methylene chloride R (1:9 VIV).
I. 8-fluoro-llß,17,21-trihydroxy-16ß-methyl-8a,9ß-pregna-
l 34-diene-3,20-dione, Test solution Dissolve 10 mg of the substance to be exam ined
in the solvent mixture and dilute to 10 m L with the solvent
mixture.
Reference solution (a) Dissolve 20 mg of betamethasone
acetate CRS in the solvent mixture and dilute to 20 m L with
the solvent mixture.
Reference solution (b) Dissolve 10 mg of prednisolone
acetate CRS in reference solution (a) and dilute to 10 m L
with reference solution (a).
J. 17,21 -dihydroxy-16 (J-methylpregna-134-diene-3 ,2 0-dione. Plate TLC silica gel F 254 plate R.
__________________________________________________________ PhEtr Mobile phase Add a mixture of 1.2 volumes of water R and
8 volumes of methanol R to a mixture of 15 volumes of
ether R and 77 volumes o f methylene chloride R.
Application 5 pL.
Development Over a path o f 15 cm.
Betamethasone Acetate ******
Drying In. air.
(Ph Eur monograph 0975) * Detection A Examine in ultraviolet light at 254 nm.
Results A T h e principal spot in the chromatogram obtained
with the test solution is similar in position and size to the
principal spot in the chromatogram obtained with reference
solution (a).
Detection B Spray with alcoholic solution of sulfuric acid R. H eat
at 120 °C for 10 m in or until the spots appear. Allow to
cool. Examine in daylight and in ultraviolet light at 365 n m .
Results B T h e principal spot in the chromatogram obtained
C24H31F 0 6 434.5 987-24-6 with the test solution is similar in position, colour in daylight,
fluorescence in ultraviolet light at 365 nm and size to the
A ctio n a n d u se
principal spot in the chromatogram obtained with reference
Glucocorticoid.
solution (a).
PhEur__________________________________________________________ System suitability: reference solution (b):
— the chromatogram shows 2 clearly separated spots.
D E F IN IT IO N
D . Add about 2 m g to 2 m L o f sulfuric acid R and shake to
9-Fhioro-11(3,17-dihydroxy-16|}-inethyl-3,20-dioxopregna-
l,4-diene-21-yl acetate. dissolve. W ithin 5 min, a deep reddish-brown colour
develops. Add this solution to 10 m L of water R and mix.
C o n te n t T h e colour is discharged and a clear solution rem ain s.
97.0 per cent to 103.0 per cent (anhydrous substance).
1-282 Betamethasone Acetate 2016
o
*****
Betamethasone Sodium ★ ★
o< \ / o CH3
Phosphate *****
(Ph. Eur. monograph 0810)
r H V -C H 3
° ,ONa
0 " p\
"OH oNa
D. 2 l-(acetyloxy)-9-fluoro-l 1ß-hydroxy-16ß-methyl-3520-
dioxopregna-1,4-dien-17-yi propanoate (betamethasone
21-acetate 17-propionate),
Mobile phase glacial acetic add R, water R, butanol R Test solution Dissolve 62.5 m g o f the substance to be
(20:20:60 VIVIV). examined in the mobile phase and dilute to 25.0 m L with
Application 5 pL. the mobile phase.
Development Over a path of 15 cm. Reference solution (a) Dissolve 25 m g of betamethasone sodium
Drying In air. phosphate CRS and 25 m g o f dexamethasone sodium
phosphate CRS in the mobile phase and dilute to 25.0 m L
Detection A Examine in ultraviolet light at 254 nm. with the mobile phase. D ilute 1.0 m L o f this solution to
Results A T he principal spot in the chromatogram obtained 25.0 m L with the mobile phase.
with the test solution is similar in position and size to the Reference solution (b) Dilute 1.0 m L of the test solution to
principal spot in the chromatogram obtained with reference 50.0 m L with the mobile phase.
solution (a).
Column:
Detection B Spray with alcoholic solution of sulfuric add R. H eat — size: I = 0.25 m , 0 = 4.6 mm;
at 120 °C for 10 min or until the spots appear. Allow to — stationary phase: octadecylsifyl silica gel for chromatography R
cool. Examine in daylight and in ultraviolet light at 365 nm. (5 pm).
Results B T h e principal spot in the chromatogram obtained Mobile phase In a 250 m L conical flask, weigh 1.360 g of
with the test solution is similar in position, colour in daylight, potassium dihydrogen phosphate R and 0.600 g o f hexylamine R,
fluorescence in ultraviolet light at 365 nm and size to the mix and allow to stand for 10 min and then dissolve in
principal spot in the chromatogram obtained with reference 185 m L o f water R\ add 65 m L of acetomtrUe R, mix and
solution (a). filter (0.45 pm).
System suitability: reference solution (b): Flow rate 1 mL/min.
— the chromatogram shows 2 spots which may, however,
Detection Spectrophotom eter at 254 nm .
not be completely separated.
Equilibration W ith the mobile phase for about 45 min.
D. Add about 2 mg to 2 m L o f sulfuric acid R and shake to
dissolve. W ithin 5 m in, an intense reddish-brown colour Injection 20 pL.
develops. Add the solution to 10 m L of water R and mix. Run time Twice the retention time o f betamethasone sodium
T he colour is discharged and a clear solution remains. phosphate.
E. Mix about 5 mg with 45 mg of heavy magnesium oxide R Retention time Betam ethasone sodium phosphate = about
and ignite in a crucible until an almost white residue is 14 min; dexamethasone sodium phosphate = about
obtained (usually less than 5 min). Allow to cool, add 1 m L 15.5 min.
o f water R, 0.05 m L o f phenolphthakin solution R1 and about System suitability, reference solution (a):
1 m L of dilute hydrochloric add R to render the solution — resolution: minim um 2.0 between the peaks due to
colourless. Filter. Add 1.0 m L of die filtrate to a freshly betam ethasone sodium phosphate and dexamethasone
prepared mixture o f 0.1 m L of alizarin S solution R and sodium phosphate; if necessary, increase the
0.1 m L o f zvrconyl nitrate solution R. Mix, allow to stand for concentration o f acetonitrile or increase the concentration
5 min and compare the colour of the solution with th at o f a of water in die mobile phase.
blank prepared in the same manner. T h e test solution is Limits:
yellow and the blank is red. — any impurity, for each impurity, no t more than the area of
F. T o about 40 m g add 2 m L of sulfuric acid R and heat the principal peak in the chromatogram obtained with
gently until white fumes are evolved. Add nitric add R reference solution (b) (2 per cent), and not more than
dropwise, continue the heating until the solution is almost 1 such peak has an area greater than 0.5 times the area of
colourless and cool. A dd 2 m L o f water R, heat until white the principal peak in the chromatogram obtained with
fumes are again evolved, cool, add 10 m L of water R and reference solution (b) (1 per cent);
neutralise to red litmus paper R with dilute ammonia R l. — total: n o t more than 1.5 times the area of the principal
T he solution gives reaction (a) of sodium (2.3.1) and peak in the chrom atogram obtained with reference
reaction (b) o f phosphates (2.3.1). solution (b) (3 per cent);
TESTS — disregard limit: 0.025 times the area o f the principal peak
in the chromatogram obtained with reference solution (b)
S o lu tio n S
(0.05 per cent).
Dissolve 1.0 g in carbon dioxide-free water R and dilute to
20 m L with the same solvent. In o rg a n ic p h o sp h a te
M aximum 1 per cent.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2 . 2 . 1 ) and n o t m ore intensely coloured Dissolve 50 m g in water R and dilute to 100 m L with the
than reference solution B 7 (2.2.2, Method II). same solvent. T o 10 m L o f this solution add 5 m L of
molybdovanadic reagent R, mix and allow to stand for 5 min.
p H (2.2.3)
Any yellow colour in the solution is not m ore intense than
7.5 to 9.0.
that in a standard prepared at the same time and in the same
Dilute 1 m L o f solution S to 5 m L with carbon dioxide-free maimer using 10 m L of phosphate standard solution (5 ppm
water R. PO4 ) R.
Specific o p tic a l r o ta tio n (2.2.7) W a te r (2.5.12)
+ 98 to + 104 (anhydrous substance). M aximum 8.0 per cent, determined on 0.200 g.
Dissolve 0.250 g in water R and dilute to 25.0 m L with the
ASSAY
same solvent.
Dissolve 0.100 g in water R and dilute to 100.0 m L with the
R elated su b s ta n c e s same solvent. D ilute 5.0 m L o f this solution to 250.0 m L
Liquid chrom atography (2.2.29). with water R. M easure the absorbance (2.2.25) at the
absorption maximum at 241 nm.
2016 Betamethasone Valerate 1-287
A c tio n a n d u se
Beta-adrenoceptor antagonist.
Preparations
Betaxolol Eye Drops, Solution
A. R l = R3 = R5 = R 6 = H , R2 = F , R4 = C H 3: 9-fluoro-
Betaxolol Eye Drops, Suspension
1 l ß ,17,21 -trihydroxy-16 ß-methylpregna-1,4-diene-3,20-dione
(betamethasone), PhEur-----------------------------------------------------------------------------------------
C. R l = R 4 = R 6 = H , R2 = F, R3 = C H 3, R5 = CO- D E F IN IT IO N
[C H 2] 3-C H 3: 9-fluoro-l 1ß,21-dihydroxy-l 6a-methyl-3,20-
(2RS) -1 -[4- [2-(Cyclopropylmethoxy) ethyl] phenoxy] -3-
dioxopregnal,4-dien-17-yl pentanoate (dexamethasone
[(l-m ethylediyl)amino]propan- 2-ol hydrochloride.
17-valerate),
C o n te n t'
E. R l = R3 = R5 = H , R2 = F , R4 = C H 3, R 6 = C O -
98.5 per cent to 101.5 per cent (dried substance).
[ C H J 3-C H 3: 9-fluoro-l 1ß, 17-dihydroxy-l6ß-methyl-3,20-
dioxopregna 1,4-dien-21 -yl pentanoate (betamethasone CHARACTERS
21-valerate), A p p e a ra n c e
G. R l = Br, R2 = F, R3 = R 6 = H , R4 = C H 3, R5 = W hite or almost white, crystalline powder.
C O- [CH 2] 3-C H 3: 6a-brom o-9-fluoro-l 1ß,21-dihydroxy- S o lu b ility
16ßmethyl-3,20-dioxopregna-l ,4-dien-17-yl pentanoate Very soluble in water, freely soluble in ethanol (96 p er cent),
( 6a-brom o-betam ethasone valerate), soluble in methylene chloride.
H . R l = R3 = R 6 = H , R2 = Cl, R4 = C H 3, R5 = C O - ID E N T IF IC A T IO N
[C H J 3-C H 3: 9-chloro-11 ß,2 1-dihydroxy-16 ß-methyl-3,20-
First identification B, D.
dioxopregna 1,4-dien-17-yl pentanoate (beclomethasone
17-valerate), Second identification A , C, D.
A. M elting point (2.2.14): 113 °C to 117 °C.
I. R l = R3 = R4 = R 6 = H , R2 = F , R5 = C O - f C H ^ -
C H 3: 9-fluoro-l lß ,21-dihydroxy-3,20-dioxopregna-1,4-dien- B. Infrared absorption spectrophotom etry (2.2.24).
17-yi pentanoate (9-fluoro-prednisolone 17-valerate), Comparison betaxold hydrochloride CRS.
2016 Betaxolol Hydrochloride 1-289
C . Thin-layer chromatography (2.2.27). — stationary phase: octylsUyl silica gel for chromatography R
Test solution Dissolve 10 mg of the substance to be examined (5 nm).
in 1 m L o f methanol R. Mobile phase M ix 175 m L of acetonitrile R and 175 m L of
Reference solution (a) Dissolve 20 mg of betaxolol methanol R and dilute to 1 L with a 3.4 g/L solution of
hydrochloride CRS in 2 m L of methanol R. potassium dihydrogen phosphate R, previously adjusted to
p H 3.0 with phosphoric add R.
Reference solution (b) Dissolve 10 m g of oxprenolol
hydrochloride CRS in 1 m L of reference solution (a). Flow rate 1.5 mL/min.
Plate TLC octadecylsHyl silica gel F 254 pl&te R- Detection Spectrophotometer at 273 nm .
Mobile phase perchloric add R, methanol R, water R Injection 20 nL o f the test solution and reference
(0.5:50:50 VIVIV). solutions (a), (b) and (d).
Application 2 (xL. Run time 4.5 the retention time of betaxolol.
Development Over a path of 10 cm. Identification of impurities Use the chromatogram obtained
with reference solution (a) to identify the peak due to
Drying In air.
impurity A; use the chromatogram supplied with betaxolol for
System suitability-, reference solution (b): peak identification CRS and the chromatogram obtained with
— the chromatogram shows 2 clearly separated spots.
reference solution (d) to identify the peaks due to
Detection A Examine in ultraviolet light at 254 nm . impurities B, C, D and E.
Results A T h e principal spot in the chromatogram obtained Relative retention W ith reference to betaxolol (retention
with the test solution is similar in position and size to the time = about 8 min): impurity B = about 0.3;
principal spot in the chromatogram obtained with reference impurity A = about 0.8; impurity D = about 1.5;
solution (a). impurity E = about 2.2; impurity C = about 4.1.
Detection B Spray with a 50 g/L solution o f vanillin R in a System suitability: reference solution (a) :
mixture o f 5 volumes of sulfuric add R, 10 volumes of glacial — resolution: m in im u m 2.0 between the peaks due to
acetic acid R and 85 volumes of methanol R, heat at impurity A and betaxolol.
100-105 °C until the colour o f the spots reaches maximum Limits:
intensity (10-15 min), and examine in daylight. — impurities A , B, C, D, E: for each impurity, not more than
Results B T he principal spot in the chromatogram obtained 0.3 times the area o f the prindpal peak in the
w ith the test solution is similar in position, colour and size to chromatogram obtained with reference solution (b)
the prindpal spot in the chromatogram obtained with (0.3 per cent);
reference solution (a). — unspecified impurities: for each impurity, not more than
D . It gives reaction (a) of chlorides (2.3.1). 0.1 times the area o f the principal peak in the
chromatogram obtained with reference solution (b)
TESTS
(0.10 per cent);
Appearance o f solution — total: not more than the area of the prindpal peak in the
T h e solution is clear (2.2.1) and colourless (2.2.2,
chromatogram obtained with reference solution (b)
Method II). (1.0 per cent);
Dissolve 0.5 g in water R and dilute to 25 m L w ith the same — disregard limit. 0.05 times the area o f the prindpal peak in
solvent. the chromatogram obtained with reference solution (b)
Acidity or alkalinity (0.05 per cent).
Dissolve 0.20 g in carbon dioxide-free water R and dilute to H eav y m e ta ls (2.4.8)
20 m L with the same solvent. Add 0.2 m L of methyl red M aximum 10 ppm .
solution R and 0.2 m L of 0.01 M hydrochloric add. Dissolve 2.0 g in 20 m L of water R. 12 m L of the solution
T h e solution is red. Add 0.4 m L of 0.01 M sodium hydroxide.
complies with test A. Prepare the reference solution using
T h e solution is yellow.
10 m L o f lead standard solution (1 ppm Pb) R.
Related substances L oss o n d ry in g (2.2.32)
L iquid chromatography (2.2.29). Prepare reference solutions (c)
M aximum 0.5 per cent, determined on 1.000 g by drying in
and (d) immediately before use. an oven at 105 °C.
Test solution Dissolve 10 mg of the substance to be examined
S u lfa te d a sh (2.4.14)
in the mobile phase and dilute to 5.0 m L with the mobile
M aximum 0.1 per cent, determined on 1.0 g.
phase.
Reference solution (a) Dissolve 8 mg of the substance to be ASSAY
examined and 4 m g o f betaxold impurity A CRS in 20.0 m L Dissolve 0.300 g in a mixture of 10.0 m L of 0.01 M
of the mobile phase. hydrochloric add and 50 m L of ethanol (96 per cent) R. Carry
out a potentiometric titration (2.2.20), using 0.1 M sodium
Reference solution (b) Dilute 1.0 m L of the test solution to
hydroxide. Read the volume added between the 2 points of
100.0 m L with the mobile phase.
inflexion.
Reference solution (c) Dissolve 2 m g o f betaxolol
1 m L of 0.1 M sodium hydroxide is equivalent to 34.39 m g
impurity C CRS in 50 m L of the mobile phase. D ilute 5 m L
of C 18H 30CINO 3.
o f the solution to 20 m L with the mobile phase.
Reference solution (d) Dissolve 10 m g o f betaxolol for peak STORA GE
identification CRS (containing impurities B, D and E) in Protected from light.
5 m L of reference solution (c). IM P U R IT IE S
Column'. Specified impurities A, B, C, D, E
— sizer. I = 0.25 m , 0 = 4 mm; •
1-290 Bezafibrate 2016
H OH
H CHARACTERS
N ^ CH3
T
^ „
and enantiomer Appearance
H3C CH3 W hite or almost white, crystalline powder.
Solubility
A. (2ftS)-l-(4-ethylphenoxy)-3- Practically insoluble in water, freely soluble in
[(1 -methylethyl)amino] propan-2-ol, dimethylformamide, sparingly soluble in acetone and in
ethanol (96 per cent). It dissolves in dilute solutions o f alkali
H OH hydroxides.
N ^/C H 3
I and enantiomer It shows polymorphism (5.9).
CH3 IDENTIFICATION
First identification A , B.
B. (2RS)-1- [4-(2-hydroxyethyI)phenoxy]-3- Second identification A, C.
[(1 -methylethyl) amino] propan-2-ol, A. Melting point (2.2.14): 181 °C to 185 °C.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison bezafibrate CRS.
and enantiomer
If the spectra obtained show differences, dissolve the
substance to be examined and the reference substance
separately in methanol R and evaporate to dryness. D ry the
C. (2.RS)-2-[[4-[2- residues in vacuo at 80 °C for 1 h and record new spectra
(cyclopropylmethoxy)ethyl] phenoxy] methyl] oxirane, using the residues.
C. Thin-layer chromatography (2.2.27).
OH Test solution Dissolve 10 mg o f the substance to be examined
in methanol R and dilute to 5 m L with the sam e solvent.
Reference solution Dissolve 10 mg o f bezafibrate CRS in
methanol R and dilute to 5 m L with the same solvent.
D . 4 -[2-(cyclopropylmethoxy)ethyl]phenol, Plate TLC silica gel F2sa R
Mobile phase glacial acetic acid R, methyl ethyl ketone R,
H OH xylene R (2.7:30:60 V/V/V).
Application 5 pL.
Development Over h alf of the plate.
and enantiomer Drying A t 120 °C for at least 15 min.
Detection Examine in ultraviolet light at 254 nm .
E. (2i?5)-l-[4-(2-butoxyethyl)phenoxy]-3- Results T he principal spot in the chrom atogram obtained with
[(1 -methylethyl) amino] propan-2-ol. the test solution is similar in position and size to the principal
PhEur spot in the chromatogram obtained with the reference
solution.
TESTS
Solution S
Bezafibrate ***** Dissolve 1.0 g in dimethylformamide R and dilute to 20 mL
★ ★
* ★ with the same solvent.
(Ph Eur monograph 1394) Appearance o f solution
Solution S is clear (2.2.1) and n o t m ore intensely coloured
O ^ C O jH
than reference solution BY5 (2.2.2, Method II).
H3C CH3 Related substances
Liquid chromatography (2.2.29).
a Test solution Dissolve 50.0 m g o f the substance to be
examined in the mobile phase and dilute to 100.0 m L with
C 19H 20CINO4 361.8 41859-67-0 the mobile phase.
Action and use Reference solution (a) Dilute 10.0 m L o f the test solution to
Fibrate; lipid-regulating drug. 100.0 m L with the mobile phase. D ilute 5.0 m L o f this
solution to 100.0 m L with the mobile phase.
Preparations
Reference solution (b) Dilute 5.0 m L o f reference solution (a)
Bezafibrate Tablets
to 50.0 m L with the mobile phase.
Prolonged-release Bezafibrate Tablets
Reference solution (c) T o 1 m L o f the test solution, add 1 m L
PhEur. o f 0.1 M hydrochloric add and evaporate to diyness on a hot
plate. Dissolve the residue in 20 m L o f the mobile phase.
DEFINITION
Column:
2-[4-[2-[(4-Chlorobenzoyl)amino]ethyl]phenoxy]-2-
— sizer. I = 0.125 m , 0 = 4 mm ;
methylpropanoic acid.
— stationary phase: octadecylsHyl silica gd for chromatography R
Content (5 pm)'.
98.0 per cent to 102.0 per cent (dried substance).
2016 Bicalutamide 1-291
O OHO CH,
^ 3andenan**0mer *'/ \f
sn A co 2h and enantiomer
. 8
C. (2iiS)-N-t4-cyano-3-(trifluoromethyl)phenyl]-3-
[(4-fluorophenyl)sulfonyI]-2-methylpropanainidej
M. (2RS) -3-[(4-fluorophenyl) sulfonyl] -2-hydroxy-2-
methylpropanoic add.
H2NV ^ s^ CF3
XX CN
PhEur
D. 4-amino-2-(txifluoromethyl)benzonitxile3 ***
★ ★
Bifonazole ★ ★
° “ v /^ H *****
(Ph. Ettr. monograph 1395)
and enantiomer
x r^ x c
and enantiomer
E. (2RS)-N- [4-cyano3-(trifluoromethyI)phenyl]-3-[(&S)-
(4-fluorophenyl)sulfinyl]-2-hydroxy-2-methylpropanamide,
°,
C22H 18N 2 310.4 60628-96-8
III I
I I a
nde
nan
ti
o
mer
fA ^ o v A cn Action and use
Antifungal.
F. (2SR)-N -[4-cyano-3-(trifluoromethyI)phenyl]-3 - [(RS)-
PhEir ____________
(4-fluorophenyl)sulfmyI]-2-hydroxy-2-methylpropanamide,
DEFINITION
1- [(&S)-(Biphenyl-4-yl) phenylmethyl] -1 //-imidazole.
nc
x> y w y -toc CN TESTS
Related substances
K. (22?,2 'S)-3,3 '-sulfonylbis [N-[4-cyano-3- Liquid chrom atography (2.2.29).
(trifluoromethyl)phenyI]-2-hydroxy-2-methylpropanamide], Buffer solution pH 32 M ix 2.0 m L o f phosphoric acid R with
980 m L o f water R, adjust to p H 3.2 (2.2.3) with
tnethylamine R and dilute to 1000.0 m L with water R.
1-294 Bifonazole 2016
12 - 30 10 90
NH
ID E N T IF IC A T IO N ASSAY
First identification A Suspend 0.200 g in 5 m L of dimethytformannde R. H eat until
the substance has dissolved completely. A dd 50 m L of
Second identification B, C
ethanol R and titrate with 0.1 M tetrabutylammonium
A. Examine by infrared absorption spectrophotometry
hydroxide, determining the end-point potentiometrically
( 2.2.24), comparing with the spectrum obtained with ( 2. 2. 20).
biomt CRS.
1 m L of 0.1 M tetrabutylammonium hydroxide is equivalent to
B. Examine the chromatograms obtained in the test for 24.43 mg o f CioH16N20 3S.
related substances (see Tests). The principal spot in the
chromatogram obtained with test solution (b) is similar in STORA GE
position and size to the principal spot in the chrom atogram Store protected from lig h t
obtained with reference solution (a). IM P U R IT IE S
C. Dissolve about 10 mg in 20 mL of water R with heating.
Allow to cool. Add 0.1 m L o f bromine water R. T he brom ine
water is decolourised.
*- = o = <
TESTS
S o lu tio n S
Dissolve 0.250 g in a 4 g/L solution o f sodium hydroxide R
and dilute to 25.0 m L with the same alkaline solution.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and colourless (2.2.2, Method IT). c o 2h
, cc>2h
and enantiomer Reference solution (a) Dissolve 25 m g of biperiden
H CH3 hydrochloride CRS in methanol R and dilute to 5 m L with the
same solvent.
D. 2-methyl-5-[(3a£,4S,6aR)-2-oxohexahydrothieno[3,4- Reference solution (b) Dissolve 5 m g o f biperiden
d\ imidazol-4-yl] pentanoic ad d , impurity A CRS in reference solution (a) and dilute to 2 m L
with the same solution.
Plate TLC silica gel F2 5 a plate R.
Mobile phase diethylamine R, methanol R, toluene R
(1:1:20 VfVfV).
Application 5 |iL
Development Over a path of 15 cm.
Drying In air.
c o 2h
Detection A Examine in ultraviolet light at 254 nm.
Results A T he prindpal spot in the chromatogram obtained
with the test solution is similar in position and size to the
prindpal spot in the chromatogram obtained with reference
solution (a).
E. 5- [(385,45,6aR)-3-benzyl-2-oxohexahydrothieno [3,4-
d\imidazol-4-yl]pentanoic a d d and 5-[(3a£,4S,6ai?)-l-benzyl- Detection B Spray with dilute potassium iodobismuthate
2-oxohexahydrothieno [334-d] imidazol-4-yl] pentanoic add.
solution R and then with sodium nitrite solution R and exam in e
in daylight.
___________________________________________________ ________ PhEur
Results B T he prindpal spot in the chromatogram obtained
with the test solution is similar in position, colour and size to
the prindpal spot in the chromatogram obtained with
reference solution (a).
***
Biperiden Hydrochloride ★ ★ System suitability, reference solution (b):
★ ★
— the chromatogram shows 2 clearly separated spots.
(Ph Eur monograph 1074) *****
C. T o about 20 m g add 5 m L o f phosphoric add R. A green
colour develops.
D . It gives reaction (a) of chlorides (2.3.1).
TESTS
and enantiomer , HCI S o lu tio n S
Dissolve 0.10 g in carbon dioxide-free water R, heating gently if
necessary, and dilute to 50 m L with the same solvent.
A p p e a ra n c e o f so lu tio n
Solution S is n o t more opalescent than reference
C^HaoClNO 347.9 1235-82-1 suspension II (2.2.1) and is colourless (2.2.2, Method II).
Temperature:
STORAGE
In an airtight container, protected from light.
IM P U R IT IE S
Specified impurities A , B, C, F.
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). I t is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances fa pharmaceutical use): D, E.
1-298 Bisacodyl 2016
Reference solutions Prepare the reference solutions using silver C. It gives the reaction o f nitrates (2.3.1).
standard solution (5 ppm Ag) R and diluting with a D. p H (2.2.1): maximum 2.0 for solution S2 (see Tests).
6.5 per cent V/V solution of lead-free nitric acid R. TESTS
Source Silver hollow-cathode lamp. S o lu tio n S I
Wavelength 328.1 nm. Shake 5.0 g by gently heating in 10 m L of water R and add
Atomisation device Air-acetylene flame. 20 m L o f nitric add R. H eat until dissolution, cool and dilute
to 100 m L with water R.
S u b sta n c e s n o t p re c ip ita te d b y am m onia
M aximum 1.0 p er c e n t S o lu tio n S2
Place 1.00 g in a 20 m L volumetric flask and add 2.0 m L of
In a porcelain o r quartz dish, ignite 2.0 g, increasing the
lead-free nitric add R. Allow a d d attack to take place w ithout
tem perature very gradually to 600 ± 50 °Cj allow to cool.
h eatin g and if necessary warm slightly at the end to
M oisten the residue with 2 m L o f nitric acid R, evaporate to
completely dissolve the test sample. Add 10 m L of water R,
dryness on a w ater-bath and carefully heat and ignite once
shake and add, in small fractions, 4.5 m L of lead-free
more at 600 ± 50 °C. After cooling, dissolve the residue in
ammonia R; shake and allow to cool. Dilute to 20.0 m L with
5 m L of nitric acid R and dilute to 20 m L with water R
water R, shake again and allow the solids to settle. The clear
T o 10 m L of this solution, add concentrated ammonia R until
supernatant solution is solution S2.
alkaline and filter. W ash the residue with water R and
evaporate the com bined filtrate and washings to dryness on a A cid ity
water-bath. A dd 0.3 m L of dilute sulfuric acid R and ignite. Suspend 1.0 g in 15 m L of water R and shake several times.
T h e residue weighs a maximum o f 10 mg. Allow to stand for 5 min and filter. T o 10 m L o f the filtrate,
add 0.5 m L o f phenolphthalein solution R l. N o t more than
L oss o n d ry in g (2.2.32)
0.5 m l. o f 0.1 M sodium hydroxide is required to change the
M axim um 7.0 p er cent, determined on 1.000 g by drying in
colour of the indicator to pink.
an oven at 105 °C for 3 h.
C h lo rid e s (2.4.4)
A SSA Y M aximum 200 ppm.
T o 0.300 g add 10 m L of a mixture o f equal volumes of
T o 5.0 m L of solution S 1, add 3 m L of nitric add R and
nitric add R and water R, heat to boiling and maintain boiling
dilute to 15 m L with water R.
for 2 m in. Add 0.1 g o f potassium chlorate R, heat to boiling
and maintain boiling for 1 min. Add 10 m L o f water R and C opper
heat until the solution becomes colourless. T o the hot M aximum 50 ppm .
solution, add 200 m L o f water R and 50 m g of xylenol orange Atomic absorption spectrometry (2.2.23, Method I).
triturate R. T itrate with 0.1 M sodium edetate until a yellow Test solution Solution S2.
colour is obtained.
Reference solutions Prepare the reference solutions using copper
1 m L of 0.1 M sodium edetate is equivalent to 20.90 m g o f Bi. standard solution (10 ppm Cu) R and diluting with a
STO R A G E 37 p er cent V/V solution o f lead-free nitric acid R.
Protected from light. Source Copper hollow-cathode lamp.
___________________________________________________________________________________________ PhEur Wavelength 324.7 nm.
Atomisation device Air-acetylene flame.
L ead
M aximum 20 ppm.
1-302 Bismuth Subsalicylate 2016
Atomic absorption spectrometry (2.2.23, MethodII). filtrate for identification test B. Wash the residue with dilute
Test solution Solution S2. hydrochloric acid R and then with water R. Dissolve the
residue in 0.5-1 m L of diluxe sodium hydroxide solution R.
Reference solutions Prepare the reference solutions using lead
Add 15 m L of water R. Neutralise with dilute hydrochloric
standard solution (10 ppm Pb) R and diluting with a
acid R. T he solution gives reaction (a) of salicylates (2.3.1).
37 per cent V/V solution o f lead-free nitric acid R.
B. T he filtrate obtained in identification test A gives
Source Lead hollow-cathode lamp.
reaction (b) of bismuth (2.3.1).
Wavelength 283.3 nm (depending on the apparatus, the line
at 217.0 nm may be used). TESTS
S o lu tio n S
Atomisation device Air-acetylene flame.
In a porcelain or quartz dish, ignite 1.0 g, increasing the
Silver temperature very gradually. H eat in a muffle furnace at
M aximum 25 ppm. 600 ± 25 °C for 2 h. Cool and dissolve the residue with
Atomic absorption spectrometry (2.2.23, Method I). warming in 4 m L of a mixture o f equal volumes of lead-free
Test solution Solution S2. nitric add R and water R and dilute to 20 m L with water R.
Reference solutions Prepare the reference solutions using silver A cidity
standard solution (5 ppm Ag) R and diluting with a Shake 2.0 g with 30 m L of ether R for 1 min and filter.
37 per cent V/V solution of lead-free nitric acid R. T o the filtrate add 30 m L o f alcohol R and 0.1 m L of thymol
Source Silver hollow-cathode lamp. blue solution R. N ot more than 0.35 m L of 0.1 M sodium
hydroxide is required to change the colour o f the indicator to
Wavelength 328.1 nm.
blue.
Atomisation device Air-acetylene flame.
C h lo rid e s (2.4.4)
S u b sta n c e s n o t p re c ip ita te d b y a m m o n i a Maximum 200 ppm.
M aximum 1.0 per cent.
Dissolve 0.250 g in a mixture of 2 m L o f nitric acid R, 5 m L
To 20 m L o f solution S i, add concentrated ammonia R until of water R and 8 m L o f methanol R.
an alkaline reaction is produced and filter. Wash the residue
N itra te s
with water R, and evaporate the combined filtrate and
Maximum 0.4 per cent.
washings to dryness on a water-bath. T o the residue, add
0.3 m L of dilute sulfuric acid R and ignite. T he residue weighs T o 0.1 g add 10 m L o f water R and, with caution, 20 m L of
a maximum of 10 mg. sulfuric acid R and stir. The solution is not more intensely
yellow coloured than a reference solution prepared at the
Loss o n d ry in g (2.2.32)
same time using 0.1 g o f salicylic add R, 6 m L o f water R,
M aximum 3.0 per cent, determined on 1.000 g by drying in
4 m L of nitrate standard solution (100 ppm NO 3 ) R and
an oven at 105 °C.
20 m L o f sulfuric acid R.
A SSAY C opper
Dissolve with heating 0.250 g in 10 m L of a mixture of Maximum 50 ppm.
2 volumes o f perchloric add R and 5 volumes of water R.
Atomic absorption spectrometry (2.2.23, Method I).
T o the hot solution, add 200 m L o f water R and 50 m g o f
xylenol orange triturate R. Titrate with 0.1 M sodium edetate Test solution Solution S.
until a yellow colour is obtained. Reference solutions Prepare the reference solutions using copper
1 m L o f 0.1 M sodium edetate is equivalent to 20.90 m g of Bi. standard solution (10 ppm Cu) R and diluting with a
6.5 per cent V/V solution of lead-free nitric acid R.
PhEur
Source Copper hollow-cathode lamp.
Wavelength 324.7 nm.
Atomisation device Air-acetylene flame.
Bismuth Subsalicylate ***** L ead
★ ★ M aximum 20 ppm.
(Ph Eur monograph 1495) *
Atomic absorption spectrometry (2.2.23, Method II).
C7H 5B i04 362.1 14882-18-9 Test solution Solution S.
PhEur__________________________________________________________
Reference solutions Prepare the reference solutions using lead
D E F IN IT IO N standard solution (10 ppm Pb) R and diluting with a
Complex of bismuth and salicylic add. 6.5 per cent V/V solution of lead-free nitric add R.
C o n te n t Source Lead hollow-cathode lamp.
56.0 per cent to 59.4 per cent of Bi (At 209.0) (dried Wavelength 283.3 nm (depending on the apparatus, the line
substance). at 217.0 nm may be used).
CHA RACTERS Atomisation device Air-acetylene flame.
A p p e a ra n c e S ilver
White or almost white powder. M aximum 25 ppm.
S olubility Atomic absorption spectrometry (2.2.23, Method I).
Practically insoluble in water and in alcohol. It dissolves in Test solution Solution S.
mineral acids with decomposition. Reference solutions Prepare the reference solutions using silver
ID E N T IF IC A T IO N standard solution (5 ppm Ag) R and diluting with a
A. T o 0.5 g add 10 m L of hydrochloric add R l. H eat on a 6.5 per cent V/V solution o f lead-free nitric add R.
boiling water-bath for 5 min. Cool and filter. Retain the Source Silver hollow-cathode lamp.
2016 Bisoprolol Fumarate 1-303
G. (2/?S)-l-[4-
A. (2i?5)-1-(4-hydroxymethyl-phenoxy)-3- [ [(2-isopropoxyethoxy)methoxy] methyl] phenoxy] -3-
isopropylaminopropan-2-ol, isopropylaminopropan-2-ol,
H OH H OH
n ^ch 3
ch3
ch3
/\^ C H 3 and enantiomer
O CH3 and enantiomer
H OH ***
Bleomycin Sulfate ★ ★
★ ★
Bleomycin Sulphate *****
OHC and enantiomer
(Ph Eur monograph 0976)
L. 4-[[(2i?S)-2-hydroxy-3-
(isopropylamino)propyl] oxy] benzaldehyde,
O^ ,NH
V H« n3>-—s. ji y—s
H,N
\ __ I > H -y ^ N H N_ /
/ ~ \ . nh k .x y
^ O CH3 and enantiomer
*n3
, x H2S 04
N . (2JRS)-1 - [4- [(2-ethoxyethoxy)methyl] phenoxy] -3-
isopropylaminopropan-2-olj
hv 0H «
ch 3
bleomycin B2:
R = NH-[CH2]4-NH-C(=NH)-NH2
'OCH3 and enantiomer
Q. (2i?S)-l-(isopropylamino)-3-[4- 9041-93-4
(2-methoxyethoxy)methyl]phenoxypropan-2-ol,
A ctio n a n d u se
Cytotoxic antibacterial.
H OH
H
n^ ch3 P re p a r a tio n
Bleomycin Injection
ch 3
h3c and enantiomer PhEur.
D E F IN IT IO N
R. (2i?S)-l-(isopropylamino)-3-(4-methylphenoxy)propan-2-
Sulfate of a mixture of glycopeptides produced by
ol,
Streptomyces verdciUiis or by any other means; the 2 principal
components of the mixture are N-[3-
OH
(dimethylsulfonio)propyl]bleomycinamide (bleomycin A 2)
and N- [4-(carbamimidoylamino)butyI] bleomycinamide
OHC (bleomycin B2).
P o te n c y
S. 4-hydroxybenzaldehyde, Minimum 1500 IU/m g (dried substance).
CHARACTERS
A p p e a ra n c e
0"\ 1h 3
,c
W hite or yellowish-white, very hygroscopic powder.
ch 3 S o lu b ility
OHC
Very soluble in water, slightly soluble in anhydrous ethanol,
practically insoluble in acetone.
T . 4- [(3-isopropyl-2-oxo-1,3-oxazolidin-5- ID E N T IF IC A T IO N
yl) methoxy] benzaldehyde. A. Examine the chromatograms obtained in the test for
composition.
Results T he 2 principal peaks in the chromatogram obtained
o-"\ <3
ch with the test solution are similar in retention time and size to
> ^ L y N~< the 2 principal peaks in the chromatogram obtained with
ch3 reference solution (a).
HO
B. It gives the reactions o f sulfates (2.3.1).
TESTS
U . 5- [ [4-(hydroxymethyl)phenoxy] methyl] -3-isopropyl-1,3-
A p p e a ra n c e o f so lu tio n
oxazolidin-2-one.
T h e solution is clear (2.2.1) and its absorbance (2.2.25) at
PhEur 430 nm is not greater than 0.10.
Dissolve 0.200 g in water R and dilute to 10.0 m L with the
same solvent.
p H (2.2.3)
4.5 to 6.0.
1-306 Bleomycin Sulfate 2016
Dissolve 50 m g in carbon dioxide-free water R and dilute to Atomisation (iedce Air-acetylene flame.
10 m l. with the same solvent. L oss o n d ry in g (2.2.32)
C o m p o sitio n M aximum 3.0 per cent, determ ined on 50 mg by drying at
Liquid chrom atography (2.2.29): use the normalisation 60 °C at a pressure not exceeding 0.67 kPa for 3 h.
procedure. B a c te ria l e n d o to x in s (2.6.14)
Test solution Dissolve 25 .0 mg o f the substance to be Less than 5 IU/m g, if intended for use in the manufacture of
examined in water R a n d dilute to 50.0 m L with the same parenteral preparations without a further appropriate
solvent. procedure for the removal o f bacterial endotoxins.
Reference solution (a) Dissolve the contents of a vial of A SSAY
bleomycin sulfate CRS in water R and dilute to 10.0 m L with Carry out the microbiological assay o f antibiotics (2.7.2),
the same solvent. using the diffusion m ethod. Use bleomycin sulfate CRS as the
Reference solution (b) D ilute 1.5 m L of reference solution (a) chemical reference substance.
to 100.0 m L with water R.
STO RA G E
Column:
In an airtight container, at a tem perature o f 2 °C to 8 °C.
— sizer. I = 0.25 m , 0 = 4.6 mm;
If the substance is sterile, store in a sterile, airtight, tam per
— stationary phase: octadecylsifyl silica gel for chromatograph R
p roof container.
(7 pm).
Mobile phase: IM P U R IT IE S
— mobile phase A: methanol R; Specified impurities D
— mobile phase B: dissolve 0.960 g o f sodium Other detectable impurities (the following substances would, if
pentanesulfonate R in 900 m L o f acetic a d d (4.8 g/L present at a suffident levd, be detected by one or other o f
C 2H 4O 2), add 1.86 g o f sodium edetate R, dilute to the tests in the monograph. They are limited by the general
1000 m L with the same solvent and adjust to p H 4.3 acceptance criterion for other/unspecified impurities and/or
with ammonia R] by the general monograph Substances for pharmaceutical use
(2034). It is therefore no t necessary to identify these
impurities for dem onstration o f compliance. See also 5.10.
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V) Control of impurities in substances for pharmaceutical use): A , B,
0 - 60 10 ->40 90 —» 60 C.
60 - end 40 60
★ ★ LA B E LL IN G
Refined Borage Oil ★ ★
T h e label states, where applicable, that the oil is suitable for
*****
Refined Starflower Oil use in the manufacture of parenteral preparations.
(Refined Borage (Starflower) OH, Ph Eur monograph 2105) ------------------------------------------------------------------------------------- -- PhEur
PhEur_________ _________________________________________________
D E F IN IT IO N
Fatty oil obtained firom seeds of Borago officinalis L. ***
by extraction and/or expression. It is then refined. A suitable Borax *
★
*
★
antioxidant may be added. Sodium Borate; Sodium Tetraborate *****
CHARACTERS (Ph. Eur. monograph 0013)
A p p e a ra n c e N a2B40 7,10H20 . 381.4 1303-96-4
CleaTj light yellow or yellow liquid.
PhEur_________________________________
S olubility
Practically insoluble in water and in ethanol (96 per cent), D E F IN IT IO N
misrible with light petroleum. Disodium tetraborate decahydrate.
R elativ e d en sity C o n te n t
About 0.921. 99.0 per cent to 103.0 per cent o f N a 2B407, 10H 20 .
Refractive index A bout 1.476. CHARACTERS
ID E N T IF IC A T IO N A p p e a ra n c e
First identification B W hite or almost white, crystalline powder, colourless crystals
or crystalline masses, efflorescent.
Second identification A
S olu b ility
A. Identification of fatty oils by thin-layer chromatography
Soluble in water, very soluble in boiling water, freely soluble
(2.5.2).
in glycerol.
Results T he chromatogram obtained is similar to the
corresponding chromatogram shown in Figure 2.3.2.-1. ID E N T IF IC A T IO N
A. T o 1 m L o f solution S (see Tests) add 0.1 m L of svifunc
B. Composition o f fatty adds (see Tests).
add R and 5 m L o f methanol R and ignite. The flame has a
TESTS green border.
A cid valu e (2.5. 1) B. T o 5 m L o f solution S add 0.1 m L of phenolphthalein
Maximum 0.5, or maximum 0.3 if intended for use in the solution R. T he solution is red. O n the addition of 5 m L o f
manufacture of parenteral preparations. glycerol (85 per cent) R the colour disappears.
P e ro x id e v alu e (2.5.5, Method A) C. Solution S gives the reactions of sodium (2.3.1).
M aximum 10.0, or maximum 5.0 if intended for use in the
TESTS
manufacture o f parenteral preparations.
S o lu tio n S
U n sap o n ifiab le m a tte r (2.5.7)
Dissolve 4.0 g in carbon dioxide-free water R prepared from
M aximum 2.0 per cent, determined on 5.0 g.
distilled water R and dilute to 100 m L with the same solvent.
A lkaline im p u ritie s (2.4.19)
A p p e a ra n c e o f so lu tio n
It complies with the test.
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
C o m p o sitio n o f fa tty a d d s ( 2.4.22, Method A)
p H (2.2.3)
Use the mixture of calibrating substances in Table 2.4.22.-3. 9.0 to 9.6 for solution S.
Composition of the fatty-acid fraction of the oil:
S u lfates (2.4.13)
— saturated fatty acids of chain length less than C;<j: maximum
M aximum 50 ppm , determined on solution S.
0.3 per cent,
— palmitic acid: 9.0 per cent to 12.0 per cent, Use in this test 1.0 m L o f acetic acid R. Prepare the standard
— palmitoleic acid: maximum 0.6 per cent, using a mixture o f 3 m L of sulfate standard solution
— stearic acid: 2.0 per cent to 6.0 per cent, (10 ppm SO4) R and 12 m L of distilled water R.
— oleic acid: 12.0 per cent to 22.0 per cent, A m m o n iu m (2.4.1)
— linoleic acid: 30.0 per cent to 41.0 per cent, M aximum 10 ppm.
— gamma-Unolenic acid: 17.0 per cent to 27.0 per cent, Dilute 6 m L o f solution S to 14 m L with water R. Prepare
— alpha-linolenic acid: maximum 0.5 per cent, the standard using a mixture o f 2.5 m L of ammonium
— arachidic acid: maximum 0.5 per cent, standard solution (1 ppm N H J R and 7.5 m L of water R.
— eicosenoic acid: 2.8 per cent to 4.4 per cent,
A rse n ic (2.4.2, Method A)
— erucic acid: maximum 3.0 per cent,
Maximum 5 ppm , determined on 5 m L o f solution S.
— nervonic acid: maximum 4.5 per cent.
C a lc iu m (2.4.3)
B ra ssic a ste ro l (2.4.23)
M aximum 100 ppm , determined on solution S.
Maximum 0.3 per cent in the sterol fraction of the oil.
Prepare the standard using a mixture of 6 m L o f calcium
W a te r (2.5.32)
standard solution (10 ppm Ca) R and 9 m L o f distilled water R.
M aximum 0.1 per cent, determined on 1.00 g.
H eav y m e ta ls (2.4.8)
STO RA G E M aximum 25 ppm.
U nder an inert gas, in a well-filled, airtight container,
12 m L of solution S complies w ith test A. Prepare the
protected from light.
reference solution using lead standard solution (1 ppm Pb) R.
1-308 Boric Acid 2016
the num ber o f animals used, or refine or replace the test proteases. T h e nicked protein is a fully active double-chain
procedure with the goal of prom oting animal welfare. polypeptide consisting o f a heavy chain (100 000 relative
T he potency of the reconstituted product is determined by molecular mass) and a light chain (50 000 relative molecular
an L D 50 assay in mice or by a m ethod validated with respect mass), connected by a disulfide bond.
to the L D 50 assay. T h e potency is expressed in terms of the P R O D U C T IO N
L D 50 for mice or relative to the reference preparation. GENERAL PROVISIONS
For determ ination of the LDsoj graded doses of the product Production of the toxin is based on seed cultures, managed
are injected intraperitoneally into groups o f mice and the in a defined seed-lot system in which the ability to produce
L D 50 is calculated by the usual statistical methods (5.3) from toxin is conserved. T h e production m ethod m ust be shown
the mouse lethality in each group. A suitable reference to yield consistently product o f activity and profile
preparation is assayed in parallel; the potency of the toxin is comparable to that of lots shown in clinical studies to be of
expressed relative to the reference or the value found for the adequate safety and efficacy.
reference is within suitable limits defined in terms of the T h e production m ethod is validated to dem onstrate that the
assigned potency. product, if tested, would comply with the general test of
After validation with respect to the L D 50 assay (reference abnorm al toxicity (2.6.9) using not less than the m axim um
m ethod), the product may also be assayed by other methods hum an clinical dose, in the presence o f a suitable am ount of
that are preferable in terms o f animal welfare, for example specific botulinum type B antitoxin used for neutralisation.
mouse bioassays using paralysis as the end-point, ex vivo T he production m ethod and stability o f th e finished product
assays using mouse phrenic nerve diaphragm, endopeptidase and relevant intermediates are evaluated using the tests
assays in vitro and cell-based assays. below. Such tests include the specific toxin activity per
For alternative replacement methods the potency is milligram o f protein o f purified toxin in an appropriate
calculated with respect to a suitable reference preparation functional model o f toxin activity and may be supported by
calibrated in mouse L D 50 units. tests confirming the presence o f botulinum toxin type B, and,
T he estim ated potency is n o t less than 80 per cent and not if appropriate, associated non-toxic proteins.
more than 125 per cent o f the stated potency. BACTERIAL SEED LOTS
T he confidence limits (P = 0.95) are n o t less than A highly toxigenic strain o f C. botulinum o f known toxin
80 per cent and not more than 125 p er cent of the estimated type B and confirmed absence o f genes encoding other
potency. botulinum toxins (particularly botulinum toxin types A
T he test may be repeated b u t when m ore than 1 test is and F ), with known origin and history, is grown using
performed, the results o f all valid tests m ust be combined in suitable media. T he bacterial strain, used for the m aster seed
the estimate o f potency. lot, shall be identified by historical records that include
information on its origin and the tests used to characterise
L A B E L L IN G
the strain. These will include morphological, cultural,
T he label states:
biochemical, genetic and serological properties o f the strain.
— the num ber of units of toxin per vial with a statem ent
T h e m aster seed lot and the working seed lot, where
that units are product specific and n o t applicable to other
applicable, m ust be dem onstrated to have identical profiles.
preparations containing botulinum toxin type A;
Only a seed lot that complies with the following requirem ents
— the nam e and the volume of the diluent to be added for
may be used.
reconstitution of the dried product.
Id e n tific a tio n
___________________________________________________________ PhEur
Each seed lot is identified as containing pure cultures of
C. botulinum type B bacteria with no extraneous bacterial or
fungal contamination.
M ic ro b ia l p u rity
Botulinum Toxin Type B for ***** Each seed lot complies with the requirem ents for absence o f
contaminating micro-organisms. T he purity of bacterial
Injection ***** cultures is verified by m ethods o f suitable sensitivity. These
(Ph Eur monograph 2581) may include inoculation into suitable media and examination
PhEur___________________________________________________________ of colony morphology.
D E F IN IT IO N P h e n o ty p ic p a ra m e te rs
Botulinum toxin type B for injection is a liquid preparation Each seed lot m ust have a known fatty a d d profile, sugar
containing purified botulinum neurotoxin type B, which may fermentation profile (glucose, lactose, m annose, etc.) and
be present in the form of a complex w ith haemagglutinins proteolytic activity and m ust demonstrate relevant lipase,
and non-toxic proteins. Botulinum neurotoxin type B or its ledthinase and gelatinase activity.
haemagglutinin complex is prepared by a suitable purification G e n etic p u rity
process o f the liquid supernatant from a broth-culture o f a Each seed lot must have information on the toxin gene
suitable strain o f Clostridium botulinum type B. Suitable genomic location and on the toxin gene sequence, and
stabilisers may be added. comply with requirements for the absence of other genes
T he toxin is present in its native form as a complex o f encoding other toxin serotypes.
neurotoxin and non-toxin proteins and haemagglutinins with P ro d u c tio n o f activ e to x in
a total relative molecular mass o f approximately 700 000. A bacterial strain producing a high yield o f active toxin, as
T he neurotoxin is synthesised by the bacterium as a single determined by an acute toxidty assay, is suitable. Seed lots
chain polypeptide o f approximately 150 000 relative demonstrate a capability o f producing at least a m inim um
molecular mass that is activated during the fermentation toxidty level appropriate for the manufacturing process and
process via a proteolytic cleavage (nicking) by endogenous scale.
2016 Botulinum Toxin Type B for Injection 1-311
MANUFACTURER S REFERENCE PREPARATIONS Only a final lot that is within the limits approved for the
D uring development, reference preparations are established particular product and is satisfactory with respect to each of
for subsequent verification of batch consistency during the requirements given bdow un d er Identification, Tests and
production and for control of the bulk purified toxin and Assay may be released for use.
finished p ro d u ct T h ey are derived from representative p H (2.2.3)
batches of botulinum toxin type 8 that are characterised as T he p H of the product is within ± 0.5 p H units of the limit
described under Bulk Purified Toxin. approved for the particular product.
T he reference preparations are suitably characterised for their
ID E N T IF IC A T IO N
intended purpose and are stored in suitably sized aliquots
T he presence o f botulinum toxin type B is confirmed by a
under conditions ensuring their suitability.
suitable immunochemical m ethod (2.7.1).
BULK PURIFIED TOXIN
C. botulinum type B strain is grown anaerobically, in suitable TESTS
media, from which cultures are selected for step-up S te rility (2.6.1)
incubations under a suitably controlled anaerobic atmosphere It complies with the test for sterility.
through the seed culture and bulk fermentation stages to B a c te ria l e n d o to x in s (2.6.14)
allow maximum production of toxin. T he toxin is purified by Less than 10 IU per vial.
suitable methods to remove nucleic acids and components
ASSAY
likely to cause adverse reactions.
In accordance with the provisions o f the European
Only a purified toxin that complies with the following Convention for the Protection of Vertebrate Animals U sed
requirements may be used in the preparation o f the final for Experimental and O ther Scientific Purposes, tests must
bulk. F or each test and for each product, limits of acceptance be carried out in such a way as to use the minimum num ber
are established and each new purified toxin m ust comply of animals and to cause the least pain, suffering, distress or
with these limits. lasting harm. T h e L D 50 assay is associated with severe
R e sid u a l re a g e n ts suffering o f animals and manufacturers are strongly
Removal of residual reagents used in purification steps is encouraged to develop and validate assays that will reduce
confirmed by suitable limit tests or by validation of the the num ber of animals used, or refine or replace the test
process. procedure with the goal of prom oting animal welfare.
N ucleic a d d s T h e potency of the product is determ ined by an L D 50 assay
Removal of nucleic ad d s is confirmed by suitable limit tests in mice or by a method validated with respect to the L D 50
or by validation o f the process. assay. T he potency is expressed in terms o f the L D 50 for
Im m u n o lo g ic a l id e n tity mice or relative to the reference preparation.
T he presence of specific type B toxin is confirmed by a For determination of the L D 50, graded doses of the product
suitable immunochemical method (2.7.1). are injected intraperitoneally into groups o f mice and the
L D 50 is calculated by the usual statistical methods (5.5) from
Specific a ctiv ity
the mouse lethality in each group. A suitable reference
T he specific activity is confirmed in a mouse m o d d o f
preparation is assayed in parallel; the potency of the toxin is
toxidty or by in vivo/ex vivo methods validated with respect
expressed relative to the reference or the value found for the
to the L D 50 assay and expressed in mouse L D 50 units per
reference is within suitable limits defined in terms of the
milligram o f protein. Specific activity m ust not be less than
assigned potency.
1 x 108 mouse L D 50 units per milligram of protein.
After validation with respect to the L D 50 assay (reference
P ro te in
method), the product may also be assayed by other methods
T he total protein concentration is determined by a suitable
that are preferable in terms of animal welfare, for example
method. An acceptable value is established for the product
mouse bioassays using paralysis as the end-point, ex vivo
arid each batch m ust be shown to comply with the limits.
assays using mouse phrenic nerve diaphragm, endopeptidase
P ro te in pro file assays tn vitro and cell-based assays.
Identity and protein composition are determined by For alternative replacement methods the potency is
polyacrylamide g d electrophoresis (2.2.31) under reducing or calculated with respect to a suitable reference preparation
non-reducing conditions or by other suitable physicochemical calibrated in mouse L D 50 units.
m ethods such as size-exdusion chromatography (2.2.30),
T he estimated potency is not less than 80 per cent and not
comparing with suitable reference standards.
more than 125 p er cent of the stated potency.
T o ta l viab le c o u n t T he confidence limits (P = 0.95) are not less than
It complies with the limits approved for the particular 80 per cent and not more than 125 per cent of the estimated
product. potency.
FINAL BULK T he test may be repeated b u t w hen more than 1 test is
T he final bulk is prepared by adding approved exdpients to performed, the results o f all valid tests m ust be combined in
the bulk purified toxin. T h e solution is filtered through a the estimate of potency.
bacteria-retentive filter. If hum an albumin is added, it
L A B E LL IN G
complies with the monograph Human albumin
solution (0255). T he label states the num ber of units of toxin per vial with a
statement that units are product specific and not applicable
FINAL LOT to other preparations containing botulinum toxin type B.
T he final bulk is distributed aseptically into sterile, tam per
__________________________________________________________ PhEur
proof containers. Uniformity of fill is verified during filling
and the test for uniformity of content (2.9.6) is not required.
T he containers are d o sed so as to prevent contamination.
1-312 Bovine Serum 2016
T he animal health status of countries is defined by the where applicable, th at the serum has been inactivated and
‘Office International des Epizooties’ (OIE). the inactivation method;
F or serum to be subjected to a virus inactivation/removal where the serum has been inactivated by gamma
procedure, if evidence of viral contamination is found in any irradiation, the target minimum dose o f the irradiation
of the tests described above, the serum is acceptable only if procedure.
the virus is identified and shown to be present in an am ount PhEur
that has been shown in a validation study to be effectively
inactivated.
For serum that is not to be subjected to a virus
inactivation/removal procedure, if evidence of viral
contamination is found in any of the tests described above, Bretylium Tosiiate
the serum is not acceptable.
A test for bovine viral diarrhoea virus antibodies is carried •Br
out; an acceptance criterion for the titre is established taking s o 3-
account of the risk assessment.
Composition T h e content of a suitable selection o f the Me
following components is determined and shown to be within
the expected range for the type of serum: cholesterol, a-, p-
and y-globulin, albumin, creatinine, bilirubin, glucose, serum C18H 24BrN03S 414.4 61-75-6
aspartate transaminase (SAST, formerly SG O T - serum
glutamic-oxaloacetic transaminase), serum alanine A ctio n a n d u se
transaminase (SALT, formerly S G P T - glutamic-pyruvic Antiarrhythmic.
transaminase), phosphorus, potassium, calcium, sodium and P re p a ra tio n
pH . Bretylium Injection
T ests c a rr ie d o u t o n th e b a tc h p o s t-tr e a tm e n t
D E F IN IT IO N
If bovine viral diarrhoea virus was detected before virus
Bretylium Tosiiate is 2-bromobenzyl-N'-
inactivation/removal, the following test for bovine viral
ethyldimethylammonium-p-toluenesulfonate. It contains not
diarrhoea virus is carried out after virus inactivation/removal.
less than 99.0% and n o t more than 101.0% of
Test for bovine viral diarrhoea virus A validated test for bovine C i 8H 24BrN 0 3S, calculated with reference to the dried
viral diarrhoea virus is carried out, for example by inoculation substance.
into susceptible cell cultures, followed by n o t fewer than
3 subcultures and detection by immunostaining. N o evidence C H A R A C T E R IS T IC S
of the presence o f bovine viral diarrhoea virus is found. A white, crystalline powder. It melts at about 98°. It exhibits
polymorphism.
ID E N T IF IC A T IO N
Freely soluble in water, in ethanol (96%) and in methanol.
A. T he electrophoretic pattern corresponds to that for serum
and is consistent with the type (foetal or other) o f bovine ID E N T IF IC A T IO N
serum. A. T he infrared absorption spectrum, Appendix II A, is
B. Bovine origin is confirmed by a suitable immunochemical concordant with the reference spectrum o f bretylium tosiiate
m ethod (2.7.1). (RS 030). If the spectra are not concordant, dissolve a
quantity of the substance being examined in the minimum
TESTS
volume o f acetone by heating on a water bath at 50°,
O sm o lality (2.2.35) evaporate to dryness at room tem perature under a current of
280 mosmol/kg to 365 mosmol/kg for foetal bovine serum nitrogen and prepare a new spectrum of the residue.
and 240 mosmol/kg to 340 mosmol/kg for other types.
B. Carry out the method for thin-layer chromatography,
T o ta l p ro te in (2.5.33) Appendix HI A, vising the following solutions in water.
30 m g/m L to 45 mg/mL for foetal bovine serum and
(1) 0.5% wtv o f the substance being examined.
minim um 35 mg/mL for other types.
(2) 0.5% w/v of bretylium tosiiate BPCRS.
H a em o g lo b in
Maximum 4 mg/mL, determined by a validated method, CHROMATOGRAPHIC CONDITIONS
such as spectrophotometry. (a) Use a silica gel F2m precoated plate (M erck plates are
B a c te ria l e n d o to x in s (2.6.14) suitable).
Less than 10 IU /m L for donor bovine serum, less than (b) Use the mobile phase as described below.
25 IU /m L for foetal bovine serum, less than 100 IU /m L for (c) Apply 10 (iL of each solution.
other types. (d) Develop the plate to 15 cm.
S te rility (2. 6.1) (e) After removal of the plate, dry it in a current o f air and
It complies with the test. Use 10 m L for each medium. examine under ultraviolet light (254 nm).
M y co p lasm a s (2.6.7) MOBILE PHASE
It complies with the test. 15 volumes o f glacial acetic acid, 30 volumes of water and
STO RA G E 75 volumes o f butan-l-oL
Frozen at —10 °C or below. CONFIRMATION
L A B E L L IN G T he two principal spots in the chromatogram obtained with
T he label states: solution (1) correspond to those in the chromatogram
— the type o f serum; obtained with solution (2).
1-314 Brimonidine Tartrate 2016
TESTS IM P U R IT IE S
A cid ity
pH o f a 5.0% w hr solutionj 5.0 to 6.5} Appendix V L.
Clarity and colour o f solution
A 5.0% w/v solution is dear, Appendix IV A, and colourless, C O " *
Appendix IV B, M ethod II.
Related substances A. 2-bromobenzyldimethylamine,
Carry out the m ethod for liquid chromatography,
Appendix III D , using the following solutions in mobile
j* Ç j^ ^ N M e 2
phase.
(1) 0.20% w/v of the substance being examined.
(2) 0.002% w/v of the substance being examined. Br
(3) 0.05% w/v of bretytium tosilate BPCRS and 0.05% w/v o f
2-bromobenzyldimethylamine hydrochloride BPCRS. B. 3-bromobenzyldimethylamine,
CHROMATOGRAPHIC CONDITIONS
(a) U se a stainless steel colum n (25 cm x 4.6 mm) packed
with particles o f silica the surface o f which has been modified
by chemically bonded phenyl groups (5 (im) (Spherisorb » X T “ -
Phenyl is suitable).
(b) U se isocratic elution and the mobile phase described C. 4-bromobenzyldimethylamine.
below.
(c) U se a flow rate o f 2 m L p er minute.
(d) U se an am bient column tem perature.
(e) U se a detection wavelength of 265 nm. ★* * +★
Brimonidine Tartrate ★ ★
(f) Inject 20 (iL o f each solution. ★ ★
MOBILE PHASE
(Ph. Eur. monograph 2760)
0.5 volume o f triethylamine, 2 volumes o f glacial acetic add,
19 volumes o f acetonitrile and 81 volumes of 0.01m sodium,
o H OH
octanesulfonate. C 02H
SYSTEM SUITABILITY Y jc Y > • h<v >;Hx oOH
T he test is n o t valid unless, in the chromatogram obtained H
with solution (3), the resolution factor between the two
principal peaks is at least 6.0. C 15H 16BrN50 6 442.2 70359-46-5
LIMITS
A c tio n a n d u se
In the chrom atogram obtained with solution (1):
Alpha2-adrenoceptor agonist; treatm ent o f hypertension
the area of any secondary peak is not greater than half the area
of the peak in the chrom atogram obtained with solution (2) PhEur.
(0.5% ); DEFINITION
the sum of the areas o f all such peaks is not greater than the 5-Bromo-N-(imidazolidin-2-ylidene)qiiinoxalin-6-amine
area o f the principal peak in the chromatogram obtained with (2i?,3.R)-2j3-dihydroxybutanedioate.
solution (2) (1%).
C o n te n t
Disregard the peak due to tosilate (retention time, about 99.0 per cent to 101.0 per cent (dried substance).
2 minutes) and any peak with an area less than 0.05 times
the area of the principal peak in the chromatogram obtained CHARACTERS
with solution (2) (0.05%). A p p e a ra n c e
W hite or slightly yellowish or slightly brownish powder.
Loss on drying
W hen dried to constant weight over phosphorus pentoxide at S o lu b ility
60° a t a pressure not exceeding 0.7 kPa, loses not m ore than Soluble in water, practically insoluble in anhydrous ethanol
3.0% of its weight. U se 1 g. and in toluene.
Sulfated ash ID E N T IF IC A T IO N
N ot m ore than 0.1%, Appendix IX A. Use 1 g. A. Specific optical rotation (see Tests).
A SSA Y B. Infrared absorption spectrophotometry (2.2.24).
Dissolve 0 .2 g in 5 0 m L of 1,4-dioxan and carry out Comparison brimonidine tartrate CRS.
M ethod I for rton-aqueous titration, Appendix VHI A, using TESTS
0 .0 2 5 m perchloric add F S as titrant and determining the
S p ecific o p tic a l ro ta tio n (2.2.7)
end-point potentiometrically. Each m L of 0.025m perchloric + 9.0 to + 10.5 (dried substance).
add VS is equivalent to 1 0 .3 6 mg o f C ] 8H 24BrN 03 S.
Dissolve 0.50 g in water R and dilute to 50.0 m L with the
STO RA G E same solvent.
Bretylium Tosilate should be kept in an airtight container R e la te d su b s ta n c e s
and protected from lig h t Liquid chromatography (2.2.29).
2016 Brimonidine Tartrate 1-315
★* ★ Injection 20 nL.
★ ★
Bromazepam ★ ★
Run tone 4 times the retention tim e o f brom azepam .
(Ph Eur monograph 0879) *****
Identification of impurities U se the chrom atogram supplied
with bromazepam for system suitability CRS and the
chromatogram obtained w ith reference solution (b) to
identify the peaks due to impurities A, B, C , D and E.
Relative retention W ith reference to brom azepam (retention
time = about 5 min): im purity D = about 1.4;
impurity A = about 1.5; im purity C = about 1.6;
impurity E = about 2.1; im purity B = about 2.2.
System suitability: reference solution (b):
— resolution: minimum 4.0 betw een the peaks due to
Ci4H10BrN3O 316.2 1812-30-2 brom azepam and im purity D and m inim um 1.2 between
the peaks due to impurities A and C.
Action and use
Benzodiazepine.
Limits:
— correction factors: for the calculation o f content, multiply
PhEir. the peak areas of the following impurities by the
corresponding correction fa c to r impurity A = 1.3;
D E F IN IT IO N
impurity B = 1.8; im purity E = 2.1;
7-Brom o-5-(pyridin-2-yl)-1,3-dihydro-2/i-l ,4-benzodiazepin-
— impurities A , B, E: for each impurity, no t more than the
2-one.
area of the principal peak in the chrom atogram obtained
C o n te n t with reference solution (a) (0.1 per cent);
99.0 per cent to 101.0 per cent (dried substance). — unspecified impurities', for each impurity, n o t m ore than the
CHARACTERS area of the principal peak in the chrom atogram obtained
A p p e a ra n c e with reference solution (a) (0.10 per cent);
W hite or yellowish, crystalline powder. — total: n o t more than twice the area of the principal peak in
the chromatogram obtained with reference solution (a)
S o lu b ility
(0.2 per cent);
Practically insoluble in water, slighdy soluble or sparingly
— disregard limit. 0.5 times the area o f the principal peak in
soluble in ethanol (96 per cent) and in methylene chloride.
the chromatogram obtained with reference solution (a)
ID E N T IF IC A T IO N (0.05 per cent).
Infrared absorption spectrophotom etry (2.2.24). L o ss o n d ry in g (2.2.32)
Comparison bromazepam CRS. M axim um 0.2 per cent, determ ined on 1.000 g by drying at
80 °C at a pressure not exceeding 2.7 kPa for 4 h.
TESTS
R e la te d su b s ta n c e s S u lfa te d a s h (2.4.14)
L iquid chrom atography (2.2.29). Prepare the solutions M axim u m 0.1 per cent, determ ined on 1.0 g.
immediately before use. A SSA Y
Test solution Dissolve 10.0 m g o f the substance to be Dissolve 0.250 g in 20 m l . o f anhydrous acetic acid R
examined in 9 m L o f a mixture of 1 volume of acetonitrüe R Add 50 m l . o f acetic anhydride R. T itrate w ith 0.1 M
and 8 volumes o f methanol R Dilute to 20.0 m L with an perchloric acid, determining the end-point potentiometrically
11.33 g/L solution o f potassium àihydrogen phosphate R (2. 2. 20).
previously adjusted to p H 7.0 with a 100 g/L solution of 1 m L of 0.1 M perchloric add is equivalent to 31.62 m g
potassium hydroxide R. o f C i 4H 10B rN 3O.
Reference solution (a) D ilute 1.0 m L o f the test solution to
STORAGE
100.0 m L w ith the mobile phase. Dilute 1.0 m L o f this
solution to 10.0 m L with the mobile phase. Protected from light.
ID E N T IF IC A T IO N
First identification A , E.
Second identification B, C, D, E.
A. Infrared absorption spectrophotometry (2.2.24).
Comparison bromhexine hydrochloride CRS.
If the spectra obtained in the solid state show differences,
A. R = H : (2-amino-5-bromophenyl) (pyridin-2- dissolve the substance to be examined and the reference
yl)methanone} substance separately in methanol R, evaporate to dryness and
record new spectra using the residues.
B. R = C O -C H 2-C l: N - [4-bromo-2-(pyridin-2-
ylcarbonyl) phenyl] -2-chloroacetamide, B. Thin-layer chromatography (2.2.27).
E. R = C O -C H 2-Br: 2-bromo-N- [4-bromo-2-(pyridm-2- Test solution Dissolve 20 mg of the substance to be examined
ylcarbonyl)phenyl]acetamide, in methanol R and dilute to 10 m L with the same solvent.
Reference solution Dissolve 20 mg of bromhexine
hydrochloride CRS in methanol R and dilute to 10 m L with the
sam e solvent.
Plate TLC silica gel F254 plate R.
Mobile phase glacial acetic acid R, water R, butanol R
(17:17:66 VIVIV).
Application 20 |iL.
Development Over 3/4 of the plate.
C. 7-brom o-5-(6-m ethylpyridin-2-yl)-l,3-dihydro-2H -l,4-
benzodiazepin-2-one, Drying In air.
Detection Examine in ultraviolet light at 254 nm.
Results T h e principal spot in the chromatogram obtained w ith
the test solution is similar in position and size to the principal
spot in the chromatogram obtained with the reference
solution.
C. Dissolve about 25 m g in a mixture of 1 m L o f dilute
sulfuric acid R and 50 m L of water R. Add 2 m L o f methylene
chloride R and 5 m L of chloramine solution R and shake.
D . 3-am ino-6-bromo-4-(pyridin-2-yl)quinolin-2( \H)-one.
A brownish-yellow colour develops in the lower layer.
PhEur
D. Dissolve about 1 m g in 3 m L of 0.1 M hydrochloric acid.
T h e solution gives the reaction of primary aromatic amines
(2.3.1).
E. Dissolve about 20 mg in 1 m L o f methanol R and add
★ ★
Bromhexine Hydrochloride ★ ★ 1 m L of water R. T he solution gives reaction (a) o f chlorides
(2.3.1).
(Ph Eitr monograph 0706) *★ *
TESTS
Br A p p e a ra n c e o f so lu tio n
T h e solution is clear (2.2.1) and not more intensely coloured
fS -H3 . HCI
than reference solution Y 6 (2.2.2, Method II).
Dissolve 0.6 g in methanol R and dilute to 20 m L with the
nh2 same solvent.
R e la te d su b sta n c e s
Ci4H2iBr2ClN2 412.6 611-75-6 Liquid chromatography (2.2.29).
Test solution Dissolve 50 mg of the substance to be examined
A ctio n a n d u se in methanol R and dilute to 10.0 m L with the same solvent.
Mucolytic.
Reference solution (a) Dissolve 5 m g o f bromhexine
P h E tr ____________________ impurity C CRS in methanol R, add 1.0 m L of the test
solution and dilute to 10.0 m L with the same solvent.
D E F IN IT IO N
Reference solution (b) Dilute 1.0 m L of the test solution to
AT-(2-Amino-3,5-dibromobenzyi)-2V-methylcyclohexanamine
100.0 m L with methanol R. Dilute 1.0 m L of this solution to
hydrochloride.
10.0 m L with methanol R.
Content Column:
98.5 per cent to 101.5 per cent (dried substance). — size: I = 0.12 m, 0 = 4.6 mm,
CHARACTERS — stationary phase: end-capped octadecylsüyl silica gel for
Appearance chromatography R (3 nm).
W hite o r almost white, crystalline powder. Mobile phase Mix 0.50 m L of phosphoric acid R in 950 m L of
Solubility water R, adjust to p H 7.0 with triethylamine R (about
Very slightly soluble in water, slightly soluble in alcohol and 1.5 mL) and dilute to 1000 m L with water R,
in methylene chloride. mix 20 volumes o f this solution with 80 volumes of
acetonitrüe R.
It shows polym orphism (5.9).
1-318 Bromocriptine Mesilate 2016
n o t m ore than 1 such peak has an area greater than the 4,6,6a,7,8,9-hexahydroindolo[4,3-j&]quinoline-9-carboxamide
area of the principal peak in the chromatogram obtained ((95)-2-bromo-a-ergocriptine),
w ith reference solution (b) (0.1 p er cent);
— total: not more than 1.5 times the area of the principal
peak in the chrom atogram obtained with reference
solution (a) (1.5 p er cent);
— disregard limir. 0.5 times the area o f the principal peak in 'CO jH
the chrom atogram obtained with reference solution (b)
(0.05 per cent), apart from the peak due to impurity A.
L oss o n d ry in g (2.2.32)
D . (6aR,9i?)-5-bromo-7-methyi-4,6,6a,7,8,9-
M axim um 3.0 per cent, determ ined on 0.500 g by drying
hexahydroindolo [4,3-^] quinoline-9-carboxylic acid,
in vacuo at 80 °C for 5 h.
ASSAY
Dissolve 0.500 g in 80 m L o f a mixture of 10 volumes of
anhydrous acetic add R and 70 volumes of acetic anhydride R
Titrate with 0.1 M perchloric add, determining the end-point
potentiometrically (2.2.20).
1 m L of 0.1 M perchloric add is equivalent to 75.1 mg
of C 33H 44B rN 508 S.
E. (6aR,9/?)-5-bromo-7-methyl-4,6,6a,7,8,9-
STO RA G E hexahydroindolo [4,3-/^] quinoline-9-carboxamide,
In an airtight container, protected from light, at a
tem perature not exceeding —15 °C.
IM P U R IT IE S
Spedfied impurities A, B, C , D , E, F , G
F. (6aR,9i?)-5-bromo-N-[(25,55,10aS,10b5)-10b-hydroxy-2-
(l-m ethylethyl)-5-(2-methylpropyl)-3,6-dioxooctahydro-8/f-
oxazolo [3,2-a] pyrrolo [2,1 -c] pyrazin-2-yl] -7-methyl-
4,6,6a,7,8,9-hexahydroindolo[4,3-j^]quinoline-9-carboxamide
A. (6ai?,9i?)-5-bromo-N- [(2R,5S)-2-( 1-methylethyl)-5- ((2 '^ ^ -b ro m o -a-erg o crip tin e),
(2-methylpropyl)-3,6-dioxo-2,3,5,6,9,10-hexahydro-8/f-
oxazolo [3,2-a] pyirolo [2,1 -c] pyrazin-2-yl]-7-methyl-
4,6,6a,7,8,9-hexahydroindolo[4,3-/g]quinoline-9-carboxamide
(2-bromodehydro-a-ergocriptine),
C. (6ai?,95)-5-brom o-N-[(2i?,55,10a5,10b5)-lOb-hydroxy-2-
(1 -methylethyl) -5-(2-methylpropyl)-3 ,6-dioxooctahydro-8//-
oxazolo [3,2-a] pyrrolo [2, l-c]pyrazin-2-yl]-7-methyl-
2016 Bromperidol 1-321
Br
L oss o n d ry in g (2.2.32)
M axim um 0.5 per cent, determ ined on 1.000 g by drying in
an oven at 105 °C.
S u lfa te d a s h (2.4.14)
Maximum 0.1 per cent, determ ined on 1.0 g in a platinum
crudble.
Br
ASSA Y
Dissolve 0.300 g in 50 m L o f a mixture o f 1 volume o f
anhydrous acetic acid R and 7 volumes o f methyl ethyl ketone R. OH
T itrate with 0.1 M perchloric acidy using 0.2 m L of
naphthoJbenzein solution R as indicator.
E. 4-[4-(4-brom ophenyl)-4-hydroxypiperidin-l-yl]-l-[4-[4-
1 m L o f 0.1 M perchloric acid is equivalent to 42.03 mg (4-bromophenyl)-4-hydroxypiperidin-l-yl]phenyl]butan-l-
o f C 2iH 23B rFN 02. one,
STO RA G E
Protected from light.
IM P U R IT IE S
Specified impurities A , By C, D, E, F.
F . 4- [4- (4 '-bromobiphenyl-4-yl)-4-hydroxypiperidin-1-yl] -1 -
(4-fluorophenyl)butan-1-one.
OH PhEur
A. 1-(4-fluorophenyl)-4-(4-hydroxy-4-phenylpiperidin-1-
yI)butan-l-one,
★ ★
Bromperidol Decanoate ★ ★
Br
(Ph Eitr monograph 1397) *****
OH Br
B. 4- [4-(4-bromophenyl)-4-hydroxypiperidin- 1-yl]-1-
(2-fluorophenyl)butan-1-one, H,C
A c tio n a n d u se
D opam ine receptor antagonist; neuroleptic.
PhEur________________________________________
C . 4- [4-(biphenyl-4-yl)-4-hydroxypipeiidin- 1-yl] -1-
(4-fluorophenyl)butan-l-one, D E F IN IT IO N
4-(4-Brom ophenyl)-l-[4-(4-fluorophenyl)-4-
oxobutyl]piperidin-4-yl decanoate.
h 3c
C o n te n t
98.5 per cent to 101.0 per cent (dried substance).
'XV ~ Q J C r * OH
CHARACTERS
A p p e a ra n c e
W hite or almost white powder.
D . 4- [4-(4-bromophenyl)-4-hydroxypiperidin-1-yl] -1 -
S o lu b ility
(3-ethyl-4-fluorophenyl)butan- 1-one,
Practically insoluble in water, very soluble in methylene
chloride, soluble in ethanol (96 p er cent),
m p A bout 60 °C.
ID E N T IF IC A T IO N
A. Infrared absorption spectrophotom etry (2.2.24).
Comparison bromperidol decanoate CRS.
B. T o 0.1 g in a porcelain crucible add 0.5 g o f anhydrous
sodium carbonate R. H eat over an open flame for 10 min.
Allow to cool. T ake up the residue with 5 m L of dilute nitric
2016 Bromperidol Decanoate 1-323
acid, R and filter. T o 1 m L o f the filtrate add 1 m L of — disregard limit: 0.1 times the area of the principal peak in
water R. T h e solution gives reaction (a) of bromides (2.3.1). the chromatogram obtained with reference solution (b)
(0.05 per cent).
TESTS
A p p e a ra n c e o f so lu tio n L oss o n d ry in g (2.2.32)
T he solution is clear (2.2.1) and not m ore intensely coloured M aximum 0.5 p er cent, determined on 1.000 g by drying
than reference solution B 5 (2.2.2, Method II). m vacuo at 30 °C.
Dissolve 2.0 g in methylene chloride R and dilute to 20 mL S u lfa ted a s h (2.4.14)
with the same solvent. M aximum 0.1 p er cent, determined on 1.0 g in a platinum
crucible.
R e la te d su b sta n c e s
Liquid chromatography (2.2.29). Prepare the solutions ASSAY
immediately before use and protea from light. Dissolve 0.450 g in 50 m L of a mixture o f 1 volume of
Test solution Dissolve 0.100 g o f the substance to be anhydrous acetic acid R and 7 volumes of methyl ethyl ketone R.
examined in methanol R and dilute to 10.0 m L with the sam e T itrate with 0.1 M perchloric add, using 0.2 m L of
solvent. naphtholbenzein solution R as indicator.
Reference solution (a) Dissolve 2.5 mg o f bromperidol 1 m L o f 0.1 Mperchloric acid is equivalent to 57.46 mg
decanoate CRS and 2.5 mg o f haloperidol decanoate CRS in o f C 3iH 41B rFN 0 3.
methanol R and dilute to 50.0 m L w ith the same solvent. STORA GE
Reference solution (b) Dilute 5.0 m L of the test solution to Protected from light, at a temperature below 25 °C.
100.0 m L with methanol R. Dilute 1.0 m L o f this solution to
IM P U R IT IE S
10.0 m L with methanol R.
Specified impurities A, B, C , D, E, F , G, H , I, J, K
Column:
— size. / = 0,1 m , 0 = 4.0 mm; Other detectable impurities (the following substances would, if
— stationary phase: base-deactivated octadecylsUyl silica gel for present at a sufficient level, be detected by one o r other o f
chromatography R (3 |im). the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
Mobile phase:
by the general monograph Substances for pharmaceutical use
— mobile phase A: 27 g/L solution o f tetrabutylammonium
(2034). It is therefore n o t necessary to identify these
hydrogen sulfate R;
impurities for demonstration of compliance. See also 5.10.
— mobile phase B: acetonitrüe R;
Control of impurities in substances for pharmaceutical use): L.
3 0 -3 5 40 60
3 5 -4 0 40 ->80 60-> 20
O
Flow rate 1.5 mL/min.
A. 1-[4-(4-fluorophenyl)-4-oxobutyl]-4-phenylpiperidin-4-yl
Detection Spectrophotom eter at 230 nm .
decanoate,
Injection 10 |iL.
Relative retention W ith reference to brom peridol decanoate
(retention time = about 24 min): impurity G = about 0.10;
impurity L = about 0.15; impurity H = about 0.8;
im purity A = about 0.89; impurity I = about 0.91;
impurity B = about 0.96s haloperidol
decanoate = about 0.98; impurity F = about 1.10;
im purity C = about 1.15; impurity K = about 1.2 ; o
im purity E = about 1.23; impurity D = about 1.25.
System suitability: reference solution (a): B. 4-(4-brom ophenyl)-l-[4-(2-fluorophenyl)-4-oxobutyl]-
— resolution: minimum 1.5 between the peaks due to piperidin-4-yl decanoate,
haloperidol decanoate and bromperidol decanoate.
Limits:
— impurities A , B, C, D, E, F, G, H, I, J, K, for each
im purity, n o t more than the area o f the principal peak in
the chrom atogram obtained with reference solution (b)
(0.5 p er cent);
— unspecified impurities: for each impurity, not m ore than
0.2 times the area o f the principal peak in the O
chrom atogram obtained with reference solution (b)
( 0.10 per cent); C . 4-(4-brom ophenyl)-l-[4-(3-ethyl-4-fluorophenyl)-4-
— total: not more than 3 times the area of the principal peak oxobutyl]-piperidin-4-yl decanoate,
in the chromatogram obtained with reference solution (b)
(1.5 per cent);
1-324 Brompheniramine Maleate 2016
Br
Br
I. 4-(4-bromophenyl)-1- [4-(4-fluorophenyl)-4-
oxobutyl]piperidin-4-yl nonanoate,
J. 4-(4-bromophenyl)-l-[4-(4-fluorophenyl)-4-
oxobutyl]piperidin-4-yl undecanoate,
E. 4-(4'-bromobiphenyl-4-yl)-l-[4-(4-fluorophenyl)-4-
oxobutyl]piperidin-4-yl decanoate,
ch3
L. l-(4-fluorophenyl)ethanone.
F. 4-(biphenyl-4-yl)-1- [4-(4-fluorophenyl)-4- PhEur
oxobutyl]piperidin-4-yl decanoate,
★ ★
Brompheniramine Maleate ★ ★
(Ph. Eur. monograph 0977)
c o 2h
G. 4- [4-(4-bromophenyl)-4-hydroxypiperidin-1-yl] -1 -
(4-fluorophenyl)butan-1-one (bromperidol), and enantiomer
■( C02H
DEFINITION
(3/?5)-3-(4-Bromophenyi)-J\yV-dimethyl-3-(pyridin-2-
yl)propan-l-am ine (Z)-butenedioate.
Content
98.0 per cent to 101.0 per cent (dried substance).
2016 1-325
of silver nitrate required. Each m L o f 0.1m silver nitrate VS is mobile phase B: mix 25 volumes of a 2 g/L solution of
equivalent to 20.00 mg of C 3H 6B rN 0 4. sodium heptanesulfonate R and 75 volumes of acetonitrile R',
STORAGE
Bronopol should be protected from light. Time Mobile phase A Mobile phase B
(min) (per cent V/V) (percent V/V)
0 -4 63 37
A. R1 = C H 3j R2 = H: 4-(2-chlorophenyl)-9-methyl-6H- T h e test is not valid unless the chrom atogram obtained with
thieno[3j2-/| [ 1,2,4] triazolo [4,3-a] [l,4]diazepine solution (4) closely resembles the chromatogram supplied
(desbromobrotizolam), with buclizine hydrochloride impurity standard BPCRS.
D E F IN IT IO N
Buclizine Hydrochloride is (i?5)-l-(4-terr-butylbenzyl)-4-
(4-chlorobenzhydryl)piperazine dihydrochloride. It contains
Budesonide *****
n o t less than 99.0% and not more than 100.5% o f
*+ +*
C 28H 33C 1N 2, 2H C 1, calculated with reference to the dried (Ph. Evr. monograph 1075) *
substance.
C H A R A C T E R IS T IC S o P H
A white or slightly yellowish, crystalline powder.
Practically insoluble in water, sparingly soluble in propane-1,2- Ch J h H and epimer at C*
diok very slightly soluble in ethanol (96%).
ID E N T IF IC A T IO N
A. T h e infrared absorption spectrum, Appendix II A, is
concordant with the reference spectrum o f buclizine
hydrochloride (RS 032). C 25H340 6 430.5 51333-22-3
B. A 0.25% w/v solution in ethanol (50%) yields reaction A
characteristic o f chlorides, Appendix VI. A ctio n a n d use
Glucocorticoid.
TESTS
P re p a r a tio n s
R e la te d su b sta n c e s
Budesonide Aqueous Nasal Spray
C arry out the m ethod for liquid chromatography,
Appendix IQ D , using four solutions in the initial mobile Budesonide Inhalation Powder
phase containing (1) 0.0010% w/v o f the substance being Budesonide Inhalation Powder, pre-dispensed
examined, (2) 0.50% w/v o f the substance being examined, Budesonide Nebuliser Suspension
(3) 0.0010% w/v of Budesonide Pressurised Inhalation
l,4-bis(4-chbrobenzhy<byl)piperazine BPCRS and (4)
0.50% w/v o f buclizine hydrochloride impurity standard BPCRS. PhEur__________________________________________________________
Content TESTS
97.5 per cent to 102.0 per cent (dried substance). Related substances
CHARACTERS Liquid chromatography (2.2.29). Carry out the test protected
from light.
Appearance
W hite or almost white, crystalline powder. Solvent mixture acetomtrüe R, phosphate buffer solution pH 3.2 R
(32:68 VIV).
Solubility
Practically insoluble in water, freely soluble in methylene Test solution (a) Dissolve 50 mg of the substance to be
chloride, sparingly soluble in ethanol (96 p er cent). examined in 15 m L of acetonitrüe R and dilute to 50 m L with
phosphate buffer solution pH 3.2 R.
IDENTIFICATION
Test solution (b) Dissolve 25.0 mg o f the substance to be
First identification A. examined in 15 m L of acetomtrüe R and dilute to 50.0 m L
Second identification B, C, D. with phosphate buffer solution pH 3.2 R.
A. Infrared absorption spectrophotometry (2.2.24). Reference solution (a) Dilute 1.0 m L o f test solution (a) to
Comparison budesonide CRS. 10.0 m L with the solvent mixture. D ilute 1.0 m L of this
B. Thin-layer chromatography (2.2.27). solution to 100.0 m L with the solvent mixture.
Solvent mixture methanol R, methylene chloride R (10:90 VIV). Reference solution (b) Dissolve 5 m g o f budesonide for system
Test solution Dissolve 25 m g o f the substance to be examined suitabüity CRS (containing impurities A, D , G, K and L) in
in the solvent mixture and dilute to 10 m L with die solvent 1.5 m L of acetomtrüe R and dilute to 5 m L with phosphate
mixture.
buffer solution pH 3.2 R.
Reference solution (a) Dissolve 25 mg o f budesonide CRS in the Reference solution (c) Dissolve 25.0 mg o f budesonide CRS in
solvent mixture and dilute to 10 m L with the solvent 15 m L o f acetonitrüe R and dilute to 50.0 m L with phosphate
mixture.
buffer solution pH 3.2 R.
Column:
Reference solution (b) Dissolve 12.5 mg o f triamcinolone
— size: I — 0.15 m , 0 = 4.6 mm;
acetordde CRS in reference solution (a) and dilute to 5 m L
— stationary phase: end-capped octadecylsüyl silica gel for
with reference solution (a).
chromatography R (3 |im);
Plate TLC silica gel F2 5 4 plate R. — temperature: 50 °C.
Mobile phase Add a mixture o f 1.2 volumes o f water R and Mobile phase:
8 volumes of methanol R to a mixture o f 15 volumes of — mobile phase A: anhydrous ethanol R, acetonitrüe R,
ether R and 77 volumes of methylene chloride R. phosphate buffer solution pH 3.2 R (2:32:68 VIVIV)\
Application 5 jiL. — mobile phase B: acetomtrüe R, phosphate buffer solution
Development Over a path o f 15 cm. pH 3.2 R (50:50 VIV)-,
Drying \n air.
Detection A Examine in ultraviolet light at 254 nm. Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
Results A T h e principal spot in the chromatogram obtained 100 0
0 - 38
with the test solution is similar in position and size to the
principal spot in die chromatogram obtained with reference 3 8 -5 0 100 -»0 0 * 100
solution (a). 5 0 -6 0 0 100
Detection B Spray with alcoholic solution of sulfuric acid R; heat
at 120 °C for 10 m in or until the spots appear and allow to
Flow rate 1 mL/min.
cool; examine the chromatograms in daylight and in
ultraviolet light at 365 nm. Detection Spectrophotom eter at 240 nm.
Results B T he principal spot in the chromatogram obtained Injection 20 jiL o f test solution (a) and reference solutions (a)
with the test solution is similar in position, colour in daylight, and (b).
fluorescence in ultraviolet light at 365 nm and size to the Identification of impurities Use the chrom atogram supplied
principal spot in the chromatogram obtained with reference with budesonide for system suitability CRS and the
solution (a). chromatogram obtained with reference solution (b) to
System suitability: reference solution (b): identify the peaks due to impurities A, D , G , K and L.
— the chromatogram shows 2 clearly separated spots. Relative retention W ith reference to budesonide epimer B
C. Dissolve about 2 m g in 2 m L o f sulfuric add R. W ithin (retention time = about 17 min): impurity A = about 0.1;
5 min a yellow colour develops. W ithin 30 min the colour epimers o f impurity D = about 0.63 and 0.67;
changes to brown or reddish-brown. Cautiously add the impurity L = about 0.95; epimers o f im purity G = about 1.2
solution to 10 m L of water R and mix. T h e colour fades and and 1.3; epimers of impurity K = about 2.9 and 3.0.
a clear solution remains. System suitability: reference solution (b):
D . Dissolve about 1 mg in 2 m L o f a solution containing 2 g — peak-to-vaEey ratio: minimum 2.5, where Hp = height
o f phosphomdybdic add R dissolved in a mixture o f 10 m L of above the baseline of the l n o f the 2 peaks due to
dilute sodium hydroxide solution R, 15 m L o f water R and impurity G and Hv = height above the baseline of the
25 m L o f glacial acetic acid R. H eat for 5 m in on a water- lowest point o f the curve separating this peak from the
bath. Cool in iced water for 10 min and add 3 m L o f dilute peak due to budesonide epimer A (the 2nd of the
sodium hydroxide solution R. T h e solution is blue. 2 principal peaks); and minimum 3, where Hp = height
above the baseline o f the peak due to impurity L and
Hv = height above the baseline o f the lowest point o f the
curve separating this peak from the peak due to
budesonide epimer B (the 1st of the 2 principal peaks).
1-330 Budesonide 2016
Limits: IM P U R IT IE S
— correction factors: for the calculation of content, multiply Specified impurities A , D, K, L.
the peak areas o f the following impurities by the Other detectable impurities (the following substances would, if
corresponding correction facto r impurity D = 1.8; present at a sufficient level, be detected by one or other of
impurity K = 1.3; the tests in the monograph. They are limited by the general
— impurities A , L: for each impurity, n o t more than twice acceptance criterion for other/unspecified impurities and/or
the sum of the areas of the 2 peaks due to the budesonide by the general monograph Substances for pharmaceutical use
epimers in the chrom atogram obtained with reference (2034). It is therefore not necessary to identify these
solution (a) (0.2 p er cent); impurities for demonstration of compliance. See also 5.10.
— impurities D, K. for each impurity, for the sum of the Control of impurities in substances for pharmaceutical use): B, C,
areas of the 2 epim er peaks, not m ore than twice die sum E, F, G, H, I, J.
o f the areas of the 2 peaks due to the budesonide epimers
in the chrom atogram obtained with reference solution (a)
(0.2 per cent);
— unspecified impurities: for each individual peak, not more
than the sum o f the areas o f the 2 peaks due to the
budesonide epimers in the chromatogram obtained with
reference solution (a) (0.10 per cent);
— total: not more than 5 times the sum o f the areas o f the
2 peaks due to the budesonide epimers in the
chromatogram obtained with reference solution (a) A. 1 ip,16a,17,21-tetrahydroxypregna-l,4-diene-3,20-dione,
(0.5 per cent);
— disregard limit: 0.5 times the sum of the areas o f the
2 peaks due to the budesonide epimers in the
chromatogram obtained with reference solution (a)
(0.05 per cent).
E p im e r A
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modifications.
Mobile phase: B. R = H: 16a,17-[(LRS)-ethylidenebis(oxy)]-lip,21-
dihydroxypregna-1,4-diene-3,20-dione,
22 - 31 0 100
Bufexamac ★ ★
★ ★
(Ph. Eur. monograph 1179) *****
„0 H and epimer at C*
H
DEFINITION
2-(4-Butoxyphenyl)-N-hydroxyacetamide.
Content
H . 16a,17-[(li?5)-butylidenebis(oxy)]-21- 98.5 per cent to 101.5 per cent (dried substance).
hydroxypregnal,4,9(l l)-triene-3j20-dionej CHARACTERS
Appearance
W hite or almost white, crystalline powder.
Solubility
Practically insoluble in water, soluble in dimethylformamide,
slightly soluble in ethyl acetate and in methanol.
IDENTIFICATION
First identification B
Second identification A , C
I. 11P, 17,2 1-trihydroxy-3,20-dioxopregna-1,4-dien-16a-yl
A. Ultraviolet and visible absorption spectrophotometry
butanoate,
(2.2.25).
Test solution Dissolve 20 mg in methanol R and dilute to
20 m L with the same solvent. D ilute 1 m L of the solution to
--0 ^ \ ^ c h 3 50 m L with methanol R.
..q H and epimer at C* Spectral range 210-360 nm.
H Absorption maxima At 228 nm, 277 n m and 284 nm.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison bufexamac CRS.
J. 16a,17-[(liiS)-butylidenebis(oxy)]-9a-brom o-l ip ,2 1 - C. Thin-layer chromatography (2.2.27).
dihydroxypregna-1,4-diene-3,20-dione, Test solution Dissolve 10 mg of the substance to be examined
in methanol R and dilute to 5 m L with the same solvent.
Reference solution (a) Dissolve 20 mg o f bufexamac CRS in
methanol R and dilute to 10 m L with the same solvent.
Reference solution (b) Dissolve 10 m g o f salicylic acid R in
H I X "0 *"* and epimer at C* reference solution (a) and dilute to 5 m l. with the same
H solution.
H
Plate TLC silica gel F2 5 4 plate R.
Mobile phase glacial acetic acid R, dioxan R, toluene R
L. 16a, 17- [(1RS) -butylidenebis (oxy) ] -21 -hydroxypregna-1, 4- (4:20:90 V/V/V).
diene-3,11,20-trione. Application 10 jiL.
PhEur Development Over 2/3 o f the plate.
Drying In a current o f warm air.
Detection Examine in ultraviolet light at 254 nm .
System suitability, reference solution (b):
— the chromatogram shows 2 clearly separated spots.
Results T he principal spot in the chromatogram obtained w ith
the test solution is similar in position and size to the principal
spot in the chromatogram obtained w ith reference
solution (a).
TESTS
Related substances
Liquid chromatography (2.2.29).
1-332 Buflomedil Hydrochloride 2016
B. methyl 2-(4-butoxyphenyl)acetate,
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0 - 10 100 0
10 - 13 100-> 70 0-> 30
1 3-40 70 30
C. butyl 2-(4-butoxyphenyl)acetate,
Flow rate 1 m IJm in .
Detection Spectrophotom eter at 275 nm.
Injection 20 |iL.
Identification of impurities Use the chromatogram obtained
with reference solution (b) to identify the peaks due to
impurities A, B, C and D. D . 2-(4-butoxyphenyl)acetamide.
___________________________________________________________________________________ ' P h E ir
Relative retention W ith reference to bufexamac (retention
time = about 5.7 min): impurity D = about 1.3;
impurity A = about 1.8; impurity B = about 3.0;
impurity C = about 5.4.
System suitability. reference solution (b): Buflomedil Hydrochloride f**%
— resolution: minim um 2.0 between the peaks due to
bufexamac and impurity D . (Ph. Eux. monograph 1398) *
Limits:
— impurities A , B, C, D: for each impurity, not more than
the area o f the principal peak in the chromatogram
obtained with reference solution (a) (0.2 per cent);
— unspecified impurities: for each impurity, not more than
0.5 times the area o f the principal peak in the
chrom atogram obtained with reference solution (a)
C 17H 26d N 0 4 343.9 35543-24-9
(0.10 per cent);
— total: not more than 2.5 times the area of the principal
A c tio n a n d u se
peak in the chromatogram obtained with reference
Vasodilator.
solution (a) (0.5 p e r cent);
— disregard limit. 0.25 times the area o f the principal peak in P h & x ________________________________________________________________________________________ —
the chromatogram obtained with reference solution (a)
D E F IN IT IO N
(0.05 per cent).
4-(Pyrrolidin-l-yl)-l-(2,4,6-trim ethoxyphenyl)butan-l-one
L oss o n d ry in g (2.2.32) hydrochloride.
M axim um 0.5 per cent, determined on 1.000 g by drying
C o n te n t
in vacuo at 80 °C for 3 h.
98.5 per cent to 101.5 per cent (dried substance).
S u lfa te d a s h (2.4.14)
M axim um 0.1 per cent, determined on 1.0 g. CHARACTERS
A p p e a ra n c e
W hite or almost white, microcrystalline powder.
2016 Buflomedil Hydrochloride 1-333
OCH3 O r" \
TESTS
J Z K ^ O Appearance of solution
T he solution is clear (2.2.1) and colourless (2.2.2,
H O -^ ^ ^ O C H s Method II).
Dissolve 0.1 g in a 6 g/L solution o f potassium hydroxide R
B. 1-(4-hydroxy-2 j 6-dimethoxyphenyl)-4- (pyrrolidin - 1-
and dilute to 20 m L with the same solution.
yl)butan-l-one,
Related substances
och3 o r^ \ Liquid chromatography (2.2.29).
Test solution Dissolve 50 m g o f the substance to be examined
in the mobile phase and dilute to 25.0 m L with the mobile
HO ^ OH phase.
Reference solution (a) Dilute 1.0 m L o f the test solution to
C. 1-(2j4-dihydroxy-6-methoxyphenyl)-4-(pyrrolidin - 1- 100.0 m L with the mobile phase. Dilute 1.0 m L o f this
yl)butan-l-one. solution to 10.0 m L with the mobile phase.
PhEur Reference solution (b) Dissolve 2 m g o f bumetanide
impurity A CRS and 2 m g o f bumetanide impurity B CRS in
the mobile phase and dilute to 10.0 m L with the mobile
phase. Dilute 1.0 m L o f this solution to 100.0 m L with the
mobile phase.
Bumetanide ★ ★ Column:
***** — size: I = 0.15 m , 0 = 4.6 mm,
(Ph. Eur. monograph 1076)
— stationary phase: end-capped octylsUyl silica gelfor
chromatography R (3.5 fim).
C02H Mobile phase Mix 70 volumes o f methanol R, 25 volumes of
H,N
water for chromatography R and 5 volumes of a 27.2 g/L
solution o f potassium dihydrogen phosphate R previously
adjusted to p H 7.0 with a 280 g/L solution o f potassium
hydroxide R ,’ add tetrahexylammonium bromide R to this
mixture to obtain a concentration of 2.17 g/L.
Flow raze 1.0 mL/min.
C 1 7 H 2 0 N 2 O 5S 364.4 28395-03-1 Detection Spectrophotometer at 254 nm .
A c tio n a n d u se Injection 10 (iL.
Loop diuretic. Run time 5 times the retention time of bum etanide.
P re p a r a tio n s Relative retention W ith reference to bum etanide (retention
Bum etanide Injection time = about 6 min): impurity B = about 0.4;
Bum etanide Oral Solution impurity A = about 0.6; Impurity C = about 2.5;
impurity D = about 4.4.
Bumetanide Tablets
System suitability: reference solution (b):
Bum etanide and Slow Potassium Tablets — resolution: minimum 2.0 between the peaks due to
PhEur.
impurity A and im purity B.
L im its:
D E F IN IT IO N — impurities A, B, C, D: for each impurity, n o t m ore than
3-(Butylamino)-4-phenoxy-5-sulfamoylbenzoic acid. the area o f the principal peak in the chrom atogram
C o n te n t obtained with reference solution (a) (0.1 p er cent),
99.0 per cent to 101.0 per cent (dried substance). — any other impurity: for each impurity, n o t m ore th an the
CHARACTERS area o f the principal peak in the chrom atogram obtained
with reference solution (a) (0.1 per cent),
A p p e a ra n c e
— total: n o t more than twice the area of the principal peak in
W hite or alm ost white, crystalline powder.
the chromatogram obtained with reference solution (a)
S o lu b ility (0.2 per cent),
Practically insoluble in water, soluble in acetone and in — disregard limit: 0.5 times the area of the principal peak in
ethanol (96 p er cent), slightly soluble in methylene chloride. the chromatogram obtained with reference solution (a)
It dissolves in dilute solutions of alkali hydroxides. (0.05 per cent).
It shows polymorphism (5.9).
Loss on drying (2.2.32)
mp M aximum 0.5 per cent, determined on 1.000 g by drying in
A bout 233 °C. an oven at 105 °C for 4 h.
ID E N T IF IC A T IO N Sulfated ash (2.4.14)
Infrared absorption spectrophotometry (2.2.24). M aximum 0.1 per cent, determined on 1.0 g.
Comparison bumetanide CRS. ASSAY
If the spectra obtained in the solid state show differences, Dissolve 0.300 g in 50 m L o f alcohol R. Add 0.1 m L o f
dissolve the substance to be examined and the reference phenol red solution R. T itrate with 0.1 M sodium hydroxide
substance separately in acetone R, evaporate to dryness and until a violet-red colour is obtained. Carry out a blank
record new.spectra using the residues. titration.
2016 Bupivacaine Hydrochloride 1-335
CH3
0 O
*'/
.COoH
1 V ^ V
C 18H 29CIN 20 ,H 20 342.9 73360-54-0
>v nh2
Bupivacaine and Fentanyl Injection
PhEur.
DEFINITION
(2RS)-l -Butyl-N-(2 j6-dimethylphenyI)piperidine-2-
carboxamide hydrochloride monohydrate.
B. 3-amino-4-phenoxy-5-sulfamoylbenzoic acid. Content
98.5 per cent to 101.0 p e r cent (dried substance).
CHARACTERS
Appearance
White or almost white, crystalline powder or colourless
crystals.
Solubility
Soluble in water, freely soluble in ethanol (96 per cent).
IDENTIFICATION
C. butyl 3-(butylamino)-4-phenoxy-5-sulfamoylbenzoate, First identification A , D, E
Second identification B, C, D, E
A. Infrared absorption spectrophotometry (2.2.24).
Comparison bupivacaine hydrochloride CRS.
B. Thin-layer chromatography (2.2.27).
and enantiomer
Test solution Dissolve 25 mg of the substance to be examined
in methanol R and dilute to 5 m L with the same solvent.
Reference solution Dissolve 25 mg of bupivacaine
hydrochloride CRS in methanol R and dilute to 5 m L with the
D . 3- [ [(2ftS)-2-ethylhexyl] amino] -4-phenoxy-5- same solvent.
sulfamoylbenzoic acid. Plate TLC silica gel G plate R.
PhEur Mobile phase concentrated ammonia R, methanol R
(0.1:100 VIV).
Application 5 pL.
Development Over a path of 10 cm.
Drying In. air.
Detection Spray with dilute potassium iodobismuthate solution R.
Results T h e principal spot in the chromatogram obtained with
the test solution is similar in position, colour and size to the
principal spot in the chromatogram obtained with the
reference solution.
C. Dissolve 0.1 g in 10 m L of water R, add 2 m L of dilute
sodium hydroxide solution R and shake with 2 quantities, each
of 15 mL, o f 1,1-dimethylethyl methyl ether R. Dry the
combined upper layers over anhydrous sodium sulfate R and
filter. Evaporate the filtrate, recrystallise the residue from
1-336 Bupivacaine Hydrochloride 2016
ethanol (90 per cent VIV) R and dry under reduced pressure. Injection 1 jxL.
T he crystals m elt (2.2.14) at 105 °C to 108 °C. Identification of impurities Use the chrom atogram obtained
D. It gives reaction (a) o f chlorides (2.3.1). w ith reference solution (a) to identify the peaks due to
E. Optical rotation (see Tests). impurities B and E.
15 - 25 80 20
A. N- (2,6-dimethylphenyl)pyridine-2-carboxamide,
Flow rate 1.0 m L/m in.
Detection Spectrophotom eter at 240 nm.
Injection 50 |iL. and enantiomer
Identification of impurities Use the chromatogram obtained
with reference solution (a) to identify the peak due to
impurity F.
B. (2R5)-iV-(2,6-dimethylphenyl)piperidine-2-carboxamide,
Relative retention w ith reference to bupivacaine (retention
time = about 20 min): impurity F = about 0.3; methyl
benzoate = about 0.4.
System suitability:
— resolution: m inim um 4.0 between the peaks due to
impurity F and methyl benzoate in the chromatogram
obtained w ith reference solution (b);
— signal-to-noise ratio: minimum 40 for the principal peak in C . 1-(2,6-dimethylphenyl)-l,5,6,7-tetrahydro-2/i'-azepin-2-
the chrom atogram obtained with reference solution (a). one,
Limit.
— impurity F: n o t more than die area of the principal peak
in the chrom atogram obtained with reference solution (a) Cl and enantiomer
(10 ppm ).
H eavy m e ta ls ( 2.4.8)
Maximum 10 ppm .
D . (2ÆS)-2,6-dichloro-N-(2,6-dimethylphenyl)hexanamide,
Dissolve 2.0 g in a m ixture of 15 volumes of water R and
85 volumes o f methanol R and dilute to 20 m L with the same CH3
mixture o f solvents. 12 m L of the solution complies with
test B. Prepare the reference solution using lead standard ¿ X " r—
solution (1 p p m Pb) obtained by diluting lead standard
CH3 and enantiomer
solution (100 ppm Pb) R with a mixture o f 15 volumes of
water R and 85 volumes o f methanol R.
E. 6-(butylamino)-N-(2,6-dimethylphenyl)hexanamide,
L oss o n d ry in g (2.2.32)
4.5 per cent to 6.0 per cent, determined on 1.000 g by ch 3
drying in an oven at 105 °C.
S ulfa te d a sh (2.4.14)
M aximum 0.1 p er cent, determined on 1.0 g. CH3
A SSA Y
Dissolve 0.250 g in a mixture of 20 m L o f water R and F . 2,6-dimethylaniline.
25 m L o f ethanol (96per cent) R. Add 5.0 m L o f 0.01 M PhEur
hydrochloric add. C arry out a potentiometric titration (2.2.20),
using 0.1 M ethanolic sodium hydroxide. Read the volume
added betw een the 2 points of inflexion.
1 m L o f 0.1 M ethanolic sodium hydroxide is equivalent to ★ *
32.49 m g o f C 18H 29CIN 2O.
Buprenorphine ★ ★
(Ph. Eur. monograph 1180) *****
STO RA G E
Protected from light.
IM P U R IT IE S
Specified impurities B, F
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the m onograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general m onograph Substances for pharmaceutical use C 29H 41NO 4 52485-79-7
(2034). It is therefore not necessary to identify these
impurities for dem onstration o f compliance. See also 5.10. A c tio n a n d u se
Opioid receptor partial agonist; analgesic.
1-338 Buprenorphine 2016
12- 15 64 -» 41 36 -» 59
15 - 20 41 -> 39 59 -» 61
D . R I = R2 = C H 3: ( 2.S)-2-[l 7-(cydopropylmethyl)-4J5a-
epoxy-3,6-dimethoxy-6a, 14-ethano-14a-m orphinan-7 a-yl] -
C 29H 42CINO 4 504.1 53152-21-9
3j3-dimethylbutan-2-ol (3-O-methylbuprenorphine),
E. R I = R2 = H : (25)-2-[17-(cyclopropylmethyl)-4,5a- A ctio n a n d u se
epoxy-3j6-dihydroxy-6a, 14-ethano-14oc-morphinan-7 a-yl] - Opioid receptor partial agonist; analgesic.
3,3-dimethylbutan-2-ol (6-O-desmethylbuprenorphine),
P re p a r a tio n s
Buprenorphine Injection
Buprenorphine Transdermal Patches
Buprenorphine Sublingual Tablets
PhEur__________________________________________________________
D E F IN IT IO N
(2S)-2-[17-(Cyclopropylmethyl)-4,5a-epoxy-3-hydroxy-6-
methoxy- 6a , 14-ethano-14a-m orphinan-7 a-yl] -3,3-
F. 17-(cyclopropyimethyl)-4j5a-epoxy-6-methoxy-7a-[l-(l j 1
dimethylbutan- 2-ol hydrochloride.
dimethylethyi)ethenyl] - 6a, 14-ethano-14a-m orphinan-3-ol,
C o n te n t
98.5 per cent to 101.5 p er cent (dried substance).
CHARACTERS
A p p e a ra n c e
W hite or almost white, crystalline powder.
S o lu b ility
Sparingly soluble in water, freely soluble in methanol, soluble
in ethanol (96 per cent), practically insoluble in cyclohexane.
G. R-R; 17,17 '-diCcyclopropyimethyO^jSa^' jSa'-diepoxy-
ID E N T IF IC A T IO N
7 a ,7 a /-di[(lS)-l-hydroxy-l,2,2-trim ethylpropyl]-6,6/-
A. Infrared absorption spectrophotometry (2.2.24).
dimethoxy-2,2 '-bi( 6a , 14-ethano-14a-m orphinan)-3,3 '-diol
(2,2 '-bibuprenorphine), Comparison buprenorpkine hydrochloride CRS.
B. 3 m L o f solution S (see Tests) gives reaction (a) of
chlorides (2.3.1).
1-340 Buprenorphine Hydrochloride 2016
TESTS Limits:
Solution S — correction factor, for the calculation of content, multiply the
Dissolve 0.250 g in 5.0 m L of methanol R and, while stirring, peak area of impurity G by 0.3;
dilute to 25.0 m L with carbon dioxide-free water R. — impurity H: not more than 2.5 times the area of the
Appearance o f solution principal peak in the chromatogram obtained with
Solution S is clear {2.2.1) and colourless (2.2.2, Method II). reference solution (a) (0.25 per cent);
— impurities A, B, F, J: for each impurity, not more than
A cid ity o r alk a lin ity twice the area of the principal peak in the chromatogram
T o 10.0 m L of solution S add 0.05 m L of methyl red obtained with reference solution (a) (0.2 per cent);
solution R. N o t m ore than 0.2 m L o f 0.02 M sodium hydroxide — impurity G: not m ore than 1.5 times the area o f the
or 0.02 M hydrochloric acid is required to change the colour of principal peak in the chromatogram obtained with
the indicator. reference solution (a) (0.15 per cent);
Specific optical rotation (2.2.7) — unspecified impurities: for each impurity, n o t m ore than the
—92 to - 9 8 (dried substance). area of the principal peak in the chromatogram obtained
Dissolve 0.200 g in methanol R and dilute to 20.0 m L with with reference solution (a) (0.10 per cent);
the same solvent. — total: not more than 7 times the area o f the principal peak
in the chromatogram obtained with reference solution (a)
R e la te d su b s ta n c e s
(0.7 per cent);
Liquid chromatography (2.2.29).
— disregard limit: 0.5 times the area o f the principal peak in
Test solution Dissolve 50.0 m g of the substance to be the chromatogram obtained with reference solution (a)
examined in methanol R and dilute to 10.0 m L with the same (0.05 per cent).
solvent.
L o ss on d ry in g (2.2.32)
Reference solution (a) Dilute 1.0 m L o f the test solution to
M aximum 1.0 per cent, determ ined on 1.000 g by heating in
100.0 m L with methanol R. Dilute 1.0 m L of this solution to
an oven at 115-120 °C.
10.0 m L with methanol R.
Reference solution (b) Dissolve 5 m g of buprenorphine for system ASSA Y
suitability CRS (containing impurities A, B, F, G, H and J) in Dissolve 0.400 g in a mixture o f 5 m L of 0.01 M hydrochloric
1.0 m l, of methanol R. acid and 50 m L o f ethanol (96 per cent) R. Carry out a
potentiometric titration (2.2.20), using 0.1 M sodium
Column:
— size. I = 0.05 m , 0 = 4.6 mm; hydroxide. Read the volume added between the 2 points of
inflexion. Carry out a blank titration.
— stationary phase: end-capped octadecylsUyl silica gel for
chromatography R (3.5 pm); 1 m L o f 0.1 M sodium hydroxide is equivalent to 50.41 mg of
— temperature. 30 °C. C29H42CINO4.
Mobile phase: STO R A G E
— mobile phase A: mix 10 volumes of acetonitnle R and Protected from light.
90 volumes of the following solution: dissolve 5.44 g of
IM P U R IT IE S
potassium dihydrogen phosphate R in 900 m L o f water R,
adjust to p H 4.5 with a 5 per cent V/V solution of Specified impurities A , B, F, G, H, J.
phosphoric acid R and dilute to 1000 m L with water R', Other detectable impurities (the following substances would, if
— mobile phase B: acetonitnle R; present at a sufficient level, be detected by one or other of
the tests in the m onograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
Time Mobile phase A Mobile phase B
by the general monograph Substances for pharmaceutical use
(min) (per cent V/V) (per cent V/V)
(2034). It is therefore not necessary to identify these
0- 2 89 11
impurities for demonstration of compliance. See also 5.10.
2 - 12 89 -»64 11 -» 36 Control of impurities in substances for pharmaceutical use): C, D,
12 - 15 64 -»41 36-» 59 E, I.
15 - 20 41 -» 39 59-» 61
,R
N
Buserelin *****
★ ★
(Ph Ever monograph 1077) *
C. 4,5a-epoxy-7a-[(l.S)-l-hydroxy-l,2,2-triinethylpropyl]-
3, 6-dimethoxy-6a, 14-ethano-14a-m orphinan-17-carbonitrile, H j|-His-Trp-Ser-Tyr-D-Ser-Leu-Arg-Pro-N CH3
O
Q oH m N wO u 1239 57982-77-1
A ctio n a n d u se
Gonadotrophin releasing hormone (gonadorelin) analogue;
treatm ent of prostate cancer.
0.9 to 1.1. N o t more than traces of other amino acids are Test solution Dissolve 20.0 mg of the substance to be
present. examined in a mixture o f 5 volumes of mobile phase B and
95 volumes of mobile phase A and dilute to 10.0 m L with
TESTS
the same mixture of solvents.
A p p e a ra n c e o f so lu tio n
A 10 g/L solution is clear (2.2.1) and not more intensely W a te r (2.5.12)
coloured than reference solution Y 7 (2.2.2, Method IT). M aximum 4.0 per cent, determined on 80.0 mg.
S pecific o p tic a l ro ta tio n (2.2.7) B a c te ria l e n d o to x in s (2.6.14)
- 4 9 to - 5 8 (anhydrous, acetic acid-free substance), Less than 55.5 IU/mg, if intended for use in the manufacture
determined on a 10 g/L solution. of parenteral preparations without a further appropriate
procedure for the removal o f bacterial endotoxins.
Specific a b so rb a n c e (2.2.25)
49 to 56, m easured at the absorption maximum at 278 nm A SSAY
(anhydrous, acetic acid-free substance). Liquid chromatography (2.2.29) as described in the test for
Dissolve 10.0 mg in 100.0 m L of 0.01 M hydrochloric add. related substances with the following modification.
R e la te d su b s ta n c e s Injection T est solution and reference solution (b).
Liquid chromatography (2.2.29). Calculate the content o f buserelin ( C ^ H s ^ ^ O ^ ) using the
Test solution Dissolve 5.0 mg of the substance to be examined areas o f the peaks in the chromatograms obtained and the
in 5.0 m L of the mobile phase. declared content of C 60H 86N 16O i 3 in buserdin CRS.
Reference solution (a) Dissolve the contents o f a vial of D-His- STO R A G E
buserdin CRS in the mobile phase. Dilute an In an airtight container, protected from light, at a
appropriate volume o f this solution in the mobile phase to tem perature o f 2 °C to 8 °C. If the substance is sterile, store
obtain a final concentration o f 1 mg/mL. Add 1.0 m L o f the in an airtight, sterile, tam per-proof container.
test solution to 1.0 m L o f this solution.
L A B E L L IN G
Reference solution (b) Dissolve the contents of a vial of T h e label states:
buserdin CRS in the mobile phase. Dilute an — the mass of peptide in the container;
appropriate volume o f this solution in the mobile phase to — where applicable, that the substance is suitable for use in
obtain a final concentration o f 1.0 mg/mL. the manufacture o f parenteral preparations.
Reference solution (c) Dilute 1.0 m L of the test solution to
IM P U R IT IE S
100.0 m L with the mobile phase.
Spedfied impurities: A, B, C, D , E.
Column:
— size: I = 0.25 m , 0 = 4 mm ;
— stationary phase: octadecylsilyl silica gd for chromatography R
(5 |un).
H
Mobile phase Mix 200 m L of acetomtrUe R and 700 m L o f an
11.2 g/L solution o f phosphoric add R and adjust to p H 2.5 H jp D-His - Trp - Ser - Tyr - D-Ser - Leu - Arg - Pro - N CH3
with triethylamine R. O
Flow rate 0.8 m U m in.
A. [2-D-histidine]buserelin,
Detection Spectrophotom eter at 220 nm .
Injection 10 pL of the test solution, reference solution (a) and
reference solution (c).
Relative retention W ith reference to buserelin (retention
time = about 36 m in): impurity B = about 0.76;
H |j—His - Trp - D-Ser - Tyr - D-Ser - Leu - Arg - Pro - N CH 3
impurity C = about 0.83; impurity A = about 0.90;
impurity D = about 0.94; impurity E = about 0.94. O
System suitability: reference solution (a):
— resolution: m inim um 1.5 between the peaks due to B. [4-D-serine]buserelin,
impurity A and buserelin.
Limits: H3C ^ H3
— sum of impurities D and E: not more than 3 times the area
o f the principal peak in the chromatogram obtained with H-Trp-Ser-Tyr-D-Ser-Leu-Arg-Pro-N CH3
H
reference solution (c) (3 per cent);
— arty other impurity: for each impurity, not more than
3 times the area o f the principal peak in the C. buserelin-(3-9)-peptide,
chrom atogram obtained with reference solution (c)
(3 per cent);
— total: not more than 5 times the area of the principal peak
in the chrom atogram obtained with reference solution (c)
(5 per cent); H His - Trp - Ser-D -Tyr-D-Ser-Leu-A rg-Pro-N CH3
— disregard limic. 0.1 times the area o f the principal peak in O
the chrom atogram obtained with reference solution (c)
(0.1 per cent).
D . [5-D-tyrosine]buserelin,
A cetic a c id (2.5.34)
3.0 per cent to 7.0 per cent.
2016 Buspirone Hydrochloride 1-343
Column:
ch 3 — size. I = 0.15 m , 0 = 4.6 mm,
Ç t H3C
p"CH3 — stationary phase: octadecylsHyl silica gelfor chromatography R
oI
H V -H is- Trp-Ser-Tyr-D-Ser-Leu-Arg-Pro-N CH3 (5 pm),
H
O — temperature: 40 °C.
Mobile phase:
E. [l-(5-oxo-D-proline)]buserelin. — mobile phase A: mix 950 volumes of a solution containing
PhEur 6.8 g/L of potassium dihydrogen phosphate R and 0.93 g/L
of sodium hexanesulfonate monohydrate R, previously
adjusted to p H 3.4 with phosphoric acid R and 50 volumes
of acetorntrik Rl;
>** ★ — mobile phase B: mix 250 volumes of a solution containing
★
Buspirone Hydrochloride ★ * 3.4 g/L of potassium dihydrogen phosphate R and 3.52 g/L
***** of sodium hexanesulfonate monohydrate R, previously
(Ph Eur monograph 1711) adjusted to p H 2.2 with phosphoric acid R and
750 volumes of acetorntrik R l,
D E F IN IT IO N
Flow rate 1 mL/min.
8- [4- [4- (Pyrimidin-2-yl) piperazin-1-yl] butyl] -8 -
azaspiro[4.5]decane-7,9-dione hydrochloride. Detection Variable wavelength spectrophotometer capable of
operating at 240 nm and at 210 run.
Content
99.0 per cent to 101.0 per cent (dried substance).
Injection 20 |oL.
Identification of impurities Use the chromatogram supplied
CHARACTERS with buspirone for system suitability CRS and the
Appearance chromatogram obtained with reference solution (b) to
White or alm ost white, crystalline powder. identify the peaks due to impurities E, G, J, L and N .
Solubility Relative retention at 240 nm With reference to buspirone
Freely soluble in water and in methanol, practically insoluble (retention time = about 25 min): impurity A = about 0.2;
in acetone. impurity B = about 0.3; impurity C = about 0.6;
It shows polymorphism (5.9). impurity D = about 0.7; impurity E = about 0.8;
IDENTIFICATION impurity F = about 0.9; impurity G = about 1.05;
impurity H = about 1.1; impurity I = about 1.2;
A. Infrared absorption spectrophotometry (2.2.24).
im purity J = about 1.5.
Comparison buspirone hydrochloride CRS.
Relative retention at 210 nm With reference to buspirone
If the spectra obtained in the solid state show differences, (retention time = about 25 min): impurity K = about 0.6;
dissolve the substance to be examined and the reference impurity L = about 1.7; impurity M = about 1.8;
substance separately in methanol R, evaporate to dryness on a impurity N = about 1.9.
water-bath and record new spectra using the residues.
System suitability: reference solution (b):
B. It gives reaction (a) of chlorides (2.3.1). — peak-to-valley ratio at 240 nm: m in im u m 5.0, where
TESTS Hp = height above the baseline of the peak due to
Related substances impurity G and Hv = height above the baseline of the
Liquid chromatography (2.2.29). lowest point o f the curve separating this peak from the
peak due to buspirone;
Test solution Dissolve 25.0 mg of the substance to be
examined in mobile phase A and dilute to 25.0 m L with — resolution at 210 nm: minimum 4.0 between the peaks due
mobile phase A. to impurity L and impurity N ;
— the chromatograms obtained are similar to the
Reference solution (a) Dilute 1.0 mL o f the test solution to
chromatograms supplied with buspirone for system
100.0 m L with mobile phase A. Dilute 1.0 m L of this
suitability CRS.
solution to 10.0 m L with mobile phase A.
Limits Spectrophotometer at 240 nm :
Reference solution (b) Dissolve the contents o f a vial of — correction factor, for the calculation of contentj multiply the
buspirone for system suitability CRS (containing impurities E, peak area of impurity J by 2,
G, J 3L and N ) in 2.0 ml of mobile phase A and sonicate for
10 min.
1-344 Buspirone Hydrochloride 2016
L oss o n d ry in g (2.2.32)
M axim um 0.5 per cent, determined on 1.000 g by drying at
105 °C.
S u lfa te d a s h (’2.4.14)
M aximum 0.1 per cent, determined on 1.0 g.
A SSAY
Dissolve 0.150 g in 10 m L o f glacial acetic add R and add
50 m L o f acetic anhydride R. T itrate with 0.1 M perchloric
acid, determining the end-point potentiometrically (2.2.20).
1 m L of 0.1 M perchloric add is equivalent to 21.10 mg F. X = N H : 4-[4-(pyrimidin-2-yl)piperazin-l-yl]butyl [l-[ 2 -
o f C 2JH 32CIN 5O 2. oxo-2- [ [4- [4-(pyrimidin-2-yl)piperazin-1-
STORAGE yl] butyl] amino] ethyl] cyclopentyl] acetate,
Protected from light. H . X = O: bis[4-[4-(pyrimidin-2-yl)piperazin-l-yl]butyl]
(cyclopentane- 1,1 -diyl) diacetate,
IM P U R IT IE S
a
Spedfied impurities E, J, K
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other o f N N
the tests in the monograph. T hey are limited by die general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these
impurities for demonstration o f compliance. See also 5.10.
G. 2, 2 '-(piperazine- 1,4-diyl) dipyrimidine,
Control of impurities in substances for pharmaceutical use): A , B,
C, D, F, G, H, I, L, M , N.
■v
Y l
n ' n^ x ci
A. 2-(piperazm-l-yl)pyrimidine,
I. 8-[4-[4-(5-chloropyrimidin-2-yl)piperazin-l-yl]butyl]-8-
azaspiro [4.5] decane-7,9-dione,
2016 Busulfan 1-345
ID E N T IF IC A T IO N
First identification A.
Second identification B, C, D.
A. Infrared absorption spectrophotometry (2.2.24).
Comparison busulfan CRS.
B. Thin-layer chromatography (2.2.27).
Test solution Dissolve 20 mg of the substance to be examined
J. 4-(7,9-dioxo-8-azaspiro[4.5]dec-8-yl)butyl [l-[2-oxo-2-[[4- in 2 m L o f acetone R.
[4-(pyrimidin- 2-yl)piperazin - 1- Reference solution Dissolve 20 mg o f busulfan CRS in 2 m L of
yl] butyl] amino] ethyl] cyclopentyl] acetate. acetone R.
Plate TLC silica gel G plate R.
Mobile phase acetone R, toluene R (50:50 VIV).
Application 5 (¿L.
Y ' Development Over a p ath of 15 cm.
Drying In a current of warm air.
K. R = H: 8-a2aspiro [4.5] decane-7,9-dione, Detection Spray with anisaldehyde solution R and heat at
L. R = [CHJ4-CI: 8-(4-chlorobutyl)-8-azaspiro [4.5] decane- 120 °C.
7, 9-dione, Results T h e principal spot in the chromatogram obtained with
M . R — [C H ^ -B r: 8-(4-bromobutyl)-8-azaspiro[4.5]decane- the test solution is similar in position, colour and size to the
7,9-dione, principal spot in the chromatogram obtained with the
reference solution.
C. T o 0.1 g add 5 m L o f 1 M sodium hydroxide. H eat until a
clear solution is obtained. Allow to cool. T o 2 m L of the
solution add 0.1 m L o f potassium permanganate solution R.
T h e colour changes from purple through violet to blue and
finally to green. Filter and add 1 m L of ammoniacal silver
nitrate solution R. A precipitate is formed.
D . T o 0.1 g add 0.1 g o f potassium nitrate R and 0.25 g of
N . 8 ,8 '-(butane- 1,4-diyl)bis(8-azaspiro [4.5] decane-7,9-
sodium hydroxide R, mix and heat to fusion. Allow to cool and
dione).
dissolve the residue in 5 m L of water R. Adjust to pH 1-2
P h E ir using dilute hydrochloric acid R. T h e solution gives reaction (a)
of sulfates (2.3.1).
TESTS
A p p e a ra n c e o f so lu tio n
Busulfan ★ ★
★ ★ T h e solution is clear (2.2.1) and n o t more intensely coloured
*★★★* than reference solution B 7 (2.2.2, Method II).
(Ph. Eitr. monograph 0542)
Dissolve 0.25 g in 20 m L o f acetonitrile R, dilute to 25 m L
0\\ //0 with water R and examine immediately.
, 0 ^ ,c h 3 A cid ity
h3c 0 s
//* Dissolve 0.20 g with heating in 50 m L of anhydrous
o o
ethanol R. Add 0.1 m L of methyl red solution R. N o t more
Q H ^O ^ 246.3 55-98-1 than 0.05 m L o f 0.1 M sodium hydroxide is required to
change the colour of the indicator.
A c tio n a n d u se L oss o n d ry in g (2.2.32)
Cytotoxic alkylating agent. M aximum 2.0 per cent, determined on 1.000 g by drying
P re p a r a tio n in vacuo at 60 °C.
Busulfan Tablets S u lfate d a sh (2.4.14)
M aximum 0.1 p er cent, determined on 1.0 g.
P h E ir .
ASSAY
D E F IN IT IO N
T o 0.250 g add 50 m L o f water R. Shake. Boil under a reflux
B utane- 1,4-diyl di(methanesulfonate).
condenser for 30 min and, if necessary, make up to the
C o n te n t initial volume with water R. Allow to cool. Using 0.3 m L o f
99.0 per cent to 100.5 per cent (dried substance). phenolphthalein solution R as indicator, titrate with 0.1 M
CHARACTERS sodium hydroxide until a pink colour is obtained.
A p p e a ra n c e 1 m L of 0.1 M sodium hydroxide is equivalent to 12.32 mg of
W hite or almost white, crystalline powder. C sH uO ^.
S o lu b ility STORA GE
Very slightly soluble in water, freely soluble in acetone and in In an airtight container, protected from light.
acetonitrile, very slightly soluble in ethanol (96 per cent).
__________________________________________________________________________________________ P h E ir
mp
A bout 116 °C.
1-346 Butyl Hydroxybenzoate 2016
TESTS
Butyl Hydroxybenzoate ***** Solution S
Butylparaben * ** Dissolve 1.0 g in ethanol (96 per cent) R and dilute to
(Butyl Parahydroxybenzoate, Ph Eur monograph 0881) 10 m L with the same solvent.
Appearance o f solution
Solution S is clear (2.2.1) and n o t more intensely coloured
than reference solution BY 6 (2.2.2, Method II).
Acidity
T o 2 m L o f solution S add 3 m L o f ethanol (96 per cent) R,
5 m l. of carbon dioxide-free water R and 0.1 m l. of bromocresol
C n H 140 3 194.2 94-26-8 green solution R. N o t more than 0.1 m L o f 0.1 M sodium
hydroxide is required to change the colour o f the indicator to
Action and use blue.
Excipient. Related substances
Liquid chromatography (2.2.29).
Ph Eur'___________________________________________________________
Test solution Dissolve 50.0 m g o f the substance to be
D E F IN IT IO N examined in 2.5 m L of methanol R and dilute to 50.0 m l.
Butyl 4-hydroxybenzoate. with the mobile phase. D ilute 10.0 m L o f the solution to
Content 100.0 m L with the mobile phase.
98.0 per cent to 102.0 per cent. Reference solution (a) Dissolve 5 m g o f 4-hydroxybenzoic acid R
CHARACTERS (impurity A), 5 m g of propyl parahydroxybenzoate R
Appearance (impurity D ) and 5 mg o f the substance to be examined in
W hite or alm ost white, crystalline powder or colourless the mobile phase and dilute to 100.0 m L with the mobile
crystals. phase. Dilute 1.0 m L o f the solution to 10.0 m L with the
mobile phase.
Solubility
Reference solution (b) Dissolve 50.0 mg o f butyl
Very slightly soluble in water, freely soluble in ethanol
parahydroxybenzoate CRS in 2.5 m L of methanol R and dilute
(96 per cent) and in m ethanol.
to 50.0 m L with the mobile phase. D ilute 10.0 m L o f the
IDENTIFICATION solution to 100.0 m L with the mobile phase.
First identification A , B Reference solution (c) Dilute 1.0 m L o f the test solution to
Second identification A , C 20.0 m L with the mobile phase. D ilute 1.0 m L of this
A. Melting point (2.2.14): 68 °C to 71 °C. solution to 10.0 m L with the mobile phase.
B. Infrared absorption spectrophotom etry (2.2.24). Reference solution (d) Dissolve 5 m g of butyl
Comparison butyl parahydroxybenzoate CRS. parahydroxybenzoate impurùy E CRS (iso-butyl
parahydroxybenzoate) in the mobile phase and dilute to
C. Thin-layer chrom atography (2.2.27).
100.0 m L with the mobile phase.
Test solution (a) Dissolve 0.10 g of the substance to be
Reference solution (e) Dilute 0.5 m L o f reference solution (d)
examined in acetone R and dilute to 10 m L with the same
to 50.0 m L with reference solution (b).
solvent.
Column:
Test solution (b) D ilute 1 m L of test solution (a) to 10 m L — size. I = 0.15 m , 0 = 4.6 m m ;
with acetone R.
— stationary phase: end-capped octadecylsilyl silica gel for
Reference solution (a) Dissolve 10 mg o f butyl chromatography R (5 pm);
parahydroxybenzoate CRS in acetone R and dilute to 10 m L — temperature: 35 °C.
with the sam e solvent.
Mobile phase 6.8 g/L solution o f potassium dihydrogen
Reference solution (b) Dissolve 10 mg o f propyl phosphate R, methanol R (50:50 V/V).
parahydroxybenzoate R in 1 m L of test solution (a) and dilute Flow rate 1.3 mL/min.
to 10 m L w ith acetone R.
Detection Spectrophotom eter at 272 nm.
Plate TLC octadecylsilyl silica gel F2 5 4 plate R.
Injection 10 pL o f the test solution and reference
Mobile phase glacial acetic add R, water R, methanol R
solutions (a), (c) and (e).
(1:30:70 VIVIV).
Run time 1.5 times the retention time o f butyl
Application 2 pL of test solution (b) and reference
parahydroxybenzoate.
solutions (a) and (b).
Identification of impurities U se the chrom atogram obtained
Development Over 2/3 of the plate.
with reference solution (a) to identify the peaks due to
Drying In air. impurities A and D ; use the chrom atogram obtained with
Detection Examine in ultraviolet light at 254 nm. reference solution (e) to identify the peak due to im purity E.
System suitability: reference solution (b): Relative retention W ith reference to butyl parahydroxybenzoate
— the chrom atogram shows 2 clearly separated principal (retention time = about 22 min): impurity A = about 0.1;
spots. impurity D = about 0.5; im purity E = about 0.9.
Results T he principal spot in the chromatogram obtained with System suitability:
test solution (b) is similar in position and size to the principal — resolution:
spot in the chrom atogram obtained with reference
solution (a).
2016 Butylated Hydroxyanisole 1-347
R e la te d su b sta n c e s
Examine by thin-layer chromatography (2.2.27), using silica
Butylated Hydroxytoluene ?***
gel G R as the coating substance. *★ ★*
(Butylhydroxytoluene, Ph Eur monograph 0581) *
Test solution (a) Dissolve 0.25 g of the substance to be
examined in methylene chloride R and dilute to 10 m L with
the same solvent.
Test solution (b) D ilute 1 m L o f test solution (a) to 10 m L
with methylene chloride R.
Reference solution (a) Dissolve 25 mg o f
butylhydroxycmisole CRS in methylene chloride R and dilute to
10 m L with the same solvent.
C i 5H 240 220.4 128-37-0
Reference solution (b) D ilute 1 m L o f reference solution (a) to
20 m L with methylene chloride R. A c tio n a n d u se
Reference solution (c) Dissolve 50 m g o f hydroquinone R in Antioxidant.
5 m L of alcohol R and dilute to 100 m L with methylene
chloride R. Dilute 1 m L of this solution to 10 m L with P h E i r ____________________________________________________________________________________________
methylene chloride R. D E F IN IT IO N
Apply separately to the plate 5 jiL of each solution. Develop Butylhydroxytoluene is 2,6-bis (1,1 -dimethylethyl)-4-
over a path o f 10 cm using methylene chloride R. Allow the m ethylphenol.
plate to dry in air and spray with a freshly prepared mixture
CHARACTERS
of 10 volumes of potassium ferricyanide solution R, 20 volumes
A white or yellowish-white, crystalline powder, practically
of ferric chloride solution R1 and 70 volumes of water R. In the
insoluble in water, very soluble in acetone, freely soluble in
chrom atogram obtained with test solution (a): any violet-blue
alcohol and in vegetable oils.
spot with an RF value o f about 0.35 (corresponding to
3-(l,l-dim ethylethyl)-4-m ethoxyphenol) is not m ore intense ID E N T IF IC A T IO N
than the principal spot in the chrom atogram obtained with First identification A , C.
reference solution (a) (10 per cent); any spot corresponding Second identification A , B, D.
to hydroquinone is n o t more intense than the principal spot
A. Freezing-point (see Tests).
in the chromatogram obtained with reference solution (c)
B. Dissolve 0.500 g in ethanol R and dilute to 100.0 m L with
(0.2 per cent); any spot, apart from the principal spot and
the same solvent. Dilute 1.0 m L o f this solution to 100.0 m L
any spots corresponding to 3-(l,l-dim ethylethyl)-4-
w ith ethanol R. Examined between 230 n m and 300 nm
methoxyphenol and hydroquinone, is n o t more intense than
the principal spot in the chrom atogram obtained with
(2.2.25), the solution shows an absorption m aximum at
278 nm . T h e specific absorbance at the m aximum is 80 to
reference solution (b) (0.5 p er cent).
90.
H eav y m e ta ls (2.4.8)
C . Examine by infrared absorption spectrophotom etry
1.0 g complies with test C for heavy metals (10 ppm).
(2.2.24), com paring with the spectrum obtained with
Prepare the reference solution using 1 m L o f lead standard
butylhydroxytoluene CRS.
solution (10 ppm Pb) R.
D . Dissolve about 10 mg in 2 m L o f alcohol R. Add 1 m L o f
S u lfa te d a s h (2.4.14)
a 1 g/L solution o f testosterone propionate R in alcohol R and
N ot more than 0.1 per cent, determ ined on 1.0 g.
2 m L o f dilute sodium hydroxide solution R. H eat in a water-
STORAGE b ath at 80 °C for 10 min and allow to cool. A blue colour
Store protected from light. develops.
IM P U R IT IE S TESTS
A p p e a ra n c e o f so lu tio n
Dissolve 1.0 g in methanol R and dilute to 10 m L with the
same solvent. T h e solution is clear (2.2.1) and not m ore
intensely coloured than reference solution Y 5 or BY 5 (2.2.2,
Method II).
A. benzene- 1,4-diol (hydroquinone). F re e z in g -p o in t (2.2.18)
____________________________________________________________________________________________PhEur 69 °C to 70 °C.
R e la te d su b s ta n c e s
Exam ine by thin-layer chromatography (2.2.27), using silica
gel G R as th e coating substance.
Test solution Dissolve 0.2 g o f the substance to be examined
in methanol R and dilute to 10.0 m L with the same solvent.
Reference solution Dilute 1 m L o f the test solution to 200 m L
w ith methanol R.
Apply separately to the plate 10 |iL o f each solution. Develop
over a p ath o f 15 cm using methylene chloride R. D ry the plate
in air and spray with a freshly prepared mixture of
10 volumes o f potassium ferricyanide solution R, 20 volumes o f
ferric chloride solution R1 and 70 volumes o f water R. Any spot
in the chrom atogram obtained with the test solution, apart
2016 Cabergoline 1-349
from the principal spot, is not more intense than the spot in Reference solution (a) Dissolve 30.0 mg of cabergoline CRS in
the chromatogram obtained with the reference solution the mobile phase and dilute to 25.0 m L with the mobile
(0.5 per cent). phase.
S u lfa te d a s h (2.4.14) Reference solution (b) Dilute 1.0 m L of the test solution to
N ot m ore than 0.1 per cent, determined on 1.0 g. 100.0 m L with the mobile phase. Dilute 10.0 m L of this
Ph Eur
solution to 50.0 m L with the mobile phase.
Reference solution (c) Suspend 50 m g of the substance to be
examined in 10 m L of 0.1 M sodium hydroxide. Stir for about
15 min. T o 1 m L o f the suspension add 1 m L o f 0.1 M
.**★ ★ hydrochloric acid and dilute to 10 m L with the mobile phase.
★
Cabergoline ★ ★ Sonicate until dissolution is complete. T h e main degradation
***** product obtained is impurity A.
(Ph. Eur. monograph 1773)
Column'.
— size. I = 0.25 m , 0 = 4.6 mm,
— stationary phase. octadecylsQyl silica gel for chromatography R
(10 nm).
Mobile phase Mix 16 volumes of acetonitrUe R and 84 volumes
o f a freshly prepared 6.8 g/L solution of potassium dihydrogen
phosphate R previously adjusted to p H 2.0 with phosphoric
H H « 11
0 0 add R. Add 0.2 volumes of triethylamine R.
Flow rate 1.2 mL/min.
Detection Spectrophotometer at 280 nm.
C 26H 37N 5O 2 451.6 81409-90-7
Injection 20 |iL of the test solution and reference solutions (b)
A ctio n a n d u se and (c).
Dopam ine D 2 receptor agonist. Run time 4 times the retention time o f cabergoline.
PhEur___________________________
Relative retention W ith reference to cabergoline (retention
time = about 12 min): impurity D = about 0.3;
D E F IN IT IO N impurity B = about 0.6; impurity A = about 0.8;
1-Ethyl-3- [3-(dimethylamino)propyl] -3-[[(6ai?,9i?, 10ai?)-7- impurity C = about 2.9.
(prop-2-enyl)-4,6,6a,7,8,9,10,1 Oa-octahydroindolo [4,3- System suitability', reference solution (c):
j£]quinolin-9-yl] carbonyl]urea. — resolution: minimum 3.0 between the peaks due to
C o n te n t cabergoline and impurity A.
98.0 per cent to 102.0 per cent (anhydrous substance). Limits:
CHARACTERS — impurities A, C: for each impurity, not more than
A p p e a ra n c e 1.5 times the area of the principal peak in the
White or almost white, crystalline powder. chromatogram obtained with reference solution (b)
(0.3 per cent);
S o lu b ility
— impurities B, D: for each impurity, not more than
Practically insoluble in water, freely soluble in ethanol
0.5 times the area of the principal peak in the
(96 per cent), very slightly soluble in hexane. It is slightly
chromatogram obtained with reference solution (b)
soluble in 0.1 M hydrochloric add.
(0.1 per cent);
It shows polymorphism (5.9). — any other impurity, for each impurity, n o t more than
ID E N T IF IC A T IO N 0.5 times the area of the principal peak in the
A. Specific optical rotation (see Tests). chromatogram obtained with reference solution (b)
(0.1 per cent);
B. Infrared absorption spectrophotometry (2.2.24).
— total: not more than 4 times the area o f the principal peak
Comparison cabergoline CRS. in the chromatogram obtained with reference solution (b)
If the spectra obtained in the solid state show differences, (0.8 per cent);
dissolve 50 mg o f the substance to be examined and 50 mg — disregard limit. 0.25 times the area o f the principal peak in
o f the reference substance separately in 1 m L o f ethanol the chromatogram obtained with reference solution (b)
(96 per cent) R, evaporate to dryness and record new spectra (0.05 per cent).
using the residues.
W a te r (2.5.12)
TESTS M aximum 0.5 per cent, determined on 1.000 g.
S pecific o p tic a l ro ta tio n (2.2.7) A SSAY
—77 to —83 (anhydrous substance).
Liquid chromatography (2.2.29) as described in the test for
Dissolve 0.100 g in ethanol (96 per cent) R and dilute to related substances with the following, modification.
50.0 m L with the same solvent.
Injection T est solution and reference solution (a).
R e la te d su b sta n c e s Calculate the percentage content of C 26H 37N 5O 2 from the
Liquid chromatography (2.2.29). Prepare the solutions areas o f the peaks and the declared content of
immediately before use and protectedfrom light. cabergoline CRS.
Test solution Dissolve 30.0 mg of the substance to be
STO RA G E
examined in the mobile phase and dilute to 25.0 m L with
Protected from light.
the mobile phase.
1-350 Caffeine 2016
IMPURITIES Content
Specified impurities A, B, C, D 98.5 per cent to 101.5 per cent (dried substance).
CHARACTERS
CH2
Appearance
W hite or almost white, crystalline powder or silky, white or
almost white, crystals.
Solubility
Sparingly soluble in water, freely soluble in boiling water,
slightly soluble in ethanol (96 per cent). It dissolves in
concentrated solutions of alkali benzoates o r salicylates.
A. ( 6 a/?,9/?, 10ai?)-7-(prop-2-enyl)-4,6,6a,7,8,9, 10,10a- It sublimes readily.
octahydroindolo [4,3 -fg] quinoline-9-carboxylic acid. IDENTIFICATION
First identification A , B, E
Second identification A , C, D, E, F.
A. M elting point (2.2.14): 234 °C to 239 °C.
B. Infrared absorption spectrophotometry (2.2.24).
r- n
Comparison caffeine CRS.
C. T o 2 m L of a saturated solution add 0.05 m L o f iodinated
potassium iodide solution R. T he solution remains clear.
Add 0.1 m L o f dilute hydrochloric add R; a brown precipitate
B. R = C O -N H -C 2H 5, R ' = H : (6ai? 39i?J10a/?)-iV9-[3-
is formed. Neutralise with dilute sodium hydroxide solution R\
(dimet±iylaxnino)propyI]-iV4-ethyl-7-(prop-2-enyl)-
the precipitate dissolves.
6a373839310jl Oa-hexahydroindolo [4 ,3 - ^ quinoline-4,9 (6H)-
dicarboxamide, D. In a ground-glass-stoppered tube, dissolve about 10 mg in
0.25 m L o f a mixture of 0.5 m L of acetylacetone R and 5 m L
C. R = R ' = C O -N H -C 2H 5: (6aÄ,9i? 310aÄ )-^ 9-[3-
o f dilute sodium hydroxide solution R. H eat in a water-bath at
(dimethylamino)propyl] -N 4-ethyl-iV 9-(et±ijdcaTbamoyl)-7 -
80 °C for 7 min. Cool and add 0.5 m L of
(prop-2-eriyl)-6a,7,83931031Oa-hexahydroindolo [4,3-
dimethylamiitobenzaldehyde solution R2. H eat again in a water-
fg] quinoline-4,9 (6/i)-dicarboxam ide,
bath at 80 °C for 7 min. Allow to cool and add 10 m L of
D. R = R ' = H: (6ai?, 9R, 1QaR)-N- [3- water R', an intense blue colour develops.
(dimethylamino)propyl] -7-(prop-2-enyl) -43636a,7,8,9,10,10a-
E. Loss on drying (see Tests).
octahydroindolo [4,3-fg] quinoline-9-carboxamide.
F. It gives the reaction o f xanthines (2.3.1).
PhEur
TESTS
Solution S
Dissolve 0.5 g with hearing in 50 m l. o f carbon dioxide-free
water R prepared from distilled water i?, cool and dilute to
★ ★
Caffeine ★ ★ 50 m L with the same solvent.
Anhydrous Caffeine ***** Appearance o f solution
Solution S is d ear (2.2.1) and colourless (2.2.2, Method II).
(Ph. Eur. monograph 0267)
A cid ity
T o 10 m L o f solution S add 0.05 m L of bromothymol blue
X solution R l; the solution is green or yellow. N o t more than
XX> 0.2 m L o f 0.01 M sodium hydroxide is required to change the
colour o f the indicator to blue.
I
ch 3 Related substances
Liquid chromatography (2.2.29).
C 8H 10N 4O 2 194.2 58-08-2 Test solution Dissolve 0.100 g of the substance to be
examined in the mobile phase and dilute to 50.0 m L with
Action and use the mobile phase. Dilute 1.0 m L o f this solution to 10.0 m L
Central nervous system stimulant. with the mobile phase.
Preparations Reference solution (a) Dilute 2.0 m L o f the test solution to
Aspirin and Caffeine Tablets 100.0 m L with the mobile phase. D ilute 1.0 m l. of this
Caffeine Citrate Injection solution to 10.0 mL with the mobile phase.
Caffeine C itrate Oral Solution Reference solution (b) Dissolve 5 m g o f caffdne for system
Paracetamol and Caffeine Capsules suitability CRS (containing impurities A, C, D and F ) in the
Paracetamol and Caffeine Tablets mobile phase and dilute to 5 m L with the mobile phase.
D ilute 2 m L o f this solution to 10 m L with the mobile
Paracetamol, Codeine Phosphate and Caffeine Capsules
phase.
Paracetamol, Codeine Phosphate and Caffeine Tablets Column:
PhEur______________________________________________________ — size: I = 0.15 m , 0 = 4.6 mm ;
— stationary phase: base-deactivated end-capped octadecylsilyl
DEFINITION silica gel for chromatography R (5 pm).
1,3,7 -Trimethyl-3,7 -dihydro-1H -purine-2, 6-dione.
2016 Caffeine 1-351
0
Mobile phase Mix 20 volumes of tetrahydrofuran R,
25 volumes of acetonitrile R and 955 volumes of a solution
containing 0.82 g/L of anhydrous sodium acetate R previously
adjusted to p H 4.5 with glacial acetic add R.
Flow rate 1.0 mL/min. ch 3
A c id ity
Caffeine Hydrate ***** T o 10 m l . o f solution S add 0.05 m L o f bromothymol blue
*. *
(Caffeine Monohydrate, Ph Eur monograph 0268) * solution R l; the solution is green or yellow. N o t m ore than
0.2 m L o f 0.01 M sodium hydroxide is required to change the
colour o f the indicator to blue.
Related substances
Liquid chromatography (2.2.29).
Test solution Dissolve 0.110 g o f the substance to be
I
ch 3 examined in the mobile phase and dilute to 50.0 m L with
the mobile phase. Dilute 1.0 m L of this solution to 10.0 m L
w ith the mobile phase.
CgHxoN.AjHiO 212.2 5743-12-4
Reference solution (a) Dilute 2.0 m L of the test solution to
A ctio n a n d u se 100.0 m L with the mobile phase. Dilute 1.0 m L of this
Central nervous system stimulant. solution to 10.0 m L with the mobile phase.
Reference solution (b) Dissolve 5 m g of caffeine for system
Ph Eur___________________________________________________________
suitability CRS (containing impurities A, C, D and F) in the
D E F IN IT IO N mobile phase and dilute to 5.0 m L with the mobile phase.
l,3,7-Trim ethyl-3,7-dihydro-lH -purine-2,6-dione D ilute 2.0 m L o f this solution to 10.0 m L with the mobile
monohydrate. phase.
C o n te n t Column:
98.5 per cent to 101.5 per cent (dried substance). — size. I = 0.15 m , 0 = 4.6 mm ;
— stationary phase, base-deactivated end-capped octadecylsHyl
CHARACTERS
silica gelfor chromatography R (5 Jim).
A p p e a ra n c e
Mobile phase Mix 20 volumes o f tetrahydrofuran R,
W hite or almost white, crystalline powder or silky, white or
25 volumes of acetonitrile R and 955 volumes o f a solution
alm ost white crystals.
c o n t a i n i n g 0.82 g/L of anhydrous sodium acetate R previously
S o lu b ility adjusted to p H 4.5 with glacial acetic add R.
Sparingly soluble in water, freely soluble in boiling water,
Flow rate 1.0 mlVmin.
slightly soluble in ethanol (96 per cent). It dissolves in
concentrated solutions o f alkali benzoates or salicylates. Detection Spectrophotom eter at 275 nm.
It sublimes readily. Injection 10 |iL.
Run time 1.5 times the retention time o f caffeine.
ID E N T IF IC A T IO N
First identification A, B, E. Identification of impurities U se the chrom atogram supplied
with caffeine for system suitability CRS and the chrom atogram
Second identification A, C, D, E, F. obtained with reference solution (b) to identify the peaks due
A. M elting point (2.2.14): 234 °C to 239 °C, determ ined to impurities A, C , D and F.
after drying at 100-105 °C.
Retention time Caffeine = about 8 min.
B. Infrared absorption spectrophotometry (2.2.24). System suitability: reference solution (b):
Preparation D ry the substance to be examined at 100-105 °C — resolution: minimum 2.5 between the peaks due to
before use. impurities C and D; m in im u m 2.5 between the peaks due
Comparison caffeine CRS. to impurities F and A.
C. T o 2 m L o f a saturated solution add 0.05 m L o f iodmazed Limits:
potassium iodide solution R; the solution remains clear. — unspecified impurities: for each impurity, no t m ore than
A dd 0.1 m L o f dilute hydrochloric add R; a brown precipitate 0.5 times the area of the principal peak in the
is formed. Neutralise with dilute sodium hydroxide solution R; chromatogram obtained with reference solution (a)
the precipitate dissolves. (0.10 per cent);
D . In a glass-stoppered tube, dissolve about 10 m g in — total: n o t more than 0.5 times the area of the principal
0.25 m L of a mixture of 0.5 m L of acetylacetone R and 5 m L peak in the chromatogram obtained with reference
o f dilute sodium hydroxide solution R. H eat in a w ater-bath at solution (a) ( 0.1 per cent);
80 °C for 7 min. Cool and add 0.5 m L of — disregard limit. 0.25 times the area o f the principal peak in
dxmethylaminobenzaldehyde solution R2. H eat again in a water- the chromatogram obtained with reference solution (a)
bath at 80 °C for 7 m in. Allow to cool and add 10 m L of (0.05 per cent).
water R; an intense blue colour develops. S u lfa te s (2.4.11)
E. Loss on drying (see Tests). M axim um 500 ppm , d e te rm in e d on 15 m L o f solution S.
F. It gives the reaction of xanthines (2.3.1). Prepare the standard using a mixture o f 7.5 m L o f sulfate
standard solution (10 ppm SO 4) R and 7.5 m l . of distilled
TESTS
water R.
S o lu tio n S
Dissolve 0.5 g with heating in 50 m L of carbon dioxide-free
Heavy m etals (2.4.8)
M axim um 20 ppm .
water R prepared from distilled water R, cool, and dilute to
50 m L with the same solvent. 1.0 g complies with test C. Prepare the reference solution
using 2 m L of lead standard solution (10 ppm Pb) R.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
2016 Calamine 1-353
L oss o n d ry in g (2.2.32)
5.0 per cent to 9.0 per cent, determined on 1.000 g by
h3c 1 -CHi
drying in an oven at 105 °C for 1 h. 'Hhn^X - >n
S u lfa te d a sh (2.4.14) I
ch3
Maximum 0.1 per cent, determined on 1.0 g.
A SSAY E. N ,l-dimethyl-4-(methylamino)-lff-imidazole-5-
Dissolve 0.170 g, previously dried at 100-105 °Cj with carboxamide (caffeidine).
heating in 5 m L o f anhydrous acetic acid R. Allow to cool, and
add 10 m L of acetic anhydride R and 20 m L of toluene R n Cl
T itrate with 0.1 M perchloric acid, determining the end-point
potentiometrically (2.2.20). Î Ï )
1 m L o f 0.1 M perchloric acid is equivalent to 19.42 mg H
o f C 8H 10N 4O 2.
IM P U R IT IE S F . l,7-dim ethyl-3,7-dihydro-lff-purine-2,6-dione.
Other detectable impurities (the following substances would, if PhEur
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use Calamine
(2034). It is therefore not necessary to identify these
Prepared Calamine
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): A , B, A ctio n a n d u se
C, D, E, F. Antipruritic.
P re p a ra tio n s
Aqueous Calamine Cream
h’ c ' n Calamine Lotion
O ^N Calamine Ointm ent
ch3 Calamine and Coal T a r Ointment
D E F IN IT IO N
A. l,3-dim ethyl-3,7-dihydro-lff-purine-2,6-dione
Calamine is a basic zinc carbonate suitably coloured with
(theophylline),
iron(m) oxide.
C H A R A C T E R IS T IC S
o y -H An amorphous, impalpable, pink o r reddish brown powder,
HjC'N-A y - NH the colour depending on the variety and amount of iron(m)
oxide present and the process by which it is incorporated.
0^ N NH! Practically insoluble in water. It dissolves with effervescence
ch 3 in hydrochloric acid.
ID E N T IF IC A T IO N
B. N -(6-am ino-l,3-dim ethyl-2,4-dioxo-l,2,3,4-
A. Yields the reactions characteristic of carbonates,
tetrahydropyrimidin-5-yl)formamide,
Appendix VI.
B. T o 2 g add 5 m L o f hydrochloric acid and heat to boiling;
if necessary, add hydrochloric acid drop wise until a bright
h 3c
yellow solution is obtained. Cool and add 13. 5m ammonia
or iV >
'n
1 I,
until the first sign of precipitate (solution A). T h e solution
yields reaction B characteristic o f iron salts, Appendix VI.
CHj Dilute 1 m L of solution A to 5 m L with water, the solution
yields the reaction characteristic o f zinc salts, Appendix VI.
C . 1,3,9-trimethyl-3,9-dihydro- lH -purine-2,6-dione TESTS
(isocaffeine),
C a lc iu m
Dissolve 0 .5 0 g in a mixture of 10 m L o f water and 2 .5 m l.
o f glacial acetic acid and filter. T o 0.5 m L of the filtrate add
1 'Cl 15 m L of 5 m ammonia and 2 m L of a 2 .5 % whr solution o f
x jC > ammonium oxalate and allow to stand for 2 minutes.
O' N T he solution remains clear.
1I
ch 3
S olu b le b a riu m salts
T o the remainder of the filtrate obtained in the test for
D . 3,7-dimethyl-3,7-dihydro-lff-purm e-2,6-dione Calcium add 2 m L o f 1m sulfuric acid and allow to stand for
(theobromine), 5 minutes. T he solution remains clear.
L ead
N o t more than 1 5 0 ppm when determined by atomic
absorption spectrophotometry, Appendix II D, M ethod II,
1-354 Calcifediol 2016
ASSAY
Liquid chromatography (2.2.29) as described in the test for
Anhydrous Calcipotriol *****
★ ★
related substances with the following modifications. (Ph. Eur. monograph 2011) *
Injection T est solution and reference solutions (a) and (c).
System suitability: reference solution (c):
— repeatability: maximum relative standard deviation of
1 per cent for the peak due to calcifediol after 6
injections.
Calculate the percentage content o f C 27H 44O 2 using the
chromatogram obtained with reference solution (a) and
taking into account the assigned content o f calcifedbl CRS
and, if necessary, the peak due to pre-caldfediol.
STO RA G E
U nder nitrogen, in an airtight container, protected from light,
at a tem perature o f 2 °C to 8 °C.
C jvK kA 412.6 112965-21-6
T he contents of an opened container are to be used
immediately. A ctio n a n d use
IM P U R IT IE S Vitamin D analogue.
Specified, impurities A, B, C, D. P re p a ra tio n s
Calcipotriol Cream
Calcipotriol Ointm ent
Calcipotriol Scalp Application
PhEur__________________________________________________________
D E F IN IT IO N
(5Z,7£,22£,245)-24-Cydopropyl-9,10-secochola-
5,7,10( 19),22-tetraene-l a,3P,24-triol.
A. 9 P, 10a-cholesta-5,7-diene-3 (3325-diol,
C o n te n t
95.5 per cent to 102.0 p er cent (dried substance).
A reversible isomérisation to pre-caldpotriol takes place in
solution, depending on tem perature and time. T h e activity is
due to both compounds.
CHARACTERS
A p p e a ra n c e
W hite or almost white, crystalline powder.
B. cholesta-5,7-diene-3P,25-diol, S o lubility
Practically insoluble in water, freely soluble in ethanol
(96 per cent), slightly soluble in methylene chloride.
It is sensitive to heat and light.
ID E N T IF IC A T IO N
A. Infrared absorption spectrophotometry (2.2.24).
Comparison Ph. Eur. reference spectrum of anhydrous calcipotriol.
B. Loss on drying (see Tests).
TESTS
Carry out the tests for related substances and the assay as rapidly
as possible and protected from actinic light and air.
C . (6£)-9,l0-secocholesta-5(l0),6,8-triene-3P,25-diol, R ela te d su b sta n ce s
A. Thin-layer chromatography (2.2.27).
Solution A T o 1 m L of triethylamine R add 9 m L o f
chloroform R.
Test solution Dissolve 1 m g of the substance to be examined
in 100 jiL of solution A.
Reference solution (a) T o 10 (iL o f the test solution add
990 jiL o f solution A.
Reference solution (b) T o 250 (iL o f reference solution (a) add
750 (iL o f solution A.
Reference solution (c) T o 100 (iL o f reference solution (a) add
900 pL of solution A.
D . (5£,7£)-9,l0-secocholesta-5,7,10( 19)-triene-3P,25-diol.
!____________:____________ Ph Eur
1-356 Anhydrous Calcipotriol 2016
Reference solution (d) Place 2 mg o f the substance to be Run time Twice the retention time of caldpotriol.
examined in a vial and dissolve in 200 (iL of solution A. Relative retention W ith reference to caldpotriol (retention
Close the vial and keep it in a water b ath at 60 °C for 2 h. time = about 13.5 min): im purity B = about 0.86;
Plate TLC silica gel F2 $ 4 plate R. impurity C = about 0.92; impurity D = about 1.3.
Mobile phase 2-methylpropanol R, methylene chloride R System suitability: reference solution (c):
(20:80 VIV). — peak-UHvaHey ratio: m inim um 1.5, where Hp = height
Application 10 [iL of the test solution and reference above the baseline o f the peak due to im purity C and
solutions (b), (c) and (d). Hv = h d g h t above the baseline of the lowest point of
the curve separating this peak from the peak due to
Development Over 2/3 of the plate.
caldpotriol;
Drying In air, then at 140 °C for 10 min. — the chrom atogram obtained is similar to the
Detection Spray the h o t plate with an alcoholic solution of chrom atogram supplied with caldpotriol
sulfuric add R, dry at 140 °C for not m ore than 1 m in and monohydrate CRS.
examine in ultraviolet light a t 366 nm . Limits:
Relative retention With reference to calcipotriol — impurity B: not m ore than 0.5 times the area of the
(Rp = about 0.4): impurity G = about 0.4; principal peak in the chrom atogram obtained with
im purity H = about 0.4; pre-caldpotriol = about 0.9; reference solution (a) (0.5 per cent);
impurity A = about 1.2 . — impurities C, D: for each impurity, not m ore than the
System suitability Reference solution (d): area of the prindpal peak in the chromatogram
— the chrom atogram shows a secondary spot due to pre- obtained with reference solution (a) ( 1.0 per cent);
caldpotriol. — unspecified impurities: for each impurity, n o t m ore than
Limits: the area o f the principal peak in the chrom atogram
— impurity A: any spot due to im purity A is not more obtained with reference solution (b) (0.10 per cent);
intense than the spot in the chrom atogram obtained — total: not m ore than 2.5 times the area o f the prindpal
with reference solution (b) (0.25 per cent); peak in the chrom atogram obtained with reference
— impurities G, H: any spot due to im purity G or H is solution (a) (2.5 p er cent);
not m ore intense than the spot in the chromatogram — disregard lim it 0.5 times the area o f the prindpal peak
obtained with reference solution (b) (0.25 per cent for in the chrom atogram obtained with reference
the sum ); solution (b) (0.05 per cent).
— unspecified impurities: any other spot is not m ore Loss on drying
intense than the spot in the chrom atogram obtained M aximum 1.0 p er cent, determ ined on 5 m g by
with reference solution (c) (0.1 per cent). thermogravimetry (2.2.34). H eat to 105 °C at a rate of
B. Liquid chrom atography (2.2.29). 10 °C/min and maintain at 105 °C for 60 min.
Solution A Dissolve 1.32 g o f ammonium phosphate R in ASSAY
water R and dilute to 10.0 m L with th e same solvent. Liquid chromatography (2.2.29) as described in the test for
Solvent mixture Solution A, water R, methanol R related substances with the following modification.
(0.3:29.7:70 V/V/V). Injection T est solution (b) and reference solution (d).
Test solution (a) Dissolve 2 m g of the substance to be Calculate the percentage content o f C 27H 40O 3 taking into
examined in the solvent mixture and dilute to 5.0 m L with account the assigned content o f caldpotriol monohydrate CRS.
the solvent mixture.
STO RA G E
Test solution (b) Dissolve 2.00 mg o f the substance to be In an airtight container, protected from light, at —20 °C or
examined in the solvent mixture and dilute to 20.0 m L with
below.
the same solvent mixture.
IM P U R IT IE S
Reference solution (a) D ilute 1.0 m L o f test solution (a) to
100.0 m L with the solvent mixture. Spedfied impurities A, B, C , D , G , H
Reference solution (b) Dilute 1.0 m L o f reference solution (a) Other detectable impurities (the following substances would, if
to 10.0 m L w ith the solvent mixture. present at a suffident level, be detected by one or other o f
the tests in the m onograph. T hey are limited by the general
Reference solution (c) Dissolve 1 mg o f caldpotriol
acceptance criterion for other/unspecified impurities and/or
monohydrate CRS (containing impurities B, C and D ) in the
by the general monograph Substances for pharmaceutical use
solvent m ixture and dilute to 2.5 m L w ith the solvent
(2034). It is therefore not necessary to identify these
mixture.
impurities for dem onstration o f compliance. See also 5.10.
Reference solution (d) Dissolve 2.00 m g o f caldpotriol Control of impurities in substances for pharmaceutical use): E, F,
monohydrate CRS in the solvent mixture and dilute to I.
20.0 m L with the solvent mixture.
By thin-layer chromatography: A , G, H, I.
Column:
— size: I = 0.10 m , 0 = 4.0 mm ;
By liquid chromatography: B, C, D, E, F.
— stationary phase: octadecylsUyl silica gel for
chromatography R (3 Jim).
Mobile phase water R, methanol R (30:70 V!V).
Flow rate 1.0 mL/min.
Detection Spectrophotom eter at 264 nm .
Injection 20 |iL of test solution (a) and reference solutions (a),
(b) and (c).
2016 Anhydrous Calcipotriol 1-357
C. (5£j7£j22£j245)-24-cyclopropyl-9j 10-secochola-
5,7,10(19)j22-tetraene-laj3ßj24-ttiol ((5£)-caldpotriol),
D . (5Z,7Ẹ,22.Ẹ,24j?!)-24-cyclopropyl-9,10-secochola-
5,7,10( 19),22-tetraene-1aj3ß,24-triol (24-ẹpt-calcipotriol),
H . (5Zj7£j22jEj24R)-24-cyclopropyl-24-[[(5Zj7£j22jEj245)-
24-cydopropyl-la,3ß-dihydroxy-9,10-secochola-
5 ,7 ,10(19),22-tetraen-24-yl]oxy]-9,10-secochola-
5,7,10(19) j22-tetraène-l aj3ß-diolj
1-358 Calcipotriol Monohydrate 2016
TESTS
Carry out the tests for related substances and the assay as rapidly
as possible and protected from actinic light and air.
Related substances
A. Thin-layer chromatography (2.2.27).
Solution A T o 1 m L of triethylamine R add 9 m l. o f
chloroform R.
Test solution Dissolve 1 mg of the substance to be exam in ed
in 100 pL o f solution A.
Reference solution (a) T o 10 |iL o f the test solution add
I. (65’j7iîj8i?j22£j245)-24-cyclopropyl-6j8:7j 19-dicyclo-9,10-
990 |jL o f solution A.
secochola-5(10),22-diene-la,3P,24-triol (suprasterol of
calcipotriol). Reference solution (b) T o 250 |iL of reference solution (a) add
750 pL o f solution A.
___________________________________________________________PhEur
Reference solution (c) T o 100 |iL o f reference solution (a) add
900 jjL o f solution A.
Reference solution (d) Place 2 mg o f the substance to be
examined in a vial and dissolve in 200 |iL o f solution A.
Calcipotriol Monohydrate Close the vial and keep it in a water bath at 60 °C for 2 h.
(Ph. Eur. monograph 2284) * Plate TLC silica gel F2 5 4 plate R
Mobile phase 2-methylpropanol R, methylene chloride R
(20:80 VIV).
Application 10 (iL o f the test solution and reference
solutions (b), (c) and (d).
Development Over 2/3 of the plate.
Drying In air, then at 140 °C for 10 min.
Detection Spray the hot plate with an alcoholic solution of
sulfuric acid R, dry at 140 °C for n o t more than 1 min and
examine in ultraviolet light at 366 nm.
Relative retention W ith reference to calcipotriol
(Rp = about 0.4): impurity G = about 0.4;
im purity H = about 0.4; pre-calcipotriol = about 0.9;
C 27H 40O 332 O 430.6 147657-22-5 im purity A = about 1.2.
Reference solution (c) Dissolve 1 mg of calcipotriol (2034). It is therefore not necessary to identify these
monohydrate CRS (containing impurities B, C and D ) in the impurities for demonstration of compliance. See also 5.10.
solvent mixture and dilute to 2.5 m L with the solvent Control of impurities in substances for pharmaceutical use): E, F,
mixture. I.
Reference solution (d) Dissolve 2.00 m g of calcipotriol By thin-layer chromatography: A, G, H, I.
monohydrate CRS in the solvent mixture and dilute to By liquid chromatography: B, C, D, E, F.
20.0 m L with the solvent mixture.
Column:
— size: I = 0.10 m, 0 = 4.0 mm;
— stationary phase: octadecylsOyl silica gel for
chromatography R (3 |im).
Mobile phase water R, methanol R (30:70 V!V).
Flow rate 1.0 mL/min.
Detection Spectrophotometer at 264 nm .
Irgection 20 p L o f test solution (a) and reference solutions (a),
(b) and (c).
Run time Twice the retention time o f calcipotriol.
Relative retention W ith reference to calcipotriol (retention A. (5Z,7£,22JB)-24-cyclopropyl-1a,3P-dihydroxy-9,10-
time = about 13.5 min): impurity B = about 0.86; secochola-5,7,10(19),22-tetraen-24-one,
impurity C = about 0.92; impurity D = about 1.3.
System suitability: reference solution (c):
— peak-to-valley ratio: minimum 1.5, where Hp = height
above the baseline o f the peak due to impurity C and
Hv = height above the baseline o f the lowest point of
the curve separating this peak from the peak due to
caldpotriol;
— the chromatogram obtained is similar to the
chromatogram supplied with calcipotriol
monohydrate CRS.
Limits:
— impurity B: not more than 0.5 times the area of the
principal peak in the chromatogram obtained with
reference solution (a) (0.5 per cent); B. (5Z,7Z,22£,245)-24-cydopropyl-9,10-secochola-
5,7,10(19),22-tetraene-1a,3p,24-triol ((7Z)-calcipotriol),
— impurities C, D: for each impurity, n o t more than the
area of the principal peak in the chromatogram
obtained with reference solution (a) ( 1.0 per cent);
— unspecified impurities: for each impurity, not more than
the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.10 per cent);
— total: n o t more than 2.5 times the area o f the principal
peak in the chromatogram obtained with reference
solution (a) (2.5 per cent);
— disregard limic. 0.5 times the area of the principal peak
in the chromatogram obtained with reference
solution (b) (0.05 per cent).
Water (2.5.12)
3.3 per cent to 5.0 per cent, determined on 0.100 g . C. (5£,7£,22£,245)-24-cydopropyl-9,10-secochola-
5,7,10(19),22-tetraene-la,3p,24-triol ((5£)-calcipotriol),
A SSAY
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modification.
Injection T est solution (b) and reference solution (d).
Calculate the percentage content of C 27H 40O 3 taking into
account the assigned content of calcipotriol monohydrate CRS.
STO RA G E
In an airtight container, protected from light.
IM P U R IT IE S
Specified impurities A, B, C, D , G, H
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of D. (5Z,7£,22£,24i?)-24-cyclopropyl-9,10-secochola-
the tests in the monograph. They are limited by the general 5,7,10(19),22-tetraene-1otJ3p,24-triol (24-ept-calcipotriol),
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
1-360 Calcitonin (Salmon) 2016
H H
I. (65’37i?,8i?,22£'j245)-24-cydopropyl-6,8:7,19-dicyclo-9,10-
E. rac-(5Z,7J5'j245)-24-cyclopropyl-9,10-secochola-
secochola-5 (10),22-diene-10,3(3,24-triol (suprasterol o f
5,7,10(19)-triene-1a,3 P,24-triol,
calapotriol).
___________________________________________________________PhEur
H- Cys - Ser - Asn - Leu- Ser - Thr - Cys - Val - Leu - Gly-
l_______________________ i
h3c ch3 h3c ch3 Lys - Leu - Ser - Gin- Glu - Leu—His - Lys - Leu - Gin-
Thr - Tyr - Pro - Arg - Thr - Asn - Thr - Gly - Ser - Gly-
F. (5Z, 7£,222j,24S)-24-cyclopropyl-1a ,3 P-bis [[(1,1-
Thr-Pro-NH2
dimethylethyl)dimethylsilyl] oxy]-9,10-secochola-
5,7,10(19),22-tetraen-24-ol,
C 145H 240N 44O 48S2 3432 47931-85-1
PhEur_______________________________________________________________
DEFINITION
Polypeptide having the structure determ ined for salm on
caldtonin I. It lowers the caldum concentration in plasma of
mammals by dim in ishin g the rate o f bone resorption. It is
obtained by chemical synthesis or by a m ethod based on
recom binant D N A (rDNA) technology. It is available as an
acetate.
Content
G . 24,24 '-oxybis [(5Z,7£32 2 £ 3245)-24-cydopropyl-9j 10- 90.0 per cent to 105.0 per cent o f the peptide
secochola-5,7,10( 19) ,22-tetxaene-1a,3 P-diol], C 145H 240N 44O 48S 2 (anhydrous and acetic add-free
substance).
By convention, for the purpose o f labelling caldtonin
(salmon) preparations, 1 mg o f caldtonin (salmon)
(C j 45H 240N 44O 48S 2) is equivalent to 6000 IU o f biological
activity.
PRODUCTION
T h e following requirements apply only to caldtonin (salmon)
produced by a m ethod based on rD N A technology.
Prior to rdease the following tests are carried out on each
batch o f final bulk product unless exemption has been
granted by the competent authority.
Host-cell-derived proteins
T h e limit is approved by the competent authority.
H ost-cell or vector-derived DNA
T h e limit is approved by the com petent authority.
H . (5Z,72j,222j,24i?)-24-cyclopropyl-24- [ [(5Zj7£’j22£,245) -
24-cyclopropyl-1a,3 (J-dihydroxy-9,1O-secochola- CHARACTERS
5,7,10(19),22-tetraen-24-yl] oxy]-9,10-secochola- Appearance
5,7,10(19),22-tetraene-1a,3|3-diol, W hite or alm ost white powder.
2016 Calcitonin (Salmon) 1-361
15.1 - 22.1 100 0 H - Cys - Ser - Asn - Leu - Ser - Thr - Cys - Val - Leu - Gly-
1------------------------------------------------------- 1 10
Lys - Leu - Ser - Gin - Glu - Leu - His - Lys - Leu - Gin-
20
Flow rate 1.2 mL/min. Thr - Pro - Arg - T h r-A s n -T h r-G ly -S e r -G ly -T h r -
30
Detection Spectrophotom eter at 220 nm . Pro-NH2
Injection 50 pL; rinse the injector with a 40 p er cent V/V
solution o f acetomtrUe for chromatography R. C. des-22-tyrosine-caldtonin (salmon),
Relative retention W ith reference to calcitonin (salmon) D . O-acetyiated caldtonin (salmon),
(retention tim e = about 9 min): impurity G = about 0.4;
impurity F = about 0.6; impurity E = about 0.9.
System suitabUity Resolution solution:
— resolution: minimum 3.0 between the peaks due to
im purity E and calcitonin (salmon).
2016 Calcitriol 1-363
s o 3h s o 3h
3I 3I TESTS
H- Ala - Ser - Asn - Leu - Ser - Thr - Ala - Val - Leu - Gly- Related substances
10
Lys - Leu - Ser-Gln-Glu - Leu - His - Lys - Leu - Gin - Liquid chromatography (2.2.29): use the normalisation
20
Thr - Tyr - Pro - Arg - Thr - Asn - Thr - Gly - Ser - Gly- procedure. Carry out the test as rapidly as possible, avoiding
30 exposure to actinic light and air.
Thr - Pro-R
Test solution Dissolve 1.00 mg o f the substance to be
exam in ed without heating in 10.0 m L of the mobile phase.
F. R = N H 2: [lj7-bis(3-sxalfo-L-alanme)]caldtonin (salmon),
Reference solution (a) Dissolve 1.00 m g o f caldtriol CRS
G . R = N H -C H 2- C 0 2H: [l,7-bis(3-sulfo-L-
w ithout heating in 10.0 m L of the mobile phase.
alanine)]caldtoninylglydne (salmon).
Reference solution (b) Dilute 1.0 m l. o f reference solution (a)
___________________________________________________________________________________________ Ph Eur
to 100.0 m L with the mobile phase. Dilute 1.0 m L of this
solution to 10.0 m L with the mobile phase.
Reference solution (c) H eat 2 m L o f reference solution (a) at
80 °C for 30 min.
★ ★
Calcitriol ★ ★ Column:
**+★* — size: I = 0.25 m, 0 = 4.6 mm;
(Ph. Eur. monograph 0883) — stationary phase: octylsUyl silica gel for chromatography R1
(5 pm);
H3C H — temperature: 40 °C.
Mobile phase Mix 450 volumes o f a 1.0 g/L solution of
H V -C H 3
tris(kydroxymetkyl)aminomethane R adjusted to p H 7.0-7.5
IX y HO ch3 with phosphoric add R, and 550 volumes o f acetonitrile R.
I *■* Flow raxe 1.0 mL/min.
Detection Spectrophotometer at 230 nm.
J L ^ ch2 Injection 50 |xL.
H O -y k ^ J^ O H Run time Twice the retention time o f caldtriol.
H H Relative retention W ith reference to caldtriol (retention
time = about 14 min): impurity C = about 0.4; pre-
416.6 32222-06-3 caldtriol = about 0.88; impurity A = about 0.95;
impurity B = about 1.1.
Action and use System suitability:
Vitamin D analogue. — resolution: m inim um 3.5 between the peaks due to pre-
Preparation caldtriol and caldtriol in the chromatogram obtained with
C aldtriol Capsules reference solution (c);
— number of theoretical plates: m inim u m 10 000, calculated
P h E ir ____________________________________________________________________ for the peak due to caldtriol in the chromatogram
D E F IN IT IO N obtained with reference solution (a).
(5Z,7£)-9,10-Secocholesta-5,7,10(19)-triene-l a,3P,25-triol. Limits:
Content — impurities A , B, C: for each impurity, maximum
0.5 per cent;
97.0 per cent to 103.0 per cent.
— unspecified impurities: for each impurity, maxim um
A reversible isomerisation to pre-caldtriol takes place in
0.10 per cent;
solutipn, depending on temperature and time. T he activity is
— total: maximum 1.0 per cent;
due to b oth com pounds (see Assay).
— disregard limit:. 0.5 times the area o f the prindpal peak in
CHARACTERS the chromatogram obtained w ith reference solution (b)
A p p e a ra n c e (0.05 per cent); disregard the peak due to pre-caldtriol.
White or almost w hite crystals. ASSAY
Solubility Liquid chromatography (2.2.29) as described in the test for
Practically insoluble in water, freely soluble in ethanol rd ated substances with the following modifications.
(96 per cent), soluble in fatty oils. Injection T est solution and reference solution (a).
It is sensitive to air, heat and light. System suitability: reference solution (a):
ID E N T IF IC A T IO N — repeatability: maximum relative standard deviation of
A. Infrared absorption spectrophotometry (2.2.24). 1 per cent for die peak due to caldtriol after 6 injections.
Comparison Ph. Eur. reference spectrum of caldtriol. Calculate the percentage content o f Q 7H 44O 3 taking into
account the assigned content o f calcitriol CRS and, if
B. Examine the chromatograms obtained in the assay.
necessary, the peak due to pre-caldtriol.
Results T h e prindpal peak in the chromatogram obtained
with the test solution is similar in retention time and size to STO RA G E
the prindpal peak in the chromatogram obtained with U nder nitrogen, in an airtight container, protected from light,
reference solution (a). at a tem perature of 2 °C to 8 °C.
T he contents o f an opened container are to be used
immediately.
1-364 Calcium Acetate 2016
IM P U R IT IE S
Specified impttrities A, B, C
Calcium Acetate *****
** +*
(Calcium Acetate, Anhydrous, *
Ph Ever monograph 2128)
Ca2+ [h 3C - C 0 2-]2
C ^C aO * 158.2 62-54-4
A c tio n a n d u se
U sed in solutions for haemodialysis and peritoneal dialysis.
PhEur__________________________________________________________
D E F IN IT IO N
A. (5E,7E)-9,l0-secocholesta-5j7jl0( 19)-triene-laj3ß,25-triol
(rrans-calcitriol). C ald u m diacetate.
C o n te n t
99.0 per cent to 101.0 per cent (anhydrous substance).
CHARACTERS
A p p e a ra n c e
W hite or almost white, hygroscopic powder.
S o lu b ility
Freely soluble in water, slightly soluble in ethanol
(96 per cent).
ID E N T IF IC A T IO N
A. It gives reaction (b) of caldum (2.3.1).
B. It gives reaction (b) of acetates (2.3.1).
B. (5Zj7£)-9j 10-secocholesta-5j7j 10(19)-triene-l ß,3ß,25-triol TESTS
(lß-caldtriol), S o lu tio n S
Dissolve 5.0 g in carbon dioxide-free water R and dilute to
50.0 m L w ith the same solvent.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
p H (2.2.3)
7.2 to 8.2.
Dilute 5.0 m L o f solution S to 10.0 m L with carbon dioxide-
free water R.
R e ad ily o x id isab le su b stan ces
Dissolve 2.0 g in boiling water R and dilute to 100 m L with
boiling water R, add a few glass beads, 6 m L of 5 M sulfuric
C. (6aR,72?,9a/?)-l l-[(3S,5.R)-3,5-dihydroxy-2- acid and 0.3 m L of 0.02 Mpotassium permanganate, mix, boil
m ethylcyclohex-l-enyl]-7-[(lJ?)-5-hydroxy-l,5- gently for 5 min and allow the predpitate to settle. T h e pink
dim ethylhexyi]-6a-methyl-2-phenyl-5,6,6a,7,8,9,9a,l 1- colour in the supernatant is not completely discharged.
octahydro-l//j4aii-cy d o p en ta \f\ [1,2,4] triazolo [1,2- C h lo rid e s (2.4.4)
a]cinnoline-1,3 (2/i)-dione (triazoline adduct o f pre- M aximum 330 ppm.
caldtriol).
Dissolve 0.15 g in water R and dilute to 15 m L with the
___________________________________________________________PhEur same solvent.
F lu o rid e s
M aximum 50 ppm.
Potentiom etry (2.2.36, Method I).
Test solution In a 50 m L volumetric flask, dissolve 0.200 g in
a 10.3 g/L solution of hydrochloric acid R, add 5.0 m L of
fluoride standard solution (1 ppm F) R and dilute to 50.0 m L
with a 10.3 g/L solution o f hydrochloric add R. T o 20.0 m L
of the solution add 20.0 m L of total-ionic-strength-adjiistment
buffer R and 3 m L o f an 82 g/L solution of anhydrous sodium
acetate R. Adjust to pH 5.2 with ammonia R and dilute to
50.0 m L with distilled water R.
Reference solutions T o 0.25 m l^ 0.5 mL, 0.75 m L and
1.0 m l. of fluoride standard solution (10 ppm F) R add
20.0 m L o f total-ionic-strength-adjustment buffer R and dilute to
50.0 m L w ith distilled water R.
2016 Calcium Acetate 1-365
Indicator electrode Fluoride selective. Reference solutions Prepare the reference solutions using
Reference electrode Silver-silver chloride. potassium standard solution (0.2 per cent K) R, diluted as
necessary with water R.
Take into account the addition of fluoride to the test solution
for the calculation. Wavelength 766.5 nm.
N itra te s S o d iu m
T o 10.0 m L o f solution S add 5 m g o f sodium chloride R, M aximum 500 ppm , if intended for use in the m anufacture
0.05 m L o f indigo carmine solution R and add with stirring, o f parenteral preparations or haemodialysis solutions.
10 m L o f nitrogen-free sulfuric acid R. T h e blue colour Atomic emission spectrometry (2.2.22, Method II).
remains for at least 10 min. Test solution Dissolve 1.00 g of the substance to be examined
S u lfates ( 2.4.13) in water R and dilute to 100.0 m L with the same solvent.
Maximum 600 ppm. Reference solutions Prepare the reference solutions using sodium
Dissolve 0.25 g in distilled water R and dilute to 15 m L with standard solution (200 ppm Na) R, diluted as necessary with
the same solvent. water R.
A lu m in iu m (2.4.17) Wavelength 589 nm.
Maximum 1 ppm , if intended for use in the manufacture of S tro n tiu m
peritoneal dialysis solutions, haemofiltration solutions or M aximum 500 ppm , if intended for use in the m anufacture
haemodialysis solutions. o f parenteral preparations or haemodialysis solutions.
Test solution Dissolve 4.0 g o f the substance to be examined Atomic emission spectrometry (2.2.22, Method II).
in 100 m l, o f water R and add 10 m L of acetate buffer solution Test solution Dissolve 2.00 g of the substance to be examined
pH 6.0 R. in water R and dilute to 100.0 m L with the same solvent.
Reference solution Mix 2 m L of aluminium standard solution Reference solutions Prepare the reference solutions using
(2 ppm AQ R, 10 m L of acetate buffer solution pH 6.0 R and strontium standard solution (1.0 per cent Sr) R, diluted as
98 m L o f water R. necessary with water R.
Blank solution M ix 10 m L o f acetate buffer solution pH 6.0 R Wavelength 460.7 nm.
and 100 m L o f water R.
H eav y m e ta ls (2.4.8)
A rsen ic (2.4.2) M aximum 10 ppm.
M axim um 3 ppm.
Dissolve 4.0 g in water R and dilute to 20 m L w ith the same
3.3 m L o f solution S complies with test A. solvent. 12 m L o f the solution complies with test A. Prepare
B a riu m the reference solution using lead standard solution
M aximum 50 ppm . (2 ppm Pb) R.
Inductively coupled plasma-atomic emission spectrometry W a te r (2.5.12)
(2.2.57). M axim um 7.0 per cent, determined on 0.100 g. A dd 2 m L
Test solution Dissolve 5.00 g of the substance to be examined of anhydrous acetic add R to the titration vessel in addition to
in water R and dilute to 100.0 m L w ith the same solvent. the methanol. Clean the titration vessel after each
Reference solutions Prepare the reference solutions using determination.
barium standard solution (0.1 per cent Ba) R, diluted as ASSAY
necessary with water R Dissolve 0.150 g in 100 m L of water R and carry out the
Wavelength 455.4 run. complexometric titration of calcium (2.5.11).
Ir o n (2.4.9) 1 m L o f 0.1 M sodium edetate is equivalent to 15.82 mg
Maximum 20 ppm , if intended for use in the manufacture o f o f C 4HôCa 0 4.
parenteral preparations or haemodialysis solutions. STORA GE
Dilute 5 m L o f solution S to 10 m L o f water R. In an airtight container.
M a g n e s iu m L A B E L L IN G
M axim um 500 ppm. T h e label states, where applicable, that the substance is
Atomic absorption spectrometry (2.2.23, Method II). suitable for u s e in the manufacture o f parenteral
Test solution Dissolve 50.0 m g of the substance to be preparations, peritoneal dialysis solutions, haemoflltration
exam in ed in water R and dilute to 100.0 m L with the same solutions or haemodialysis solutions.
solvent. __________________________________________________________ PhEur
Reference solutions Prepare the reference solutions using
magnesium standard solution (0.1 per cent Mg) R, diluted as
necessary with water R.
Source M agnesium hollow-cathode lamp.
Wavelength 285.2 nm.
Atomisation device Air-acetylene flame.
P o ta s s iu m
Maximum 500 ppm , if intended for use in the manufacture
o f parenteral preparations or haemodialysis solutions.
Atomic emission spectrometry (2.2.22, Method II).
Test solution Dissolve 1.00 g o f the substance to be examined
in water R and dilute to 25.0 mL w ith the same solvent.
1-366 Calcium Ascorbate 2016
Reference solution Mix 2 m L o f aluminium standard solution C. It complies with the limits of the assay.
(2 ppm Al) R, 10 m L o f acetate buffer solution pH 6.0 R and TESTS
98 m L of water R. S o lu tio n S
Blank solution M ix 10 m L of acetate buffer solution pH 6.0 R Dissolve 15.0 g in carbon dioxide-free water R prepared from
and 100 m L o f water R. distilled water R and dilute to 100 m L with the same solvent.
B a riu m A p p e a ra n c e o f solution
T o 10 m L o f solution S add 1 m L of calcium sulfate Solution S is clear (2.2.1) and n o t more intensely coloured
solution R. After at least 15 min, any opalescence in the th an reference solution Y6 (2.2.2, Method II).
solution is n o t more intense than that in a mixture of 1 m L Acidity or alkalinity
of distilled water R and 10 m l, o f solution S. T o 10 m L o f freshly prepared solution S add 0.1 m L of
Ir o n (2.4.9) phenolphthalein solution R. If the solution is red, not more
M aximum 10 ppm , determined on solution S. than 0.2 m L of 0.01 M hydrochloric add is required to
M a g n e s iu m a n d a lk ali m e ta ls discharge the colour and if the solution is colourless, n o t
M aximum 0.5 per cent. m ore than 0.2 m L o f 0.01 M sodium hydroxide is required to
tu rn it red.
T o a mixture o f 20 m L of solution S and 80 m L of water R
add 2 g of ammonium chloride R and 2 m l. of dilute S u lfa te s (2.4. IS)
ammonia R l, heat to boiling and pour into the boiling Maximum 200 ppm.
solution a h o t solution o f 5 g of ammonium oxalate R in D ilute 5 m L of solution S to 15 m L w ith distilled water R.
75 m L of water R. Allow to stand for 4 h, dilute to 200 m L A lu m in iu m
with water R and filter through a suitable filter. T o 100 m L T o 10 m L o f solution S add 2 m L of ammonium chloride
of the filtrate add 0.5 m L of sulfuric add R. Evaporate to solution R and 1 m l. of dilute ammonia R l. H eat to boiling.
dryness on a water-bath and ignite to constant mass at N o turbidity or precipitate is formed.
600 ± 50 °C. T he residue weighs a maximum of 5 mg
I f intended for use in the m anufacture o f dialysis solutions,
H eav y m e ta ls (2.4.8) the above test is replaced by the following test for aluminium
Maximum 20 ppm. (2.4.17): maximum 1 ppm.
12 m L of solution S complies with test A. Prepare the Prescribed solution Dissolve 6 g in 100 m L o f water R and add
reference solution using lead standard solution (2 ppm Pb) R. 10 m L o f acetate buffer solution pH 6.0 R.
A SSA Y Reference solution Mix 2 m L o f aluminium standard solution
Dissolve 0.280 g in 100 m L o f water R and carry out the (2 ppm Al) R, 10 m L of acetate biffer solution pH 6.0 R and
complexometric titration of calcium (2.5.11). 98 m L of water R.
2016 Calcium Dobesilate 1-369
Blank solution M ix 10 m L of acetate buffer solution pH 6.0 R Test solution Dissolve 0.100 g in water R and dilute to
and 100 m L of water R 200.0 m L with the same solvent. D ilute 5.0 m L o f this
B a riu m solution to 100.0 m L with water R.
T o 10 m L o f solution S add 1 m L o f calcium sulfate Spectral range 210-350 nm.
solution R. After at least 15 min, any opalescence in the Absorption maxima At 221 nm and 301 nm.
solution is not m ore intense than th at in a mixture of 1 m L Specific absorbance at the absorption maximum at 301 nm
of distilled water R and 10 m L of solution S. 174 to 181.
I r o n (2.4.9) B. M ix 1 m L o f ferric chloride solution R2, 1 m L o f a freshly
M axim um 7 ppm , determined on solution S. prepared 10 g/L solution of potassium ferricyanide R and
M a g n e s iu m a n d alk ali m etals 0.1 m L o f nitric add R. T o this mixture add 5 m L of freshly
M axim um 0.3 p e r cent. prepared solution S (see Tests): a blue colour and a
T o a mixture of 20 m L of solution S and 80 m L o f water R precipitate are immediately produced.
add 2 g of ammonium chloride R and 2 m L o f dilute ■C. 2 m L of freshly prepared solution S gives reaction (b) o f
ammonia R l, h eat to boiling and pour into the boiling calcium (2.3.1).
solution a h o t solution of 5 g of ammonium oxalate R in TESTS
75 m L of water R. Allow to stand for 4 h, dilute to 200 m L S o lu tio n S
with water R and filter through a suitable filter. T o 100 m L
Dissolve 10.0 g in carbon dioxide-free water R and dilute to
o f the filtrate add 0.5 m L of sulfuric add R. Evaporate to 100 m L with the same solvent.
dryness on a w ater-bath and ignite to constant mass at
600 ± 50 °C. T h e residue weighs a maximum o f 5 mg. A p p e a ra n c e o f so lu tio n
Solution S, when freshly prepared, is clear (2.2.1) and
H eav y m e ta ls (2.4.8) colourless (2.2.2, Method II).
M axim um 15 ppm .
p H (2.2.3)
12 m L o f solution S complies with test A. Prepare the
4.5 to 6.0 for solution S.
reference solution using lead standard solution (2 ppm Pb) R.
R e la te d su b sta n c e s
A SSAY Liquid chromatography (2.2.29). Keep all solutions at 2-8 °C.
Dissolve 0.200 g in 100 m L o f water R. Carry out the
Buffer solution Dissolve 1.2 g o f anhydrous sodium dihydarogen
complexometric titration of calcium (2.5.11).
phosphate R in 900 m L o f waterfor chromatography R, adjust
1 m L of 0.1 M sodium edetate is equivalent to 21.91 mg to p H 6.5 with disodium hydrogen phosphate solution R and
o f CaCl2,6H20 . dilute to 1000 m L with waterfor chromatography R.
L A B E L L IN G Test solution Dissolve 0.100 g o f die substance to be
T he label states, where applicable, that the substance is exam in e d in water R and dilute to 10.0 m L with the same
suitable for use in the manufacture o f dialysis solutions. solvent.
Ph Eur Reference solution (a) Dilute 1.0 m L of the test solution to
100.0 m L with water R Dilute 1.0 m L of this solution to
10.0 m L with water R.
Reference solution (b) Dissolve 10 m g of the substance to b e
***** ex a m in ed and 10 mg of hydroquinone R (impurity A) in
Calcium Dobesilate Monohydrate ★ ★ water R and dilute to 10 m L with the same solvent. Dilute
(Ph Eur monograph 1183) *★ * 1 m l . o f this solution to 100 m l . with water R.
Column:
— size: 1 = 0.25 m , 0 = 4.6 mm ;
Caz
XX OH
, H20
— stationary phase: spherical end-capped octadecylsûyl silica gel
for chromatography R (5 (im).
M obile phase acetonitrile R l , buffer solution (10:90 V IV ).
Flow rate 0.8 mL/min.
C 12H 10CaO10S*H 2O 436.4 20123-80-2 Detection Spectrophotometer at 220 nm.
PhEir__________________
Irtjection 10 |jL.
D E F IN IT IO N Run time 2.5 times the retention time o f dobesilate.
Calcium di(2,5-dihydroxybenzenesulfonate) monohydrate. Relative retention W ith reference to dobesilate (retention
C o n te n t time = about 6 min): impurity A = about 1.7.
99.0 per cent to 101.0 per cent (anhydrous substance). System suitability: reference solution (b):
CHARACTERS — resolution: m inim um 8.0 between the peaks due to
dobesilate and impurity A.
A p p e a ra n c e
W hite or almost white, hygroscopic powder. Limits:
— correction factor, for the calculation o f content, multiply the
S o lu b ility
peak area o f im purity A by 0.6;
Very soluble in water, freely soluble in anhydrous ethanol,
— impurity A: n o t more than the area of the principal peak
very slightly soluble in 2-propanol, practically insoluble in
in the chromatogram obtained with reference solution (a)
methylene chloride.
(0.1 per cent);
ID E N T IF IC A T IO N — unspecified impurities: for each impurity, no t m ore than the
A. Ultraviolet and visible absorption spectrophotometry area o f the principal peak in the chromatogram obtained
(2.2.25). w ith reference solution (a) (0.10 per cent);
1-370 Calcium Folinate 2016
— total: not more th a n twice the area o f the principal peak in — caldum (Ca; A r 40.08): 7.54 per cent to 8.14 per cent
the chromatogram obtained with reference solution (a) (anhydrous substance).
(0.2 per cent); It contains a variable quantity of water.
— disregard limir. 0.5 tim es the area o f the principal peak in
CHARACTERS
the chromatogram obtained with reference solution (a)
(0.05 per cent). A p p e a ra n c e
White or light yellow, amorphous or crystalline, hygroscopic
H e a v y m e ta ls (2.4.8) powder.
M axim um 15 ppm.
S o lu b ility
1.0 g complies with te s t C. Prepare the reference solution
Sparingly soluble in water, practically insoluble in acetone
using 1.5 m l. o f lead standard solution (10 ppm Pb) R.
and in ethanol (96 per cent).
Ir o n (2.4.9) T he amorphous form may produce supersaturated solutions
M axim um 10 ppm , determ ined on 10 m L of solution S. in water.
W a te r (2.5.12)
ID E N T IF IC A T IO N
4.0 p e r cent to 6.0 p e r cent, determined on 0.500 g.
First identification A , B, D
A SSA Y Second identification A , C, D
Dissolve 0.200 g in a m ixture o f 10 m l, of water R and
A. Specific optical rotation (see Tests).
40 m L of dilute sulfuric add R. T itrate with 0.1 M cerium
sulfate, d eterm ining th e end-point potentiometrically (2 .2 .2 0 ). B. Infrared absorption spectrophotometry (2.2.24).
1 m L o f 0.1 M cerium sulfate is equivalent to 10.45 m g of Preparation Discs.
Ci2HxoCaOioS2- Comparison caldum folinate CRS.
STO RA G E If the spectra obtained show differences, dissolve the
In an airtight container, protected from light substance to be examined and the reference substance
separately in the minim um volume of water R and add
IM P U R IT IE S dropwise suffident acetone R to produce a predpitate. Allow
Spedfied impurities A to stand for 15 min, collect the predpitate by centrifugation,
wash die predpitate with 2 small quantities o f acetone R and
HO
dry. Record new spectra using the residues.
C. Thin-layer chromatography (2.2.27).
OH
Test solution Dissolve 15 m g of the substance to be examined
A. benzene-1,4-diol (hydroquinone). in a 3 per cent V/V solution of ammonia R and dilute to
5 m L with the same solvent.
PhEur
Reference solution Dissolve 15 m g o f caldum folinate CRS in a
3 per cent V/V solution o f ammonia R and dilute to 5 m l,
with the same solvent.
★ ★ Plate cellulose for chromatography F2 5 4 R as the coating
Calcium Folinate ★ ★
substance.
(Ph Eur monograph 0978) *****
Mobile phase T he lower layer of a mixture of 1 volume of
isoamyl alcohol R and 10 volumes o f a 50 g/L solution of caric
add R previously adjusted to p H 8 with ammonia R.
Application 5 \iL.
Development Over a path o f 15 cm.
Drying In air.
Detection Examine in ultraviolet light at 254 nm.
Results T h e prindpal spot in the chromatogram obtained with
C20H 21CaN7O7^ H 2O 511.5 the test solution is sim ilar in position and size to the prindpal
spot in die chromatogram obtained with the reference
(anhydrous substance) solution.
D . It gives reaction (b) o f caldum (2.3.1).
A c tio n a n d u se Carry out the tests and the assay as rapidly as possible, protected
A ntidote to folic a d d antagonists. from actinic light.
P re p a r a tio n s
TESTS
C aldum Folinate Injection
S o lu tio n S
C ald u m Folinate Tablets Dissolve 1.25 g in carbon dioxide-free water R, heating at
PhEir------------------------------------ :-----------------------------------------
40 °C if necessary, and dilute to 50.0 m L with the same
solvent.
D E F IN IT IO N
A p p e a ra n c e o f so lu tio n
C aldum (2S)-2-[[4- [ [[ (6 RS) -2-amino-5-formyl-4-
Solution S is clear (2.2.1) and its absorbance (2.2.25) at
oxo 1,4,5,6,7,8-hexahydropteridin-6-
420 nm is not greater than 0.60. Use water R as the
yl] methyl] amino]benzoyl] amino]pentanedioate.
compensation liquid.
C o n te n t
p H (2.2.3)
— caldum folinate ^ o ^ j C a N y O y ) : 97.0 per cent to
6.8 to 8.0 for solution S.
102.0 per cent (anhydrous substance);
2016 Calcium Folinate 1-371
Specific optical rotation (2.2.7) phosphate R, previously adjusted to p H 7.8 with phosphoric
+ 14.4 to + 18.0 (anhydrous substance), determ ined on add R.
solution S. Flow rate 1 mL/min.
Acetone, ethanol and methanol Detection Spectrophotom eter at 280 nm.
Head-space gas chromatography (2.2.28) U se the standard Injection 10 pL o f the test solution and reference
additions method. solutions (b), (c), (d) and (e).
Test solution Dissolve 0.25 g o f the substance to be examined Run time 2.5 times the retention time o f folinate.
in water R and dilute to 10.0 m L with the same solvent.
System suitability: reference solution (e):
Reference solution Dilute 0.125 g of acetone R, 0.750 g of — resolution: minim um 2.2 between the peaks due to fo linate
anhydrous ethanol R and 0.125 g o f methanol R in water R and and impurity D .
dilute to 1000.0 m L with water R Limits:
Column: — impurity D: n o t m ore than the area o f the principal peak
— material: fused silica; in the chromatogram obtained with reference solution (c)
— sizer. I = 10 m , 0 = 0.32 mm; (1 per cent);
— stationary phase: styrene-divinylbenzene copolymer R. — impurities A , B, C, E, F, G: for each impurity, not more
Carrier gas nitrogen for chromatography R. than the area o f the principal peak in the chromatogram
Flow rate 4 mL/min. obtained with reference solution (b) (1 per cent);
Static head-space conditions that may be used: — sum of impurities other than D: not m ore than 2.5 times the
— equilibration temperature. 80 °C; area of the principal peak in the chromatogram obtained
— equilibration time: 20 min; with reference solution (b) (2.5 per cent);
— pressurisation time. 30 s. — disregard limit: the area of the principal peak in the
chromatogram obtained with reference solution (d)
Temperature:
(0.1 per cent).
Chlorides
Time Temperature
M aximum 0.5 per cent.
(min) CC)
Column 0 -6 125 -> 185 Dissolve 0.300 g in 50 m L of water R heating at 40 °C if
necessary. Add 10 m L o f 2 M nitric add and titrate with
6 - 15 185
0.005 M silver nitrate determining the end-point
Injection port 250 potentiometrically (2.2.20).
Detector 250 1 m L of 0.005 M silver nitrate is equivalent to 0.177 m g o f
Cl.
Heavy metals (2.4.8)
Detection Flam e ionisation. M aximum 50 ppm.
Irqecdon A t least 3 times. 1.0 g complies with test F. Prepare the reference solution
Limits: using 5 m L o f lead standard solution (10 ppm Pb) R.
— acetone: maximum 0.5 p er cent;
Platinum
— ethanol: maximum 3.0 per cent;
M aximum 20 ppm.
— methanol: m aximum 0.5 per c e n t
Atomic absorption spectrometry (2.2.23, Method II).
Related substances
Liquid chromatography (2.2.29). Test solution Dissolve 1.00 g in water R and dilute to
100.0 m L with the same solvent.
Test solution Dissolve 10.0 m g of the substance to be
examined in water R and dilute to 10.0 m L with the same Reference solutions Prepare the reference solutions using
solvent. platinum standard solution (30 ppm Pt) R, diluted as necessary
w ith a mixture o f 1 volume of nitric add R and 99 volumes of
Reference solution (a) Dissolve 10.0 m g o f calcium folinate CRS
water R.
in water R and dilute to 10.0 m L with the same solvent.
Source Platinum hollow-cathode lamp.
Reference solution (b) Dilute 1.0 m L o f reference solution (a)
to 100.0 m L with water R. Wavelength 265.9 nm.
Reference solution (c) Dissolve 10.0 mg o f formylfotic acid CRS Water (2.5.12)
(impurity D) in the mobile phase and dilute to 100.0 m L M aximum 17.0 per cent.
with the mobile phase. Dilute 1.0 m L o f this solution to Dissolve 0.100 g in a mixture o f 50 m L o f the titration
10.0 m L with water R. solvent and 15 m L o f formamide R. Stir for about 6 m in
Reference solution (d) Dilute 1.0 m l, o f reference solution (b) before titrating and use a suitable titrant th at does not
to 10.0 m L with water R. contain pyridine.
Reference solution (e) Dilute 5.0 m L of reference solution (c) Bacterial endotoxins (2.6.14)
to 10.0 m L with reference solution (b). Less than 0.5 IU/mg, if intended for use in the m anufacture
Column: o f parenteral preparations without a further appropriate
— size. I — 0.25 m , 0 = 4 m m ; procedure for the removal of bacterial endotoxins.
— stationary phase, octadecylsifyl silica gel for chromatography R ASSAY
(5 pm); Calcium
— temperature. 40 °C. Dissolve 0.400 g in 150 m L o f water R and dilute to 300 m L
Mobile phase Mix 220 m L o f methanol R and 780 m L o f a with the same solvent. Carry out the complexometric
solution containing 2.0 m L o f tetrabutylammonium hydroxide titration o f calcium (2.5.11).
solution (400 g/L) R and 2.2 g o f disodium hydrogen
1-372 Calcium Glucoheptonate 2016
IMPURITIES
Specified impurities A, B, C, D , E, F , G F. R = C H O : (25)-2-[[4-[[(2-am ino-4-oxo-l,4,7,8-
tetrahydropteridin-6-
yl) methyl] formylamino] benzoyl] amino] pentanedioic a d d
9 H COjH
(10-formyldihydrofolic ad d ),
G. R = H : (25) -2- [ [4- [ [(2-amino-4-oxo-1,4,7,8-
H,N c o 2h
tetrahydropteridin-6-
yl)methyl] amino] benzoyl] amino]pentanedioic a d d
(dihydrofolic add).
A. (2S)-2 [(4-aminobenzoyl) amino]pentanedioic acid,
PhEur
o H COjH
B. (2«S)-2-[[4- [ [[(6i?S)-2-amino-5-formyl-4-oxo-l,4,5,6,7,8-
hexahydropteridin-6-
Ca2* and epimer at C*
yl]methyi]formylamino]benzoyl]amino]pentanedioic a d d
(5,10-diformyltetrahydrofolic ad d ),
O H CO2H
Ci4H26C a016 490.4
o
Action and use
COjH
U sed in treatm ent o f caldum defidency.
h2n ^
x Xn ^ Tn^ ' *
2 H PhEur_____________________________________
DEFINITION
C . (2S)-2-[[4-[[(2-amino-4-oxo-1,4-dihydropteridin-6-
Mixture in variable proportions, o f caldum di(u-glycero-D-
yl)methyl]amino]benzoyl]amino]pentanedioic a d d (folic
^uZo-heptonate) and caldum di(D-^?yccrio-D-tii>-heptonate).
ad d ),
Content
98.0 per cent to 102.0 per cent o f caldum 2,3,4,5,6,7-
o h c o 2h -
hexahydroxyheptanoate (dried substance).
o r i T N '
CHARACTERS
'CC^H Appearance
White or very slightly yellow, amorphous powder,
iH0 hygroscopic.
H
Solubility
D. (26)-2-[[4-[ [(2-amino-4-oxo-1,4-dihydropteridin-6- Very soluble in water, practically insoluble in acetone and in
yl)methyl]formylamino]ben 2oyl]amino]pentanedioic a d d ethanol (96 per cent).
(10-formylfolic ad d ), IDENTIFICATION
A. Thin-layer chromatography (2.2.27).
Test solution Dissolve 20 m g of the substance to be examined
in 1 m L o f water R.
Reference solution (a) Dissolve 20 m g o f calcium
glucoheptonate CRS in 1 m l. of water R.
2016 Calcium Gluconate 1-373
TESTS S t x 50
S o lu tio n S S r —S t
T o 10.0 g add 90 m L of boiling distilled water Rand boil
with stirring, for not more than 10 s, until completely St = area of the peak due to oxalate in the
dissolved, then dilute to 100.0 m L with the same solvent. chromatogram obtained with die test solution;
A p p e a ra n c e o f so lu tio n Sr = area o f the peak due to oxalate in die
A t 60 °C, solution S is not more intensely coloured than chromatogram obtained with the reference solution.
reference solution B7 0- After cooling to 20 °C, it is not Limit.
more opalescent than reference suspension II (2.2.1). — oxalates: maximum 100 ppm.
p H (2.2.3) S u c ro se a n d re d u c in g su g a rs
6.4 to 8.3. Dissolve 0.5 g in a mixture of 2 m L o f hydrochloric add R1
Dissolve 1.0 g in 20 m L o f carbon dioxide-free water R, and 10 m L of water R. Boil for 5 m in, allow to cool, add
heating on a water-bath. 10 m L of sodium carbonate solution R and allow to stand for
O rg a n ic im p u ritie s a n d b o ric a c id 10 min. Dilute to 25 m L with water R and filter. T o 5 m L of
the filtrate add 2 m L o f cupri-tartaric solution R and boil for
Introduce 0.5 g into a porcelain dish previously rinsed with
1 min. Allow to stand for 2 m in. N o red precipitate is
sulfuric add R and placed in a bath of iced water. A dd 2 m L
formed.
of cooled sulfuric add R and mix. N o yellow or brown colour
develops. A dd 1 m L o f ckromotrope IIB solution R. A violet C h lo rid e s (2.4.4)
colour develops and does n o t become dark blue. M aximum 50 ppm.
T he solution is not more intensely coloured than that o f a T o 10 m L o f previously filtered solution S add 5 m L o f
mixture of 1 m L o f ckromotrope IIB solution R and 2 m L of water R.
cooled sulfuric add R.
P h o sp h a te s (2.4.11)
O x alates M aximum 100 ppm.
Liquid chromatography (2.2.29). D ilute 1 m L of solution S to 100 m L with water R.
Test solution Dissolve 1.00 g o f the substance to be examined S u lfates (2.4.13)
in water for chromatography R and dilute to 100.0 m L with
M aximum 50 ppm , determined on previously filtered
the same solvent.
solution S.
Reference solution Dissolve 1.00 g o f the substance to be Prepare the standard using a mixture o f 7.5 m L o f sulfate
examined in water for chromatography R, add 0.5 m L o f a standard solution (10 ppm SO J R and 7.5 m L of distilled
0.152 g/L solution of sodium oxalate R in water for water R.
chromatography R and dilute to 100.0 m L with the same
solvent. I ro n
M axim um 5 ppm.
Precolumn'.
— size. 1= 30 mm, 0 = 4 mm; Atomic absorption spectrometry (2.2.23, Method I).
— stationary phase, suitable strong anion-exchange resin Test solution Introduce 2.0 g into a 100 m L
(30-50 nm). polytetrafiuoroethylene beaker and add 5 m L o f nitric add R.
Columns 1 and 2: Boil, evaporating almost to dryness. Add 1 m L o f strong
— size. I = 0.25 m , 0 = 4 mm; hydrogen peroxide solution R and evaporate again almost to
— stationary phase, suitable strong anion-exchange resin dryness. Repeat the hydrogen peroxide treatm ent until a clear
(30-50 pm). solution is obtained. Using 2 m L o f nitric add R, transfer the
Anion-suppresser column Connected in series with the solution into a 25 m L volumetric flask. D ilute to 25.0 m L
precolumn and analytical columns and equipped with a with dilute hydrochloric add R. In the same m anner, prepare a
microm embrane that separates the mobile phase from the compensation solution using 0.65 g o f caldum chloride R1
instead of the substance to be examined.
suppressor regeneration solution, flowing countercurrent to
the mobile phase. Reference solutions Prepare the reference solutions from iron
Mobile phase Dissolve 0.212 g of anhydrous sodium carbonate R standard solution (20 ppm Fe) R, diluting with dilute
and 63 mg o f sodium hydrogen carbonate R in waterfor
hydrochloric add R.
chromatography R and dilute to 1000.0 m L with the same Source Iron hollow-cathode lamp.
solvent. Wavelength 248.3 nm.
Flow rate of the mobile phase 2 mL/min. Atomisation device Air-acetylene flame.
Suppressor regeneration solution 1.23 g/L solution o f sulfuric Carry out a basic correction using a deuterium lamp.
add R in water for chromatography R. M a g n e s iu m a n d a lk a li m e ta ls
Flow rate of the suppressor regeneration solution 4 m l Vmin. M aximum 0.4 per c e n t
Detection Conductance. T o 0.50 g add a mixture o f 1.0 m L o f dilute acetic add R and
Injection 50 |iL 10.0 m L o f water R and rapidly boil, whilst shaking, until
System suitability, reference solution: completely dissolved. T o the boiling solution add 5.0 m L of
— repeatability, maximum relative standard deviation of ammonium oxalate solution R and allow to stand for at least
2.0 per cent for die area o f the peak due to oxalate after 5 6 h. Filter through a sintered-glass filter ( 1. 6) (2.1.2) into a
injections. porcelain crucible. Carefully evaporate the filtrate to dryness
and ignite. T h e residue weighs no t more than 2 mg.
Inject 50 pL of each solution 3 times. Calculate the content
of oxalates in parts per million using die following H eav y m e ta ls (2.4.8)
expression: M axim um 10 ppm.
2016 Calcium Hydrogen Phosphate 1-377
Solubility Carry out the m easurem ent on 20.0 m L o f the test solution.
Practically insoluble in water and in ethanol (96 per cent). A dd at least 3 times 0.10 m L of the reference solution and
It dissolves in dilute hydrochloric acid and in dilute nitric carry out the m easurem ent after each addition. Calculate the
add. concentration o f fluorides using the calibration curve.
ID E N T IF IC A T IO N S u lfates
A. Dissolve with heating 0.1 g in 10 m L of dilute hydrochloric Maximum 0.5 per cent.
add R. A dd 2.5 m L o f dilute ammonia R l, shake and add Test solution Dissolve 0.5 g in a mixture o f 5 m L o f water R
5 m L of a 35 g/L solution of ammonium oxalate R. A white and 5 m L o f dilute hydrochloric add R and dilute to 100 m L
precipitate is produced. w ith water R. Filter if necessary. T o 20 m L o f this solution,
B. Dissolve 0.1 g in 5 m L o f dilute nitric add R, add 2 m L of add 1 m L of dilute hydrochloric add R and dilute to 50 m l.
ammonium molybdate solution R and heat at 70 °C for 2 min. with water R.
A yellow precipitate is produced. Reference solution T o 1.0 m L o f 0.005 M sulfuric add, add
C. It complies with the limits of the assay. 1 m L of dilute hydrochloric acid R -and dilute to 50 m L with
water R. Filter if necessary.
TESTS
T o the test solution and to the reference solution, add 2 m L
S o lu tio n S
of a 120 g/L solution o f barium chloride R and allow to stand
Dissolve 2.5 g in 20 m L of dilute hydrochloric acid R, filter if
for 10 min. Any opalescence in the test solution is not more
necessary and add dilute ammonia R l until a precipitate is
intense than that in the reference solution.
formed. A dd just sufficient dilute hydrochloric add R to
dissolve the precipitate and dilute to 50 m L with distilled A rse n ic (2.4.2, Method A)
water R. M aximum 10 ppm , determ ined on 2 m L o f solution s.
A cid -in so lu b le su b sta n c e s B a riu m
M aximum 0.2 per cent. T o 0.5 g, add 10 m L of water R and heat to boiling. While
Dissolve 5.0 g in 40 m L of water R, add 10 m L o f stirring, add 1 m L of hydrochloric acid R dropwise. Allow to
hydrochloric add R and heat to boiling for 5 min. Cool, then cool and filter if necessary. Add 2 m L of a 10 g/L solution of
collect the insoluble substances using ashless filter paper. dipotassium sulfate R and allow to stand for 10 m in.
N o turbidity is produced.
W ash with water R until turbidity is no longer produced
when silver nitrate solution R2 is added to the filtrate. Ignite at Ir o n (2.4.9)
600 ± 50 °C. T he residue weighs n o t more than 10 mg. M aximum 400 ppm.
C a rb o n a te s Dilute 0.5 m L o f solution s to 10 m L with water R.
Shake 0.5 g with 5 m L of carbon dioxide-free water R and add H eav y m e ta ls (2.4.8)
1 m L of hydrochloric add R. N o effervescence is produced. M aximum 40 ppm.
C h lo rid e s D ilute 10 m L o f solution s to 20 m L with water R. 12 m L of
M aximum 0.25 per cent. the solution complies with test A. Prepare the reference
Test solution Dissolve 0.20 g in a mixture of 20 m L o f water R solution using lead standard solution (1 ppm Pb) R.
and 13 m L o f dilute nitric add R by warming if necessary, L o ss on ig n itio n
dilute to 100 m L with water R and filter if necessary. 24.5 per cent to 26.5 per cent, determined on 1.000 g by
Use 50 m L of this solution. ignition to constant mass at 800-825 °c.
Reference solution T o 0.70 m L of 0.01 M hydrochloric add, add A SSAY
6 m l, of dilute nitric add R and dilute to 50 m L with water R.
Dissolve 0.4 g in 12 m L o f dilute hydrochloric acid R by
Add 1 m L o f silver nitrate solution R2 to the test solution and heating on a water bath if necessary and dilute to 200 m l.
to die reference solution and mix. After standing for 5 min with water R. T o 20.0 m L o f this solution add 25.0 m L of
protected from light, any opalescence in the test solution is 0.02 M sodium edetate, 50 m L of water R, 5 m L o f ammonium
n ot more intense than that in the reference solution. chloride buffer solution pH 10.7 R and about 25 m g o f mordant
F lu o rid e s black 11 triturate R. T itrate the excess o f sodium edetate with
M aximum 100 ppm. 0.02 M zinc sulfate. Carry o u t a blank titration.
Potentiom etry (2.2.36, Method II). 1 m L of 0.02 M sodium edetate is equivalent to 3.44 mg
Chelating solution Dissolve 45 g of cydohexylenedinitrilotetra- o f C a H P 0 4,2H 20 .
acetic acid R in 75 m L of sodium hydroxide solution R and F U N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S
dilute to 250 m L with water R. This section provides information on characteristics that are
Test solution Dissolve 1.000 g in 4 m L o f hydrochloric add R l, recognised as being relevant control parameters for one or more
add 20 m L o f chelating solution, 2.7 m L of glacial acetic functions of the substance when used as an excipient (see chapter
acid R and 2.8 g of sodium chloride R, adjust to p H 5-6 with 5.15). Some of the characteristics described in the Functionality-
sodium hydroxide solution R and dilute to 50.0 m L with related characteristics section may also be present in the mandatory
water R. part of the monograph since they also represent mandatory quality
Reference solution Dissolve 4.42 g o f sodium fluoride R, criteria. In such cases, a cross-reference to the tests described in the
previously dried at 300 °C for 12 h , in water R and dilute to mandatory part ừ included in the Functumality-related
1000.0 m L with the same solvent. Dilute 50.0 m L o f this characteristics section. Control of the characteristics can contribute
solution to 500.0 m L with total-tonic-strength-adjustment to the quality of a medicinal product by improving the consistency
buffer R (200 ppm F). of the manufacturing process and the performance of the medicinal
Indicator electrode Fluoride-selective. product during use. Where control methods are dted, they are
recognised as being suitable for the purpose, but other methods can
Reference electrode Silver-silver chloride. also be used Wherever results for a particular characteristic are
reported, the control 'method must be indicated
2016 Calcium Hydrogen Phosphate 1-379
The following characteristics may be relevant for calcium hydrogen A dd 1 m L o f silver nitrate solution R2 to the test solution and
phosphate dihydrate used as filler in tablets and capsules. to the reference solution and mix. A fter standing for 5 min
P a rtic le -siz e d is trib u tio n (2.9.31 or 2.9.38). protected from light, any opalescence in the test solution is
not m ore intense than that in the reference solution.
B u lk a n d ta p p e d d e n sity (2.9.34)
F lu o rid e s
P o w d e r flow (2.9.36) M axim um 100 ppm .
__________________________________________________________ PhEur
Potentiom etry (2.2.36> Method II).
Chdcamg solution Dissolve 45 g of cydohexylenedmitrilotetra-
acedc add R in 75 m L o f sodium hydroxide sdution R and
dilute to 250 m L with water R
Anhydrous Calcium Hydrogen ***** Test solution Dissolve 1.000 g in 4 m L o f hydrochloric add R l,
Phosphate ***** add 20 m L o f chelating solution, 2.7 m L o f glacial acetic
(Ph Eur monograph 0981) add R and 2.8 g o f sodium chloride R, adjust to p H 5-6 with
sodium hydroxide solution R and dilute to 50.0 m L with
C aH P04 136.1 7757-93-9 water R.
PhEur__________________________________________________________ Reference sdution Dissolve 4.42 g o i sodium fluoride R,
D E F IN IT IO N previously dried at 300 °C for 12 h , in water R and dilute to
C o n te n t 1000.0 m L with the same solvent. D ilute 50.0 m L of this
98.0 per cent to 103.0 per cent. solution to 500.0 m L with total-ionic-strength-adjustment
buffer R (200 ppm F).
CHA RACTERS
Indicator electrode Fluoride-selective.
A p p e a ra n c e
W hite or almost white, crystalline powder, or colourless Reference electrode Silver-silver chloride.
crystals. Carry o u t the m easurem ent on 20.0 m L of the test solution.
A dd at least 3 times 0.10 m L o f the reference solution and
S o lubility
carry out the m easurem ent after each addition. Calculate the
Practically insoluble in water and in ethanol (96 per cent).
It dissolves in dilute hydrochloric a d d and in dilute nitric concentration of fluorides using the calibration curve.
acid. S u lfa tes
M axim um 0.5 per cent.
ID E N T IF IC A T IO N
A. Dissolve with heating 0.1 g in 10 m L o f dilute hydrochloric Test solution Dissolve 0.5 g in a mixture of 5 m L o f water R
acid R. A dd 2.5 m L o f dUute ammonia R l, shake, and add and 5 m L o f dilute hydrochloric add R and dilute to 100 m L
5 m L of a 35 g/L solution o f ammonium oxalate R. A white w ith water R Filter if necessary. T o 20 m L of this solution,
add 1 m L o f dilute hydrochloric acid R and dilute to 50 m L
precipitate is produced.
with water R
B. Dissolve 0.1 g in 5 m L o f dilute nitric acid R, add 2 m L of
ammonium molybdate solution R and heat at 70 °C for 2 min. Reference solution T o 1.0 m L of 0.005 M sulfuric add, add
1 m L o f dilute hydrochloric add R a n d dilute to 50 m L with
A yellow precipitate is produced.
water R Filter if necessary.
C . It complies with the limits o f the assay.
T o the test solution and to the reference solution, add 2 m L
TESTS o f a 120 g/L solution o f barium chloride R and allow to stand
S o lu tio n S for 10 m in. Any opalescence in th e test solution is not more
Dissolve 2.5 g in 20 m L o f dilute hydrochloric add R, filter if intense than that in the reference solution.
necessary and add dilute ammonia R l until a precipitate is
A rse n ic (2.4.2, Method A)
formed. Add just sufficient dilute hydrochloric acid R to
M axim um 10 ppm , determined on 2 m L o f solution S.
dissolve'the precipitate and dilute to 50 m L with distilled
water R. B a riu m
T o 0.5 g, add 10 m L o f water R an d heat to boiling. While
A cid -in so lu b le su b s ta n c e s
stirring, add 1 m L of hydrochloric acid R dropwise. Allow to
Maximum 0.2 per cent.
cool and filter if necessary. Add 2 m L o f a 10 g/L solution o f
Dissolve 5.0 g in 40 m L o f water R, add 10 m L of dipotassium sulfate R and allow to stand for 10 min.
hydrochloric add R and heat to boiling for 5 min. Cool, then N o turbidity is produced.
collect the insoluble substances using ashless filter paper.
Ir o n (2.4.9)
W ash with water R until turbidity is no longer produced
M axim um 400 ppm .
w hen silver nitrate solution R2 is added. Ignite at
600 ± 50 °C. T he residue weighs not more th an 10 mg. Dilute 0.5 m L of solution S to 10 m L with water R.
C a rb o n a te s H eav y m e ta ls (2.4.8)
Shake 0.5 g with 5 m L o f carbon dioxide-free water R and add M axim um 40 ppm .
1 m L o f hydrochloric add R. N o effervescence is produced. Dilute 10 m L of solution S to 20 m L with water R. 12 m L o f
C h lo rid e s the solution complies with test A. Prepare the reference
solution using lead standard solution (1 ppm Pb) R.
M axim um 0.25 per cent.
Test solution Dissolve 0.20 g in a mixture o f 20 m L o f water R L oss o n ig n itio n
and 13 m L o f dilute nitric add R by warming if necessary, 6.6 p er cent to 8.5 p er cent, determ ined on 1.000 g to
dilute to 100 m L w ith water R and filter if necessary. constant mass at 800-825 °C.
U se 50 m L o f this solution.
Reference solution T o 0.70 m L of 0.01 M hydrochloric acid, add
6 m L of dilute nitric add R and dilute to 50 m L with water R.
1-380 Calcium Hydroxide 2016
A SSAY TESTS
Dissolve 0.4 g in 12 m L o f dilute hydrochbric acid R by M a tte r in so lu b le in h y d ro c h lo ric a c id
heatm g on a water b ath if necessary and dilute to 200 m L M axim um 0.5 per c e n t
with water R. T o 20.0 m L o f this solution add 25.0 m L of Dissolve 2.0 g in 30 m L of hydrochloric acid R. Boil the
0.02 M sodium edetate, 50 m L o f water R, 5 m L of ammonium solution and filter. Wash the residue w ith hot water R.
chloride buffer solution pH 10.7 R and about 25 mg o f mordant T h e residue weighs a m axim um of 10 mg.
black 11 triturate R. T itrate the excess o f sodium edetate with
C a rb o n a te s
0.02 M zinc sulfate. C an y out a blank titration.
M axim um 5.0 per cent o f C a C 0 3.
1 m L of 0.02 M sodium edetate is equivalent to 2.72 mg
Add 5.0 m L o f 1 M hydrochloric acid to the titrated solution
of C a H P 0 4.
obtained under Assay and titrate with 1 M sodium hydroxide
FU N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S using 0.5 m L of methyl orange solution R as indicator.
This section provides information on characteristics that are 1 m L of 1 M hydrochloric acid is equivalent to 50.05 mg
recognised as bang relevant control parameters for one or more o f C a C 0 3.
functions of the substance when used as an excipient (see chapter
C h lo rid e s (2.44)
5.15). Some of the characteristics described in the Functionality-
M aximum 330 ppm .
related characteristics section may also be present in the mandatory
part of the monograph since they also represent mandatory quality Dissolve 0.30 g in a mixture o f 2 m L o f nitric acid R and
criteria. In such cases, a cross-reference to the tests described in the 10 m L o f water R and dilute to 30 m L with water R.
mandatory part is included in the Functionality-related S u lfates (2.413)
characteristics section. Control of the characteristics can contribute M axim um 0.4 per cent.
to the quality of a medicinal product by improving the consistency Dissolve 0.15 g in a mixture o f 5 m L of dilute hydrochloric
of the manufacturing process and the performance of the medicinal acid R and 10 m L o f distilled water R and dilute to 60 m L
product during use. Where control methods are cited, they are with distilled water R.
recognised as being suitable for the purpose, but other methods can
A rse n ic (2.42, Method A)
also be used Wherever results for a particular characteristic are
M aximum 4 ppm.
reported, the control method must be indicated
Dissolve 0.50 g in 5 m L o f brominated hydrochloric arid R and
The following characteristics may be relevant for anhydrous
dilute to 50 m L with water R Use 25 m L o f this solution.
calcium hydrogen phosphate used as filler in tablets and capsules.
M a g n e s iu m a n d alkali m e ta ls
P a rd c le -s iz e d is trib u tio n (2.9.31)
Maximum 4.0 per cent calculated as sulfates.
or 2.9.38).
Dissolve 1.0 g in a mixture of 10 m L o f hydrochloric acid R
B u lk a n d ta p p e d d e n sity (2.9.34)
and 40 m L o f water R Boil and add 50 m L o f a 63 g/L
P o w d e r flow (2.9.36) solution o f oxalic acid R. Neutralise with ammonia R and
__________________________________________________________ PhEur dilute to 200 m L w ith water R Allow to stand for 1 h and
filter through a suitable filter. T o 100 m L of the filtrate, add
0.5 m L o f sul/uric acid R. Cautiously evaporate to dryness
and ignite. T he residue weighs a maximum of 20 mg.
★★*★★ H eav y m e ta ls (2.4.8)
Calcium Hydroxide ★ ★
M axim um 20 ppm.
(Ph. Eur. monograph 1078) * * * * *
Dissolve 1.0 g in 10 m L of hydrochloric acid R l and evaporate
Ca(OH)2 74.1 1305-62-0 to dryness on a water-bath. Dissolve the residue in 20 m L of
Preparation water R and filter. 12 m L of the filtrate complies with test A.
Calcium Hydroxide Solution Prepare the reference solution using lead standard solution
(1 ppm Pb) R.
PhEur_____________________________
ASSAY
D E F IN IT IO N
T o 1.500 g in a mortar, add 20-30 m L o f water R and
C o n te n t
0.5 m L o f phenolphthalein solution R. Titrate with 1 M
95.0 per cent to 100.5 per cent.
hydrochloric acid by triturating the substance until the red
CHARACTERS colour disappears. T he final solution is used in the tests for
A p p e a ra n c e carbonates.
W hite or almost white, fine powder. 1 m L of 1 M hydrochloric acid is equivalent to 37.05 mg
S o lubility o f C a(O H )2.
Practically insoluble in water. __________________________________________________________ PhEur
ID E N T IF IC A T IO N
A. T o 0.80 g in a mortar, add 10 m L o f water R and 0.5 m L
of phenolphthalein solution R and m k . T h e suspension turns
red. On addition of 17.5 m L of 1 M hydrochloric acid, the
suspension becomes colourless w ithout effervescing. T he red
colour occurs again when the mixture is triturated for 1 min.
O n addition of a further 6 m L o f 1 M hydrochloric acid and
triturating, the solution becomes colourless.
B. Dissolve about 0.1 g in dilute hydrochloric arid R and dilute
to 10 m L with water R. 5 m l, of the solution give
reaction (b) of calcium (2.3.1). •
2016 Calcium Lactate 1-381
A p p e a ra n c e o f so lu tio n
Calcium Lactate Pentahydrate ★* * * ★
Solution S is n o t more opalescent than reference ★ ★
suspension II (2.2.1) and n o t m ore intensely coloured than C ald u m Lactate *****
reference solution BY6 (2.2.2, Method II). (Ph. Eur. monograph 0468)
A cid ity o r alk a lin ity
T o 10 m L o f solution S add 0.1 m L o f phenolphthalein
solution R and 0.5 m L of 0.01 M hydrochloric acid. Ca2* 'X and enantiomer , 5 H20
T he solution is colourless. N o t more than 2.0 m L of 0.01 M H OH
Test solution Dissolve 10.0 mg of the substance to be Mobile phase Dissolve 9.72 g of sodium dihydrogen phosphate R
examined in water R and dilute to 10.0 m L w ith the same in 890 m l, of water R and adjust to p H 5.0 with sodium
solvent. hydroxide solution R; add 100 m L o f 2-propanol R and 10 m L
Reference solution (a) Dissolve 10.0 mg of calcium folinate CRS of acetonitrUe R.
in water R and dilute to 10.0 m L with the sam e solvent. Flow rate 1 mL/min.
Reference solution (b) Dilute 1.0 m L of reference solution (a) Detection Spectrophotometer at 286 nm.
to 100.0 m L with water R. Injection 10 pL.
Reference solution (c) Dissolve 10.0 m g o f forrnylfoHc acid CRS Retention times Levofolinate = about 9 min;
in the mobile phase and dilute to 100.0 m L with the mobile im purity H = about 19 min.
phase. Dilute 1.0 m L of this solution to 10.0 m L with System suitability.
water R. — resolution: m in im u m of 5.0 between the peaks due to
Reference solution (d) Dilute 1.0 m L o f reference solution (b) levofolinate and to impurity H in the chromatogram
to 20.0 m L with water R. obtained with reference solution (a). T he sum o f the
Reference solution (e) Dilute 5.0 m L o f reference solution (c) areas o f the 2 peaks is 100 per cent. T he peak area of
to 10.0 m L with reference solution (b). impurity H is 48 per cent to 52 per cent. In the
Column: chromatogram obtained with reference solution (b) 2
— size: I = 0.25 m, 0 = 4 mm; clearly visible peaks are obtained.
— stationary phase: octadecylsilyl silica gelfor chromatography R Limit:
(5 pm); — impurity H: maximum 0.5 per cent.
— temperature: 40 °C. C h lo rid e s
Mobile phase Mix 220 m L o f methanol R and 780 m L of a M axim u m 0.5 per cent.
solution containing 2.0 m L o f tetrabutylamrnonium hydroxide Dissolve 0.300 g in 50 m L of water R heating at 40 °C if
solution (400 g/L) R and 2.2 g o f disodium hydrogen necessary. Add 10 m L o f 2 M nitric acid and titrate with
phosphate R previously adjusted to p H 7.8 w ith phosphoric 0.005 M silver nitrate determining the end-point
acid R. If necessary adjust the concentration o f methanol R to potentiometrically (2.2.20).
achieve the prescribed resolution. 1 m L o f 0.005 M silver nitrate is equivalent to 0.177 mg o f
Flow rate 1 mL/min. Cl.
Detection Spectrophotometer at 280 nm. P la tin u m
Injection 10 pL. M axim um 10 ppm .
Run time 2.5 times the retention time of the principal peak in Atomic absorption spectrometry (2.2.23, Method II).
the chromatogram obtained with the test solution. Test solution Dissolve 1.0 g in water R and dilute to 100.0 m L
System suitability, reference solution (e): with the same solvent.
— resolution: minim um o f 2.2 between the peaks due to Reference solutions Prepare the reference solutions losing
folinate and to impurity D. platinum standard sdution (30 ppm Pt) R, diluted as necessary
Limits: w ith a m ixture o f 1 volume of nitric acid R and 99 volumes of
— impurity D: n o t more than 0.8 times the area o f the water R.
principal peak in the chromatogram obtained with
Source Platinum hollow-cathode lamp.
reference solution (c) (0.8 per cent);
— any other impurity, not more than 0.8 times the area o f the
Wavelength 265.9 nm.
principal peak in the chromatogram obtained with H eav y m e ta ls (2.4.8)
reference solution (b) (0.8 per cent); M axim um 50 ppm .
— sum of other impurities: not more than twice the area o f the 1.0 g complies with test F. Prepare the reference solution
principal peak in the chromatogram obtained with using 5 m L of lead standard solution (10 ppm Pb) R.
reference solution (b) (2.0 per cent); W a te r (2.5.12)
— disregard Umif. area of the principal peak in the 10.0 per cent to 17.0 per cent, determined on 0.200 g
chromatogram obtained with reference solution (d) (ground to a very fine powder). Stir the substance to be
(0.05 per cent). examined in the titration solvent for about 15 m in before
Im p u rity H titrating and use iodosidfurous reagent R as titranL
Liquid chromatography (2.2.29): use the normalisation B a c te ria l en d o to x in s (2.6.14)
procedure. Less than 0.5 IU/mg, if intended for use in the m anufacture
Test solution Dissolve 50.0 mg of the substance to be o f parenteral preparations without a further appropriate
examined in water R and dilute to 100.0 m L with the same procedure for the removal of bacterial endotoxins.
solvent.
A SSA Y
Reference solution (a) Dissolve 10.0 mg of caldum folinate CRS C a lc iu m
in water R and dilute to 20.0 m L with the same solvent.
Dissolve 0.400 g in 150 m L o f water R and dilute to 300 m L
Reference solution (b) Dilute 1.0 m L o f reference solution (a) w ith the same solvent. Carry out the complexometric
to 100.0 m L with water R. titration o f calcium (2.5.11).
Column: 1 m L of 0.1 M sodium edetate is equivalent to 4.008 mg o f
— size: I = 0.15 m , 0 = 4 mm; Ca.
— stationary phase: human albumin coated silica gel for
C a lc iu m fo lin ate
chromatography R (5 pm);
Liquid chromatography (2.2.29) as described in the test for
— temperature: 40 °C.
related substances.
1-386 Calcium Levulinate 2016
STORAGE H,N
In a n airtight container, protected from light. If the substance
is sterile, store in a sterile, airtight, tam per-proof container.
F. R = C H O : (2S)-2-[[4-[[(2-am ino-4-oxo-l,4,7,8-
IM P U R IT IE S tetrahydropteridin-6-
yl)methyl]formylamino]benzoyl]amino]pentanedioic a d d
(10-formyldihydrofolic ad d ),
G . R = H: (2S)-2-[[4-[[(2-am ino-4-oxo-l,4,7,8-
tetrahydropteridin-6-
H,N C02H yl)methyl] amino]benzoyl] amino] pentanedioic a d d
(dihydrofolic ad d ),
A. (2S)-2-[(4-aminobenzoyl) amino] pentanedioic acid,
O h co 2h
o H C02H
co 2h
co 2h
h2n n n
H H
h2n n n
H H
H . (2S)-2-[[4-[[[(6i?)-2-am ino-5-fonnyl-4-oxo-l,4,5,6,7,8-
B. (25)-2-[[4-[[[(6i?)-2-am ino-5-form yl-4-oxo-l,4,5,6,7,8- hexahydropteridin-6-
hexahydropteridin-6- yl] methyl] amino]benzoyl]amino]pentanedioic ad d .
yl] methyl]fonnylamino]benzoyi] amino]pentanedioic acid PtiEur
(5,10-difonnyltetrahydrofolic ad d ),
9 H c o 2h
★ ★
Calcium Levulinate Dihydrate ★ ★
(Ph. Eur. monograph 1296) * * * * *
h2n n n
C02*
Ca2* h3CY ^ , 2 H20
C . (2S)-2- [[4- [[(2-amino-4-oxo-l,4-dihydropteridin-6-
yl)methyl] amino]benzoyl]amino]pentanedioic a d d (folic
a d d ),
C io H 14C a 0 6 ^ H 20 306.3 5743-49-7
A ctio n a n d u se
Source o f caldum .
0 f
C02H PhEur_____________________________
CHO D E F IN IT IO N
h2n n n
2 H C ald u m di(4-oxopentanoate) dihydrate.
C o n te n t
D . (2S)-2-[[4-[[(2-amm o-4-oxo-1,4-dihydropteridin-6- 98.0 per cent to 101.0 per cent (dried substance).
yl)methyi]fonnylamino]benzoyl] amino]pentanedioic a d d
(10-fonnylfolic ad d ), CHARACTERS
A p p e a ra n c e
W hite or almost white, crystalline powder.
»XT Solubility
F red y soluble in water, very slightly soluble in ethanol
(96 per cent), practically insoluble in methylene chloride.
h2n n n
H H ID E N T IF IC A T IO N
First identificatidn A , D, E.
E. 4-[[[(65)-2-am ino-5-form yl-4-oxo-l,4,5,6,7,8- Second identification B, C, D, E.
hexahydropteridin-6-yl]methyl]amino]benzoic a d d
A. Infrared absorption spectrophotom etry (2.2.24).
(5-fonnyitetrahydropteroic a d d ),
Comparison calcium levulinate dihydrate CRS.
B. Thin-layer chromatography (2.2.27).
Test solution Dissolve 60 mg o f the substance to be examined
in water R and dilute to 1 m L with .the same solvent.
2016 Calcium Pantothenate 1-387
Reference solution Dissolve 60 mg of calcium leuulinate dryness on a water-bath and ignite to constant mass at
dihydrate CRS in water R and dilute to 1 m L with the same 600 ± 50 °C. T he residue wdghs a maximum of 5.0 mg.
solvent. H eav y m e ta ls (2.4.8)
Plate TLC silica gel plate R. M aximum 10 ppm .
Mobile phase concentrated ammonia R, ethyl acetate R, water R, 12 m L o f solution S complies with test A. Prepare the
ethanol (96 per cent) R (10:10:30:50 VIVIVIV). reference solution using lead standard solution (1 ppm Pb) R.
Application 10 |iL. L oss o n d ry in g (2.2.32)
Development Over a path of 10 cm. 11.0 per cent to 12.5 per cent, determined on 0.200 g by
Drying A t 100-105 °C for 20 min and allow to cool. drying at 105 °C.
Detection Spray with a 30 g/L solution of potassium P y ro g e n s (2.6.8)
permanganate R. D ry in a current of warm air for about If intended for use in the manufacture of parenteral
5 min or until the spots become yellow. Examine in daylight. preparations without a further appropriate procedure for the
Results T h e prindpal spot in the chromatogram obtained with removal o f pyrogens, it complies with the test for pyrogens.
the test solution is similar in position, colour and size to the Inject per kilogram of the rabbit’s mass 4 m L of a solution
principal spot in the chromatogram obtained with the containing per millilitre 50 mg of the substance to be
examined.
reference solution.
C. T o 1 m L of solution S (see Tests), add 20 m L of a ASSAY
2.5 g/L solution o f dinitrophenyütydrazine R in dilute Dissolve 0.240 g in 50 m L of water R. C any out the
hydrochloric acid R. Allow to stand for 15 min. Filter, wash complexometric titration o f caldum (2.5.11).
the precipitate with water R. Dry the precipitate in an oven at 1 m L of 0.1 M sodium edetate is equivalent to 27.03 mg
100-105 °C. T he melting point (2.2.14) is 203 °C to 210 °C. o f CioH14C a 0 6.
D. It gives reaction (b) o f caldum (2.3.1). STORA GE
E. Loss on drying (see Tests). Protected from light.
TESTS __________________________________________________________PhEur
S o lu tio n S
Dissolve 10.0 g in carbon dioxide-free water R prepared from
distilled water R and dilute to 100.0 m L with the same
solvent. ***
Calcium Pantothenate ★ ★
A p p e a ra n c e o f so lu tio n ★ ★
Solution S is d ear (2.2.1) and not more intensely coloured (Ph. Eur. monograph 0470) *****
than reference solution Y6 (2.2.2, Method II).
p H (2.2.3) h3c c H30
6.8 to 7.8 for solution S. Cat1* H0- V n
O x id isab le su b sta n c e s / oh h
T o 1 m L o f solution S, add 10 m L o f water R, 1 m L o f dilute
sulfuric add R and 0.25 m L o f a 3.0 g/L solution o f potassium
permanganate R. Mix. After 5 min, the violet colour o f the C x8H 32CaN 2O10 476.5 137-08-6
mixture is still visible.
A c tio n a n d u se
S u c ro se a n d re d u c in g su g a rs
C om ponent of vitamin B.
T o 5 m L o f solution S add 2 m L of hydrochloric add R l and
dilute to 10 m L with water R. H eat to boiling for 5 m in and PhE ir______________________
allow to .cool. Add 10 m L of sodium carbonate solution R.
D E F IN IT IO N
Allow to stand for 5 min, dilute to 25 m L with water R and
filter. T o 5 m L o f die filtrate add 2 m L o f cupri-tartaric C aldum pantothenate contains not less than 98.0 per cent
solution R and heat to boiling for 1 min. N o red predpitate is and not more than the equivalent of 101.0 per cent of
formed. caldum bis[3-[[(21?)-2,4-dihydroxy-3,3-
dimethyibutanoyl]amino]propanoate], calculated with
C h lo rid e s (2.4.4) reference to the dried substance.
M aximum 50 ppm.
CHARACTERS
Dilute 10 m L o f solution S to 15 m L with water R.
A white or almost white powder, slightly hygroscopic, fredy
S u lfates (2.4.13) soluble in water, slightly soluble in alcohol.
M axim um 200 ppm.
ID E N T IF IC A T IO N
Dilute 7.5 m L o f solution S to 15 m L with distilled water R.
A. Specific optical rotation (see Tests).
M a g n e s iu m a n d alk ali m e ta ls
B. Examine the chromatograms obtained in the test for
M axim um 1.0 per c e n t
3-aminopropionic ad d . T he principal spot in the
T o 10 m L o f solution S, add 80 m L of water R, 10 m L of chromatogram obtained with test solution (b) is similar in
ammonium chloride solution R and 1 m L o f ammonia R. H eat position, colour and size to the prindpal spot in the
to boiling. T o the boiling solution, add dropwise 50 m L of chromatogram obtained with reference solution (a).
warm ammonium oxalate sdution R. Allow to stand for 4 h,
C . T o 1 m L o f solution S (see Tests) add 1 m L o f dilute
then dilute to 200 m L with water R and filter. T o 100 m L of
sodium hydroxide sdution R and 0.1 m L o f copper sulfate
the filtrate, add 0.5 m L of sulfuric add R. Evaporate to
solution R. A blue colour develops.
D . It gives reaction (a) of caldum (2.3.1).
1-388 Calcium Phosphate 2016
TESTS ★ ★
S o lu tio n S
Calcium Phosphate ★ ★
Dissolve 2.50 g in carbon dioxide-free water R and dilute to Tribasic C ald u m Phosphate *****
50.0 m L w ith the same solvent. (Ph. Eur. monograph 1052)
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and colourless (2.2.2, Method II). A c tio n a n d u se
Exdpient.
p H (2.2.3)
T he p H o f solution S is 6.8 to 8.0. P re p a r a tio n
C aldum and Ergocalciferol Tablets
S p ecific o p tic a l ro ta tio n (2.2.7)
+ 25.5 to + 27.5, determined on solution S and calculated C aldum Phosphate for H om oeopathic Preparations
with reference to the dried substance. Chewable C ald u m and Ergocalciferol Tablets
3-A m in o p ro p io n ic a c id PhEui.
Examine by thin-layer chromatography (2.2.27), using silica
gel G R as the coating substance. D E F IN IT IO N
M ixture o f caldum phosphates.
Test solution (a) Dissolve 0.2 g o f the substance to be
examined in water R and dilute to 5 m L with the same C o n te n t
solvent. 35.0 per cent to 40.0 per cent o f C a (Ar 40.08).
Test solution (b) Dilute 1 m L o f test solution (a) to 10 m L CHARACTERS
w ith water R. A p p e a ra n c e
Reference solution (a) Dissolve 20 m g o f calcium W hite or almost white powder.
pantothenate CRS in water R and dilute to 5 m L with the S o lu b ility
same solvent Practically insoluble in water. It dissolves in dilute
Reference solution (b) Dissolve 10 mg o f 3-aminopropionic hydrochloric a d d and in dilute nitric ad d .
acid R in water R and dilute to 50 mT. with the same solvent.
ID E N T IF IC A T IO N
Apply separately to th e plate 5 pL o f each solution. Develop A. Dissolve 0.1 g in 5 m L o f a 25 p er cent VIV solution of
over a path o f 12 cm using a mixture of 35 volumes of nitric acid R. T he solution gives reaction (b) o f phosphates
water R ahd 65 volumes of ethanol R. D ry the plate in a (2.3.1).
current of air and spray with nmhydrin solution R l. H eat at
B. It gives reaction (b) o f caldum (2.3.1). Filter before
110 °C for 10 min. Any spot corresponding to
adding potassium ferrocyanide solution R.
3-am inopropionic a d d in the chrom atogram obtained with
test solution (a) is n o t more intense than the spot in the C. It complies with the limits of the assay.
chrom atogram obtained with reference solution (b) TESTS
(0.5 per cent). S o lu tio n S
C h lo rid e s (2.4.4) Dissolve 2.50 g in 20 mT. o f dilute hydrochloric add R. If the
5 m L o f solution S diluted to 15 m L with water R complies solution is n o t clear, filter it. Add dilute ammonia R l dropwise
with the limit test for chlorides (200 ppm ). until a predpitate is formed. Dissolve the predpitate by
H eav y m e ta ls (2.48) adding dilute hydrochloric add R and dilute to 50 m L w ith
12 m L o f solution S complies with test A for heavy metals
distilled water R.
(20 ppm ). Prepare the reference solution using lead standard C h lo rid e s (2.44)
solution (1 ppm Fb) R. M aximum 0.15 per cent.
L oss o n d ry in g (2.2.32) Dissolve 0.22 g in a mixture o f 1 m L o f nitric add R and
N ot m ore than 3.0 per cent, determ ined on 1.000 g by 10 m L of water R and dilute to 100 m L with water R.
drying in an oven at 105 °C. F lu o rid e s
A SSA Y M axim um 75 ppm .
Dissolve 0.180 g in 50 m L o f anhydrous acetic acid R. T itrate Potentiom etry (2.2.36, Method II).
with 0.1 M perchloric acid determ in in g the end-point Test solution Dissolve 0.250 g in 0.1 M hydrochloric add, add
potentiometrically (2.2.20). 5.0 m L of fluoride standard solution (1 ppm F) R and dilute to
1 m L o f 0.1 M perchloric add is equivalent to 23.83 m g o f 50.0 mT. with 0.1 M hydrochloric acid. T o 20.0 m L o f this
Ci8H32C aN 2O 10. solution add 20.0 mT. o f total-ionic-strength-adjustment buffer R
and 3 m L o f an 82 g/L solution o f anhydrous sodium
STORAGE
acetate R. Adjust to p H 5.2 with ammonia R and dilute to
Store in an airtight container.
50.0 m L with distilled water R.
PhEtr Reference solution Fluoride standard solution (10 ppm F) R.
Indicator electrode Fluoride-selective.
Reference electrode Silver-silver chloride.
Carry out the measurem ents on the test solution, th en add at
least 3 quantities, each o f 0.5 m L, o f the reference solution,
carrying out a m easurem ent after each addition. Calculate
the concentration o f fluorides using the calibration curve,
taking into account the addition o f fluoride to the test
solution.
2016 Calcium Polystyrene Sulfonate 1-389
S u lfates (.2.4.13)
M axim um 0.5 per c e n t
Calcium Polystyrene Sulfonate
Calcium Polystyrene Sulphonate
D ilute 1 m L o f solution S to 25 m L with distilled water R.
A rsen ic (2.4.2, Method A) A c tio n a n d u se
M axim um 4 ppm , determined on 5 m L of solution S. U sed in the treatm ent of hyperkalaemia.
Ir o n (2.4.9)
D E F IN IT IO N
M axim um 400 ppm.
Calcium Polystyrene Sulfonate is a cation-exchange resin
Dilute 0.5 m L of solution S to 10 m L with water R.
prepared in the calcium form containing not less than 6.5%
H eav y m e ta ls (2.4.8) w/w and n o t m ore than 9.5% w/w of calcium, calculated with
M axim um 30 ppm . reference to die dried substance. Each g exchanges not less
D ilute 13 m L o f solution S to 20 m L with water R. 12 m L of than 1.3 m E q and not more than 2.0 m E q of potassium,
the solution complies with test A. Prepare the reference calculated with reference to the dried substance.
solution using lead standard solution (1 ppm Pb) R. C H A R A C T E R IS T IC S
A cid -in so h ib le m a tte r A cream to light brown, fine powder.
M aximum 0.2 per cent. Practically insoluble in water and in ethanol (96%).
Dissolve 5.0 g in a mixture o f 10 m L o f hydrochloric acid R
ID E N T IF IC A T IO N
and 30 m L of water R. Filter, wash the residue with water R
A. T h e infrared absorption spectrum, Appendix II A, is
and dry to constant mass at 100-105 °C. T h e residue weighs
concordant with the reference spectrum of calcium polystyrene
a maximum o f 10 mg.
sulfonate (RS 037).
L oss o n ig n itio n
B. Yields reaction C characteristic o f calcium salts,
M aximum 8.0 per cent, determined on 1.000 g by ignition at
Appendix VI.
800 ± 50 °C for 30 min.
TESTS
A SSAY
P a rtic le size
Dissolve 0.200 g in a mixture of 1 m L of hydrochloric acid R1
N o t more than 1% w/w is retained on a 150-pm sieve,
and 5 m L o f water R. Add 25.0 m L of 0.1 M sodium edetate
Appendix X V IIB . Use 20 g and sieve for 5 minutes.
and dilute to 200 m L with water R. Adjust to about p H 10
with concentrated ammonia R. Add 10 m L o f ammonium P o ta s s iu m
chloride buffer solution pH 10.0 R and a few milligrams of N o t more than 0.1% of K when determined by atomic
mordant black 11 triturate R. Titrate the excess sodium edetate emission spectrophotometry, Appendix II D , measuring at
with 0.1 M zinc sulfate until the colour changes from blue to 766.5 nm and using a solution prepared in the following
violet. m anner. T o 1.1 g of the substance being examined add
5 m L of hydrochloric add, heat to boiling, cool and add
1 m L of 0.1 M sodium edetate is equivalent to 4.008 m g of
10 m L o f water. Filter, wash the filter and residue with water
Ca.
and dilute the filtrate and washings to 25 m L with water. Use
FU N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S potassium standard solution (100 ppm K), suitably diluted with
This section provides mformaxion on characteristics that are water, to prepare the standard solutions.
recognised as being relevant control parameters for one or more S o d iu m
functions of the substance when used as an excipient (see chapter N o t more than 0.1% of N a when determined by atomic
5.15). This section is a non-mandatory pan of the monograph emission spectrophotometry, Appendix II D , measuring at
and it is not necessary to verify the characteristics to demonstrate 589.0 nm and using a solution prepared in the following
compliance. Control of these characteristics can however contribute m anner. T o 1.1 g of the substance being examined add
to the quality of a medicinal product by improving the consistency 5 m L of hydrochloric add, heat to boiling, cool and add
of the manufacturing process and the performance of the medicinal 10 m L of water. Filter, wash the filter and residue with water
product during use. Where control methods are cited, they are and dilute the filtrate and washings to 25 m L with water. Use
recognised as being suitable for the purpose, but other methods can sodium solution (200 ppm Na), suitably diluted with water, to
also be used Wherever results for a particular characteristic are prepare the standard solutions.
reported, the control method must be indicated
A rsen ic
The following characteristics may be relevant for calcium
1 g complies with the limit test for arsenic, Appendix VII
phosphate is used as a filler in tablets and capsules.
(1 ppm ).
P a r d d e - s iz e d is trib u tio n (2.931 or 2.938).
H eav y m e ta ls
B u lk a n d ta p p e d d e n sity (2.934) H eat 4 g until charred, cool, add 4 m L o f lead-free nitric acid
P o w d e r flow (2.9.36) and 0.5 m L o f sulfuric add drop wise and heat cautiously
PhEur until white fumes are no longer evolved. Ignite in a muffle
furnace at 500° to 600° until a white residue is obtained.
Cool, add 4 m L o f hydrochloric add and dilute to 20 mL.
T h e resulting solution complies with limit test A for heavy
metals, Appendix VII. U se 2 m L o f lead standard solution
(lOppmPb) to prepare the standard (10 ppm).
S ty re n e
Carry out the m ethod for liquid chromatography,
Appendix ID D , using the following solutions.
1-390 Calcium Stearate 2016
C h lo rid es (2.4.4) and titrate with 0.1 M zinc sulfate until the colour changes
Maximum 0.1 p er c e n t from blue to violet Carry out a blank titration.
Dilute 0.5 m L o f solution S to 15 m L with water R. 1 m L o f 0.1 M sodium edetate is equivalent to 4.008 mg of
S u lfates (2.4.13) Ca.
Maximum 0.3 per c e n t C o m p o sitio n o f fatty acids
Dilute 0.5 m L of solution S to 15 m L with distilled water R. Gas chromatography (2.2.28): use the normalisation
procedure.
C a d m iu m
M aximum 3 ppm . Test solution In a conical flask fitted with a reflux condenser,
dissolve 0.10 g of the substance to be examined in 5 m L of
Atomic absorption spectrometry {2.2.23, Method II).
boron trifluoride-methanol solution R. Boil under a reflux
Test solution Place 50.0 mg in a polytetrafluoroethylene condenser for 10 min. Add 4 m L o f heptane R through the
digestion bom b and add 0.5 m L o f a mixture o f 1 volume of condenser. Boil under a reflux condenser for 10 min. Allow
hydrochloric add R and 5 volumes of cadmium- and lead-free to cool. Add 20 m L of saturated sodium chloride solution R.
nitric add R. Allow to digest at 170 °C for 5 h. Allow to cool. Shake and allow the layers to separate. Remove about 2 m L
Dissolve the residue in water R and dilute to 5.0 m L with the o f the organic layer and dry over 0.2 g of anhydrous sodium
same solvent. sulfate R. Dilute 1.0 m L of the solution to 10.0 m L with
Reference solutions Prepare the reference solutions using heptane R.
cadmium standard solution (10 ppm Cd) R, diluted if necessary Reference solution Prepare the reference solution in the same
with a 1 per cent VIV solution of hydrochloric add R. m anner as the test solution using 50.0 mg oipalmitic
Source Cadm ium Hollow-cathode lamp. add CRS and 50.0 mg o f stearic add CRS instead of calcium
Wavelength 228J3 nm . stearate.
Atomisation device Graphite furnace. Cdumn:
— material: fused silica;
L ead
— size. I = 30 m , 0 = 0.32 mm;
M aximum 10 ppm .
— stationary phase: macrogol 20 000 R (film thickness
Atomic absorption spectrometry (2.2. 23, Method II). 0.5 |im).
Test solution Use the solution described in the test for Carrier gas helium for chromatography R.
cadmium.
Flow rate 2.4 mlVmin.
Reference solutions Prepare the reference solutions using lead Temperature.
standard solution (10 ppm Pb) R, diluted if necessary with
water R.
Source Lead hollow-cathode lamp. Time Temperature
(min) <°C)
Wavelength 283.3 nm ; 217.0 nm may be used depending on 0 -2 70
Column
the apparatus.
2 -3 6 70 -> 240
Atomisation device Graphite furnace.
36-41 240
N ickel
M aximum 5 ppm . Injection port 220 ..
Atomic absorption spectrometry (2.2.23, Method II). Detector 260
Test solution Use the solution described in the test for
cadmium.
Reference solutions Prepare the reference solutions using nickel Detection Flame ionisation.
standard solution (10 ppm Ni) R, diluted if necessary with Injection 1 ¿iL.
water R. Relative retention W ith reference to methyl stearate: methyl
Source Nickel hollow-cathode lamp. palmitate = about 0.9.
Wavelength 232.0 nm. System suitability: reference solution:
— resolution: minimum 5.0 between the peaks due to methyl
Atomisation device Graphite furnace.
palmitate and methyl stearate.
L oss o n d ry in g (2.2.32)
Calculate the content of palmitic acid and stearic acid.
M aximum 6.0 p er cent, determined on 1.000 g by drying in
Disregard the peak due to the solvent.
an oven at 105 °C.
F U N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S
M ic ro b ia l c o n ta m in a tio n
TA M C: acceptance criterion 103 C FU/g (2.6.12).
This section provides information on characteristics that are
recognised as being relevant control parameters for one or more
TY M C: acceptance criterion 102 C FU /g (2.6.12).
Junctions of the substance when used as an excipient (see chapter
Absence of Escherichia coli (2.6.13). 5.15). This section is a non-mandatory part of the monograph
Absence o f Salmonella (2.6.13). and it is not necessary to verify the characteristics to demonstrate
ASSA Y
compliance. Control of these characteristics can however contribute
to the quality of a medicinal product by improving the consistency
C a lc iu m
of the manufacturijig process and the performance of the medicinal
T o 0.500 g in a 250 m L conical flask add 50 m L o f a
product during use. Where control methods are cited, they are
mixture of equal volumes of anhydrous ethanol R and
recognised as bring suitable for the purpose, but other methods can
butanol R, 5 m L o f concentrated ammonia R, 3 m L of
also be used Wherever results for a particular characteristic are
ammonium chloride buffer solution pH 10.0 R, 30.0 m L of
reported, the control method must be indicated.
0.1 M sodium edetate and 15 mg o f mordant black 11
triturate R. H eat to 45-50 °C until the solution is clear. Cool
1-392 Calcium Sulfate 2016
P o w d e r flow (2.9.36)
Test solution Dissolve 2.50 g of the substance to be examined
__________________________________________________________________________________________ P hEur
in heptane R and dilute to 25.0 m L with the same solvent.
Reference solution (a) Dilute 1.0 m l. of the test solution to
100.0 m L with heptane R.
Reference solution (b) Dilute 10.0 m L of reference solution (a)
to 20.0 m L with heptane R.
Natural Camphor ;* % Reference solution (c) Dissolve 0.50 g of bomeol R in heptane R
*★ and dilute to 25.0 m L with the same solvent. D ilute 5.0 m L
(o-Camphor, Ph Eur monograph 1400) *
of the solution to 50.0 m L with heptane R.
CH3 Reference solution (d) Dissolve 50 m g of linalol R and 50 mg
o f bomyl acetate R in heptane R and dilute to 100.0 m L with
the same solvent.
Column:
H — size. I = 30 m , 0 = 0.25 mm,
— stationary phase: macrogol 20 000 R (0.25 pm).
Carrier gas helium for chromatography R.
C 10H 16O 152.2 464-49-3
Split ratio 1:70.
Ph E tr _____________________ I ; -------------------------------------------------------------------------------------------------------
Flow rate 45 cm/s.
D E F IN IT IO N Temperature:
(li?,4/?)-l ,7,7-Trimethylbicyclo [2.2. l]heptan-2-one.
CHARACTERS Time Temperature
A p p e a ra n c e (min) (°C)
W hite or almost white, crystalline powder or friable, Column 0-10 50
crystalline masses.
10-35 50 -> 100
Highly volatile even at room temperature.
35-45 100 -> 200
S o lu b ility
Slightly soluble in water, very soluble in alcohol and in light 45-55 200
petroleum , freely soluble in fatty oils, very slightly soluble in Injection port 220
glycerol. Detector 250
ID E N T IF IC A T IO N
First identification: A , C.
Detection Flam e ionisation.
Second identification A, B, D
Injection 1 pL.
A. Specific optical rotation (see Tests).
System suitability Reference solution (d).
B. M elting point (2.2.14): 175 °C to 179 °C.
— resolution: minimum 3.0 between the peaks due to bomyl
C. Infrared absorption spectrophotometry (2.2.24). acetate and to linalol.
Comparison racemic camphor CRS. Limits:
D. Dissolve 1.0 g in 30 m L of methanol R. Add 1.0 g of — bomeol: not m ore than the area of the principal peak in
hydroxylamine hydrochloride R and 1.0 g of anhydrous sodium the chromatogram obtained with reference solution (c)
acetate R. Boil under a reflux condenser for 2 h. Allow to (2.0 p er cent),
cool and add 100 m L of water R. Filter, wash die precipitate — any other impurity: n o t more than half o f the area of the
obtained with 10 inL of water R and recrystallise from 10 m L principal peak in the chromatogram obtained with
of a mixture of 4 volumes of alcohol R and 6 volumes of reference solution (a) (0.5 per cent),
water R. T h e crystals, dried in vacuo, melt (2.2.14) at 118 °C — total of other impurities: not m ore than 4 times the area of
to 121 °C. the principal peak in the chromatogram obtained with
reference solution (a) (4.0 per cent),
TESTS
— disregard limit: 0.1 times the area of the principal peak in
Cany out the weighings and dissolution rapidly.
the chromatogram obtained with reference solution (b)
S o lu tio n S (0.05 p er cent).
Dissolve 2.50 g in 10 m L o f alcohol R and dilute to 25.0 m L
H a lo g en s
with the same solvent.
Maximum 100 ppm.
A p p e a ra n c e o f so lu tio n Dissolve 1.0 g in 10 m L of 2-propanol R in a distillation flask.
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
A dd 1.5 m L o f dilute sodium hydroxide solution R and 50 mg
A c id ity o r alk alin ity of nickel-alwnmium alloy R. H eat on a water-bath until the 2-
T o 10 m L of solution S add 0.1 m L o f phenolphthalein propanol R has evaporated. Allow to cool and add 5 m L of
solution R l. T h e solution is colourless. N o t more than water R. Mix and filter through a wet filter previously washed
0.2 m L o f 0.1 M sodium hydroxide is required to change the with water R until free from chlorides. Dilute the filtrate to
colour o f the indicator. 10.0 m L with water R. T o 5.0 m L of the solution, add nitric
S p ecific o p tic a l ro ta tio n (2.2.7) acid R dropwise until the precipitate which forms is
+ 41.0 to + 44.0, determined on solution S. redissolved and dilute to 15 m L with water R. T h e solution
complies with the limit test for chlorides (2.4.4).
1-394 Camphor 2016
H
R e sid u e o n e v a p o ra tio n (2.8.9) OH
M axim um 0.05 per cent. fT ^ t-c h 3
and enantiomer
Evaporate 2.0 g on a w ater-bath and dry in an oven at ch 3
100-105 °C for 1 h. T h e residue weighs a maximum o f i Cl 3
ch
1 mg.
W a te r
G. exo-2J3J3-trim ethylbicyclo[2.2.l]heptan-2-ol
Dissolve 1 g in 10 m L o f light petroleum R. T h e solution is
(camphene hydrate),
clear (2.2.1).
IM P U R IT IE S
ch 3
ch 3 f T ^ i - OH
and enantiomer
ch 3
H
ch 3
and enantiomer
h3c
H3C
H H . endo-2j3j3-trimethylbicyclo[2.2. l]heptan-2-ol
(methylcamphenilol),
A. 2,6,6-trim ethylbicyclo[3.1.1]hept-2-ene (a-pinene),
wrlj
OH
H
! Cl 3
ch and enantiomer
ch 3 S i p 1“
Ct, and enantiomer
CH3
★.* * * ★
Racemic Camphor ★ ★
*+ +*
ch 3
(Ph. Eur. monograph 0655)
ch 3
CH,
CH,
TESTS
Water
Dissolve 1 g in 10 m L of light petroleum R. The solution is
Carry out the weighings rapidly.
clear (2.2.1).
S o lu tio n S
Residue on evaporation
Dissolve 2.50 g in 10 m L of ethanol (96 per cent) R and
M aximum 0.05 per cent.
dilute to 25.0 m L with the same solvent.
Evaporate 2.0 g on a water-bath and diy at 100-105 °C for
A p p e a ra n c e o f so lu tio n
1 h. T he residue weighs not more than 1 mg.
Solution S is d ear (2.2.1) and colourless (2.2.2, Method II).
__________________________________________________ PhEur
A cid ity o r a lk alin ity
Dissolve 1.0 g in 10 m l. o f ethanol (96 per cent) R and add
0.1 m l, of phenolphthalein solution R l. T he solution is
colourless. N o t more than 0.2 m L o f 0.1 M sodium hydroxide
is required to change the colour o f the indicator. Candesartan Cilexetil *****
O p tic a l ro ta tio n (2.2. 7)
(Ph Eur. monograph 2573) *
—0.15° to + 0.15°, determined on solution S.
R elated su b sta n c e s
Gas chromatography (2.2.28).
Test solution Dissolve 50 m g o f the substance to be examined
in hexane R and dilute to 50.0 m L with the same solvent.
Reference solution (a) Dissolve 50 mg o f the substance to be
examined and 50 m g o f bomyl acetate R in hexane R and
dilute to 50.0 m L with the same solvent.
Reference solution (b) Dilute 1.0 m L o f the test solution to
200.0 m L with hexane R.
Column:
— sizer. I = 2 m , 0 = 2 mm;
— stationary phase: diatomaceous earth for gas C33H34N60 6 611 145040-37-5
chromatography R impregnated with 10 per cent mtm of
macrogol 20 000 R. A c tio n a n d u se
Carrier gas nitrogen for chromatography R. Angiotensis II (ATj) receptor antagonist
Flow rate 30 mlVmin. PhBr__________________________________________________________
Temperature.
— column: 130 °C; D E F IN IT IO N
— injection port and detector. 200 °C. (1 RS)-1- [ [(Cyclohexyloxy)carbonyI] oxy] ethyl 2-ethoxy-1-
[ [2'-(lH -tetrazol-5-yl) biphenyl-4-yI] methyl] -1H-
Detection Flam e ionisation.
benzimidazole-7-carboxylate.
Injection 1 |iL.
C o n te n t
Run time 3 times the retention time o f camphor.
99.0 per cent to 101.0 per cent (anhydrous substance).
System suitability:
— resolution: minimum 1.5 between the peaks due to CHARACTERS
cam phor and bom yl acetate in the chromatogram A p p e a ra n c e
obtained with reference solution (a); W hite or almost white powder.
1-396 Candesartan Cilexetil 2016
and enantiomer
and enantiomer
ch3
I. methyl 2-ethoxy-l-[[2/-(l//-tetrazol-5-yl)biphenyl-4-
yI]methyl]-lJi-benzimidazole-7-carboxylate.
PhEur
E. (1 RS ) - 1- [ [(cydohexyloxy)carbonyl]oxy]ethyl 2-ethoxy-l-
[ [2'-(1 -ethyl-1Ji-tetrazol-5-yI)biphenyl-4-yl] methyl] -IH -
benzimidazole-7-carboxylate,
1-398 Capecitabine 2016
Column:
Capecitabine ****** — size. I = 0.25 m , 0 = 4.6 mm;
(Ph. Eur. monograph 2762) * — stationary phase: end-capped octadecylsUyl silica gelfor
chromatography R (5 jxm);
o — temperature. 40 °C.
Mobile phase.
— mobile phase A: acetomtrUe R, methanol R, 0.1 per cent V/V
solution o f glacial acetic add R (5:35:60 VIVIV)’,
— mobile phase B: acetonitrUe R, 0.1 per cent V/V solution of
glacial acetic add R, methanol R (5:15:80 VIVIV);
2 0 -3 0 49 51
A c tio n a n d u se
Pyrimidine analogue; cytotoxic; treatm ent of colorectal
cancer. Flow rate 1.0 mL/min.
P h E u r ____________________________________________________________________________________________
Detection Spectrophotom eter at 250 nm.
Injection 10 pL of the test solution and reference solutions (b)
D E F IN IT IO N
and (c).
Pentyl [l-(5-deoxy-|3-D-ribofuranosyl)-5-fluoro-2-oxo-l,2-
dihydropyrimidin-4-yl] carbamate.
Identification of impurities U se the chromatogram obtained
with reference solution (c) to identify the peaks due to
C o n te n t impurities A, B and D.
98.0 per cent to 102.0 per cent (anhydrous substance).
Relative retention W ith reference to capecitabine (retention
CHARACTERS time = about 17 min): impurity A = about 0.18;
A p p e a ra n c e impurity B = about 0.19; impurities D and E = about 0.95.
W hite or alm ost white powder. System suitability: reference solution (c):
S o lu b ility — resolution: m inim um 1.5 between the peaks due to
Sparingly soluble in water, freely soluble in anhydrous impurities A and B; m inim um 2.0 between the peaks due
ethanol, practically insoluble in heptane. to impurity D and capecitabine.
ID E N T IF IC A T IO N
Calculation of percentage contents:
— for each impurity, use the concentration o f capecitabine
A. Specific optical rotation (see Tests).
in reference solution (b);
B. Infrared absorption spectrophotometry (2.2.24). — correction factor, multiply the peak area o f impurity B by
Comparison capecitabine CRS. 1.3.
TESTS Limits:
S p ecific o p tic a l ro ta tio n (2.2.7) — impurities A, B: for each impurity, maximum 0.3 per cent;
+ 96.0 to + 100.0 (anhydrous substance). — sum of impurities D and E: m aximum 0.2 p er cent;
Dissolve 0.250 g in methanol R and dilute to 25.0 mT. with — unspecified impurities: for each impurity, maximum
the same solvent. 0.05 per cent;
— total: maximum 0.5 p er cent;
R e la te d su b s ta n c e s — reporting threshold: 0.03 per cent.
L iquid chrom atography (2.2.29). Prepare the solutions
immediately before use or store them at 2-8 °C. Water (2.5.32)
Maximum 0.3 per cent.
Solvent mixture acetomtrUe R, methanol R, water R
Inject 1.0 m L o f a 0.200 g/m L solution of the substance to
(5:35:60 V/V/V).
be examined in methanol R.
Test solution Dissolve 60.0 m g o f the substance to be
examined in 80 m L o f the solvent mixture, sonicate until Sulfated ash (2.4.14)
dissolution is complete and dilute to 100.0 m L with the Maximum 0.1 per cent, determ ined on 1.0 g in a platinum
solvent mixture. crucible.
Reference solution (a) Dissolve 60.0 m g o f capecitabine CRS in ASSAY
80 m L of the solvent mixture, sonicate until dissolution is Liquid chromatography (2.2.29) as described in the test for
com plete and dilute to 100.0 m L with the solvent mixture. related substances with the following modification.
Reference solution (b) Dilute 1.0 m L o f the test solution to Injection T est solution and reference solution (a).
100.0 m L w ith the solvent mixture. D ilute 1.0 m L o f this Calculate the percentage content o f C 15H 22F N 3O 6 taking
solution to 20.0 m L with the solvent mixture. into account the assigned content o f capecitabine CRS.
Reference solution (c) Dissolve 3 mg of capecitabine IMPURITIES
impurity A CRS, 3 m g o f capecitabine impurity B CRS and
Spedfied impurities A, B, D , E
5 m g of capecitabine impurity D CRS in 80 mT, of the solvent
m ixture, sonicate until dissolution is complete and dilute to Other detectable impurities (the following substances would, if
100.0 m L w ith the solvent mixture. D ilute 1 m L o f the present at a sufficient level, be detected by one or other of
solution to 50 m L w ith the test solution. the tests in the m onograph. T hey are limited by the general
2016 Caprylocaproyl Macrogolglycerides 1-399
NH2
OH OH
N
O ^N E. 3-methyIbutyl [l-(5-deoxy-P-D-ribofuranosyl)-5-fluoro-2-
CH3 n oxo-1j2-dihydropyrimidin-4-yl] carbamate,
OH OH
A. 4-am ino-1-(5-deoxy-P-D-ribofuranosyl)-5-fluoropyrimidin-
2 (1 //)-one (5'-deoxy-5-fluorocytidine)3
3 0 "
O ' N
G. pentyl [l-(2,3-di-0-acetyi-5-deoxy-P-D-ribofiiranosyl)-5-
C . 1-(2j3-di-0-acetyi-5-deoxy-P-D-ribofuranosyI)-4-amino-5- fluoro-2-oxo-1,2-dihydropyrimidin-4-yl] carbamate.
fluoropyrim idin-2(l//)-one3 PhEur
Caprylocaproyl ★ ★
★ ★
Macrogolglycerides *****
(Ph. Eur. monograph 1184)
P h E tr ___________________
P e ro x id e v a lu e (2.5.5, Method A)
Maximum 6.0, determ ined on 2.0 g.
S a p o n ific a tio n v alu e (2.5.6)
Use 2.0 g.
2016 Captopril 1-401
D ilute 4 m L o f the solution to 50.0 m L with the solvent — unspecified impurities: for each im purity, n o t m ore than the
mixture. area o f die principal peak in the chrom atogram obtained
Reference solution (c) In order to prepare impurity A in situ, w ith reference solution (d) (0.10 per cent);
introduce 1.0 m L of the test solution into a volumetric flask — total: maximum 1.2 p er cent;
and add 230 jiL of 0.05 M iodine. If the solution is not — disregard Omit: 0.5 times the area o f the principal peak in
colourless, add 0.1 M sodium tkiosulfate dropwise until it the chrom atogram obtained with reference solution (d)
becomes colourless, and dilute to 50.0 m L with the solvent (0.05 per cent).
mixture. D ilute 5.0 m L o f this solution to 20.0 m L with the H e a v y m e ta ls (.2.4.8)
solvent mixture. M axim um 20 ppm .
Reference solution (d) Dilute 1.0 m L o f the test solution to Solvent water R.
100.0 m L with the solvent mixture. Dilute 1.0 m L o f this 0.50 g complies with test H . Prepare the reference solution
solution to 10.0 m L w ith the solvent mixture. using 1 m L o f lead standard solution (10 ppm Pb) R.
Column:
L o ss o n d ry in g (2.2.32)
— sizer. I = 0.3 m, 0 = 3.9 mm;
M axim um 1.0 per cent, determ ined on 1.000 g by drying
— stationary phase, end-capped octadecylsUyl silica gelfor
u n d er high vacuum at 60 °C for 3 h.
chromatography R (10 pm);
— temperature: 50 °C. S u lfa te d a s h (2.4.14)
Mobile phase: M axim um 0.2 per cent, determ ined on 1.0 g.
— mobile phase A: phosphoric add R, water R (0.08:100 VIV); A SSA Y
— mobile phase B: phosphoric add R, acetonitrile R l, water R Dissolve 0.150 g in 30 m L o f water R. T itrate with 0.05 M
(0.08:50:50 V/V/V); iodine, determining the end-point potentiom etrically (2.2.20).
U se a combined platinum electrode.
Time Mobile phase A Mobile phase B 1 m L o f 0.05 M iodine is equivalent to 21.73 m g of
(min) (per cent V7V) (percent V7V) c 9h 15n o 3s .
0 -5 90 10 IM P U R IT IE S
5 -2 0 90-» 50 10-» 50 Specified impurities A , B, C, D, E, F, J.
2 0 -4 5 50 50 Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the m onograph. T hey are limited by the general
Flow rate 1.5 miVmin acceptance criterion for other/unspecified impurities and/or
Detection Spectrophotom eter at 210 run. by the general m onograph Substances for pharmaceutical use
Irgecdon 25 |iL. (2034). It is therefore n o t necessary to identify these
impurities for demonstration o f compliance. See also 5.10.
Identification of impurities Use the chromatogram obtained Control of impurities in substances for pharmaceutical use): G, H,
with reference solution (a) to identify the peaks due to
1, L, M, N , O.
impurities B, C , D and J; use the chromatogram obtained
with reference solution (b) to identify the peak due to
im purity E; use the chrom atogram obtained with reference
solution (c) to identify the peak due to impurity A.
Relative retention W ith reference to captopril (retention
time = about 15 min): impurity C = about 0.6;
im purity D = about 0.8; impurity E = about 0.9; A. 1 ,1 '-[disulfanediylbis [(2S)-2-m ethyl-1-oxopropane-3,1-
im purity B = about 1.17; im purity J = about 1.22; diyl]]bis[(25)-pyrrolidine-2-carboxylic] acid (captopril
im purity A = about 1.7. disulfide),
System suitability:
— resolution: m inim um 1.5 betw een the peaks due to
, c o 2h
impurities B and J in the chromatogram obtained with
reference solution (a);
— resolution: m inim um 2.0 between the peaks due to
impurity E and captopril in the chromatogram obtained
w ith reference solution (b). B. (2S)-1 - [(2S)-3-bromo-2-methylpropanoyl]pyrrolidine-2-
Limits: carboxylic a d d ,
— impurity A: not m ore than 10 times the area of the
principal peak in the chrom atogram obtained with
no and enantiomer
reference solution (d) (1.0 per cent); H CH3
— impurity J: not m ore than 2.5 times the area of the
corresponding peak in the chrom atogram obtained with
C. (2itS)-2-methyl-3-sulfanylpropanoic a d d ,
reference solution (a) (0.2 p er cent);
— impurities B, C, D: for each impurity, not more than
1.5 times the area o f the corresponding peak in the and enantiomer
chrom atogram obtained w ith reference solution (a) H CH3
(0.15 per cent);
— impurity E: not m ore than 1.5 times the area of the D . (2i?5)-3-bromo-2-methylpropanoic a d d ,
principal peak in the chrom atogram obtained with
reference solution (d) (0.15 per cent);
2016 Carbachol 1-403
h o 2c . f h3c ch3
V ..CO2H
n' yç 's '^ s ' y: n'
~ & r H CH 3 H CH3
F. (25)-l-[(2Æ)-2-methyl-3-sulfanylpropanoyl]pyrrolidine-2- ★
*** ★
carboxylic acid (ept-captopril),
Carbachol ★ ★
(Ph. Eitr. monograph 1971) *****
o
G . (2i?5)-3-(acetylsulfanyl)-2-methylpropanoic acid,
C<>H15C 1N20 2 182.7 51-83-2
A ctio n a n d u se
Cholinoceptor agonist.
PhEtr.
A SSA Y
Comparison carbamazepine CRS.
Dissolve 0.150 g in a mixture of 10 m L of anhydrous acetic Preparation Examine the substances as discs without prior
acid R and 40 m L o f acetic anhydride R. Titrate with 0.1 M treatm ent.
perchloric acid. Determ ine the end-point potentiometrically TESTS
(2.2.20). Acidity or alkalinity
1 m L o f 0.1 M perchloric acid is equivalent to 18.27 m g of T o 1.0 g add 20 m L of carbon dioxide-free water R, shake for
C o H is C lN ^ . 15 min and filter. T o 10 m L o f the filtrate add 0.05 m L of
STORAGE
phenolphthalein solution R1 and 0.5 m L of 0.01 M sodium
hydroxide; the solution is red. Add 1.0 m L o f 0.01 M
In an airtight container, protected from light
hydrochloric acid; the solution is colourless. Add 0.15 m L of
IM P U R IT IE S methyl red solution R; the solution is red.
R e la te d su b sta n c e s
“ h 3c ch3
n: or Liquid chromatography (2.2.29).
HO CH3 Test solution (a) Dissolve 60.0 mg of the substance to be
examined in methanol R2 and dilute to 20.0 m L with the
A. 2-hydroxy-NjNjN-trimethylethanaminium chloride same solvent Sonicate. Dilute 10.0 m L o f this solution to
(choline chloride). 20.0 m L with water R.
--------------------------------------------------------------------- --------------------------PhEur Test solution (b) Dilute 10.0 m L o f test solution (a) to
50.0 m L with a mixture of equal volumes o f methanol R2 and
water R.
Reference solution (a) Dissolve 7.5 m g o f carbamazepine CRS,
7.5 m g o f carbamazepine impurity A CRS and 7.5 m g of
iminodibenzyl R (impurity E) in methanol R2 and dilute to
100.0 m L with the same solvent. Dilute 1.0 m L o f this
solution to 50.0 m L with a mixture o f equal volumes of
methanol R2 and water R.
2016 Carbamazepine 1-405
L o ss o n d ry in g (2.2.32)
M axim um 0.5 p er cent, determ ined on 1.000 g by drying in
an oven at 105 °C for 2 h.
Sulfated ash (2.4.14)
M axim um 0.1 per cent, determ ined on 1.0 g.
ASSAY
L iquid chromatography (2.2.29) as described in the test for
E. 10,11 -dihydro-5/i-dibenzo [bj\azepine (iminodibenzyl),
related substances with die following modifications.
Injection T est solution (b) and reference solution (b).
System suitability:
— repeatability: reference solution (b). i a
Calculate the percentage content o f C 15H 12N 20 from the
declared content o f carbamazepine CRS.
STORAGE
In an airtight container. F. 5H-dibenzo [bj\ azepine-5-carbonyl chloride
(5-chlorocarbonyliminostiIbene),
1-406 Carbaryl 2016
Loss on drying
W hen dried to constant weight over phosphorus pentoxide at a
pressure not exceeding 0.7 kPa, loses not m ore than 0.5% of
its weight. Use 1 g.
ASSA Y
Carry out the m ethod for liquid chromatography,
G. 10-brom o-5if-dibenzo [bj\ azepine- 5-carboxamide Appendix IE D , using the following solutions.
( 10-bromocarbam azepine). (1) 0.005% w/v o f the substance being examined in methanol.
PhEtr (2) 0.005% w/v of carbaryl BPCRS in methanol.
(3) 0.005% w/v of die substance being examined and
0.005% w/v of 1-naphthol in the mobile phase.
CHROMATOGRAPHIC CONDITIONS
P h E u r.
D E F IN IT IO N
(25)-3-(3,4-Dihydroxyphenyl)-2-hydrazino-2-
methylpropanoic a d d monohydrate.
C o n te n t
98.5 per cent to 101.0 per cent (dried substance).
A. 2-(acetyloxy)benzoic anhydride,
CHARACTERS
A p p e a ra n c e
^ V C02H o
UL 0
„ a ,c h 3
o
W hite o r yellowish-white powder.
S o lu b ility
Slightly soluble in water, very slightly soluble in ethanol
(96 per cent), practically insoluble in methylene chloride.
It dissolves in dilute solutions o f mineral adds.
ID E N T IF IC A T IO N
B. 2-[[2-(acetyloxy)benzoyI]oxy]benzoic a d d First identification A , C.
(acetylsalicylsalicylic ad d ),
Second identification A, B, D, E.
C02H A. Specific optical rotation (see Tests).
B. Ultraviolet and visible absorption spectrophotom etry
a (2.2.25).
Test solution Dissolve 50 m g in an 8.5 g/L solution o f
C . 2-hydroxybenzenecarboxylic a d d (salicylic ad d ), hydrochloric acid R in methanol R and dilute to 100.0 m L with
the same solution. Dilute 10.0 m L o f this solution to
100.0 m l, with an 8.5 g/L solution o f hydrochloric add R in
methanol R.
a " O OH Spectral range 230-350 nm .
Absorption maximum At 283 nm .
Specific absorbance at the absorption maximum 135 to 150
(dried substance).
C. Infrared absorption spectrophotometry (2.2.24).
D . 2-[(2-hydroxybenzoyl)oxy]benzoic a d d (salicylsalicylic Comparison carbidopa CRS.
ad d ).
D . Shake vigorously about 5 mg with 10 m L o f water R for
PhEur 1 mfn and add 0.3 m L o f ferric chloride solution R2.
A n intense green colour is produced, which quickly changes
to reddish-brown.
E. Suspend about 20 mg in 5 m L o f water R and add 5 m L
o f cupri-tartaric solution R. O n heating, the colour o f the
solution changes to dark brown and a red predpitate is
formed.
TESTS
A p p e a ra n c e o f so lu tio n
T he solution is d e a r (2.2.1) and n o t more intensdy coloured
than reference solution BY6 or B6 (2.2.2, Method II).
Dissolve 0.25 g in 25 m L o f 1 M hydrochloric acid.
S pecific o p tic a l ro ta tio n (2.2.7)
-2 6 .5 to -2 2 .5 (dried substance).
2016 Carbidopa 1-409
o
H eav y m e ta ls (2.4.8)
M axim um 20 ppm.
1.0 g complies with test C. Prepare the reference solution
using 2 m L o f lead standard solution (10 ppm Pb) R.
L oss o n d ry in g (2.2.32)
6.9 p er cent to 7.9 p er cent, determined on 1.000 g by
drying in an oven at 105 °C.
S u lfa te d a s h (2.4.14) D . methyl (2<S)-2-(2-cydohexyiidenehydrazino)-3-(3,4-
M axim um 0.1 per cent, determ ined on 1.0 g. dihydroxyphenyl)-2-m ethylpropanoate,
A SSA Y
Dissolve 0.150 g with gentle heating in 75 m L of anhydrous o
acetic add R. T itrate with 0.1 M perchloric add, determining
the end-point potentiometrically (2.2.20).
1 m L o f 0.1 M perchloric acid is equivalent to 22.62 mg
of C 10H 14N 2O 4.
STORAGE
Protected from light. E. methyl (25)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2-
m ethylpropanoate,
IM P U R IT IE S
Spedfied impurities A, D , E, F, H , I, J
o
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
oh
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10. F. ethyl (25)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2-
Control of impurities in substances for pharmaceutical use): B, C, methylpropanoate,
G.
^ / \ X 0 2H
HO
JL J h2n ch3
OH
OH
G. l-(3,4-dihydroxyphenyl)propan-2-one,
A. (26)-2-amino-3-(3,4-ciihydroxyphenyl)-2-methylpropanoic
acid (L-methyldopa),
o
JL Jl HN ’CH3
h3c o nh2
OH
JL Jl h2n 'c h 3
. ho H . (25) - 2 -hydrazina-3-(3-hydroxy-4-methoxyphenyl)-2-
OH methylpropanoic add,
B. methyl (26)-2-amino-3-(3,4-dihydroxyphenyl)-2-
methylpropanoate,
Jl J HN CH3
nh2
OH
JL J HN CH3
HO Y nh2 I. (25)-3-(3-bromo-4,5-dihydroxyphenyl)-2-hydrazino-2-
och3 methylpropanoic ad d ,
C. (25)-2-hydrazino-3-(4-hydroxy-3-methoxyphenyl)-2- Br
methylpropanoic acid (3-O-methylcarbidopa),
I J HN CH3
ho' Y
OH
J. (25)-3-(2-bromo-4,5-dihydroxyphenyl)-2-hydrazino-2-
methylpropanoic add.
PhEur
2016 Carbimazole 1-411
r~ \ N in h y d rin -p o sitiv e su b s ta n c e s
h 3c ' Y Examine by thin-layer chromatography (2.2.27), using a
SH suitable silica gel as the coating substance.
Test solution (a) Dissolve 0.10 g o f the substance to be
A. 1-m ethyl-lH-imidazole-2-thiol (thiamazole). examined in dilute ammonia R2 and dilute to 10 mT. with the
PhEur same solvent.
Test solution (b) Dilute 1 m L o f test solution (a) to 50 mT.
with water R.
Reference solution (a) Dissolve 10 m g of carbocisteine CRS in
***** dilute ammonia R2 and dilute to 50 m L with the same
Carbocisteine * * solvent.
* * * * * Reference solution (b) D ilute 5 m L o f test solution (b) to
(Ph. Ever, monograph 0885)
20 m L with water R.
H NH, Reference solution (c) Dissolve 10 m g of carbocisteine CRS and
HOjC. 10 mg of arginine hydrochloride CRS in 5 m L o f dilute
COjH
ammonia R2 and dilute to 25 m L with water R.
Apply separately to the plate 5 (jL of each solution. Allow the
C5H 9N 04S 179.2 638-23-3
plate to dry in air. Develop over a path of 15 cm using a
A ctio n a n d u se mixture of 20 volumes o f glacial acetic add R, 20 volumes of
Mucolytic. water R and 60 volumes o f butanol R. Dry the plate in a
current o f warm air. Spray with ninhydrin solution R and heat
P h E u r ____________________ at 100 °C to 105 °C for 15 min. Any spot in the
D E F IN IT IO N chromatogram obtained with test solution (a), apart from the
principal spot, is not m ore intense than the spot in the
Carbocisteine contains not less than 98.5 per cent and not
chromatogram obtained with reference solution (b)
m ore than the equivalent o f 101.0 per cent o f (2i?)-2-amino-
(0.5 per cent). T he test is not valid unless the chromatogram
3-[(carboxymethyl)sulfanyl]propanoic acid, calculated with
obtained with reference solution (c) shows two clearly
reference to the dried substance.
separated principal spots.
CHARACTERS
C h lo rid e s (2.4.4)
A white or alm ost white, crystalline powder, practically
Dissolve 33 mg in 5 m L o f dilute nitric acid R and dilute to
insoluble in w ater and in alcohol. It dissolves in dilute
15 m L with water R. T he solution, without further addition
mineral a d d s and in dilute solutions o f alkali hydroxides.
o f nitric acid, complies with the limit test for chlorides
ID E N T IF IC A T IO N (0.15 per cent).
First identification A, B. S u lfa tes (2.4.13)
Second identification A , C, D. Dissolve 0.5 g in 5 m L o f dilute hydrochloric add R and dilute
A. Specific optical rotation (see Tests). to 15 m L with distilled water R. T h e solution complies with
B. Examine by infrared absorption spectrophotometry the limit test for sulfates (300 ppm ).
(2.2.24), comparing with the spectrum obtained with H eav y m e ta ls (2. 4.8)
carbocisteine CRS. Examine the substances prepared as discs. 2.0 g complies with test D for heavy metals (10 ppm).
C. Examine the chromatograms obtained in the test for Prepare the reference solution using 2 m L o f lead standard
ninhydrin-positive substances. T he principal spot in the solution (10 ppm Pb) R
chrom atogram obtained with test solution (b) is similar in L oss o n d ry in g (2.2.32)
position, colour and size to the principal spot in the N o t more than 0.5 per cent, determined on 1.000 g by
chrom atogram obtained with reference solution (a). drying in an oven at 105 °C for 2 h.
D . Dissolve 0.1 g in 4.5 m L o f dilute sodium hydroxide S u lfa te d a s h (2.4.14)
solution R H eat on a water-bath for 10 min. Cool and add N o t more than 0.3 per cent, determined on 1.0 g.
1 m L of a 25 g/L solution o f sodium nitroprusside R A dark
ASSAY
red colour is produced, which changes to brown and then to
Dissolve 0.150 g in 10 m L o f anhydrous formic add R with
yellow within a few minutes.
slight heating and shake until dissolution is complete.
TESTS Add 50 m L o f anhydrous acetic add R. T itrate with 0.1 M
S o lu tio n S perchloric add, determining the end-point potentiometrically
Disperse 5.00 g in 20 m L o f water R and add dropwise with (2.2.20).
shaking 2.5 m L of strong sodium hydroxide solution R. Adjust 1 m L o f 0.1 M perchloric add is equivalent to 17.92 m g of
to p H 6.3 with 1 M sodium hydroxide and dilute to 50.0 m L C sH ^S .
w ith water R.
STO R A G E
A p p e a ra n c e o f so lu tio n
Store protected from light.
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
_________________________________________________________________ PhEur
p H (2.2.3)
Shake 0.2 g with 20 m L o f carbon dioxide-free water R.
T h e pH of the suspension is 2.8 to 3.0.
S pecific o p tic a l ro ta tio n (2.2.7)
—32.5 to -3 5 .5 , determ ined on solution S and calculated
w ith reference to the dried substance.
2016 Carbomers 1-413
DEFINITION 9 -2 0 0 100
High-molecular-mass polymers of acrylic acid cross-linked
w ith alkenyl ethers o f sugars or polyalcohols.
Flow rate 1 mL/min.
Content Detection Spectrophotometer at 205 nm.
56.0 per cent to 68.0 per cent o f carboxylic acid (-C 0 2H)
Injection 20 jiL.
groups (dried substance).
Retention lime Acrylic acid = about 6.0 min.
CHARACTERS
Limit:
Appearance
— acrylic acid: not more than the area of the corresponding
W hite or almost white, fluffy, hygroscopic powder.
peak in the chromatogram obtained with the reference
Solubility solution (0.25 per cent).
Swells in water and in other polar solvents after dispersion
B en z e n e
and neutralisation w ith sodium hydroxide solution.
G as chromatography (2.4.24, System A).
ID E N T IF IC A T IO N Solution A Dissolve 0.100 g of benzene R in dimethyl
First identification A sulfoxide R and dilute to 100.0 m L with the same solvent.
Second identification B, C} D D ilute 1.0 m L of the solution to 100.0 m L with water R.
A. Infrared absorption spectrophotometry (2.2.24). Dilute 1.0 m L o f this solution to 100.0 m L with water R.
Main bands A t 1710 ± 5 cm-1, 1454 ± 5 cm -1, Test solution Weigh 50.0 m g of the substance to be examined
1414 ± 5 cm -1 , 1245 ± 5 cm-1, 1172 ± 5 cm-1, into an injection vial and add 5.0 m L of water R and 1.0 m L
1115 ± 5 cm-1 and 801 ± 5 cm-1 , with die strongest band o f dimethyl sulfoxide R.
at 1710 ± 5 cm -1 . Reference solution Weigh 50.0 mg o f the substance to be
B. Adjust a 10 g/L dispersion to about p H 7.5 with examined into an injection vial and add 4.0 m L o f water R,
1 M sodium hydroxide. A highly viscous gel is formed, 1.0 m L o f dimethyl sulfoxide R and 1.0 m L o f solution A.
c . Add 2 m L o f a 100 g/L solution of cakiian chloride R, Close the vials with a tight rubber membrane stopper coated with
with continuous stirring, to 10 m L o f die gel from polytetrafiuoroethylene and secure with an afambuum crimped
identification test B. A white precipitate is immediately cap. Shake to obtain a homogeneous dispersion.
produced. Static head-space conditions that may be used:
D . Add 0.5 m L o f thymol blue solution R to 10 m L o f a — equilibration temperature: 80 ° Q
10 g/L dispersion. A n orange colour is produced. — equilibration timer. 60 min;
Add 0.5 m L o f cresol red solution R to 10 m L of a 10 g/L — transfer-line temperature: 90 °C.
dispersion. A yellow colour is produced. Injection 1 m L o f the gaseous phase o f the test solution and
1 m L of the gaseous phase of the reference solution; repeat
TESTS
these injections twice more.
Free acrylic acid
Liquid chromatography (2.2.29).
System suitability:
— repeatability: maximum relative standard deviation of the
Test solution M ix 0.125 g of die substance to be examined differences in area between the analyte peaks obtained
with a 25 g/L solution of aluminium potassium sulfate R and
from the 3 replicate pair injections of the reference
dilute to 25.0 m L w ith the same solution. H eat the
solution and the test solution is 15 per c e n t
suspension at 50 ° c for 20 min with shaking, then shake die
Limit:
suspension at room temperature for 60 min. Centrifuge and
— benzene: the m ean area o f the peak due to benzene in the
use the clear supernatant solution as the test solution.
chromatograms obtained with the test solution is not
Reference solution Dissolve 62.5 mg of acrylic acid R in a greater than 0.5 times the m ean area of the peak due to
25 g/L solution of aluminium potassium sulfate R and diỉutè to benzene in the chromatograms obtained with the
100.0 m L with the same solution. Dilute 1.0 m L of this reference solution (2 ppm).
solution to 50.0 m L with a 25 g/L solution o f aluminium
H eav y m e ta ls (2.4.8)
potassium sulfate R.
M aximum 20 ppm .
Column:
— size. I = 0.12 m , 0 = 4.6 mm; 1.0 g complies w ith test C. Prepare the reference solution
— stationary phase’, octadecyisHyl silica gelfor chromatography R using 2 m L of lead standard solution (10 ppm Pb) R.
(5 pm). L o ss o n d ry in g (2.2.32)
Mobile phase: M axim um 3.0 per cent, determined on 1.000 g by drying
— mobile phase A'. 1.361 g/L solution OĨpotassium dihydrogen in vacuo at 80 °C for 60 m in.
phosphate R, adjusted to p H 2.5 using dilute phosphoric S u lia te d ash (2.4.14)
acid R ; M aximum 4.0 per cent, determined on 1.0 g.
1-414 Carbon Dioxide 2016
A SSA Y
Carbon Dioxide *****
Slowly add 50 m L o f water R to 0.120 g whilst stirring and *. ★
heating at 60 °c for 15 min. Stop heating, add 150 m L o f (Ph. Eur. monograph 0375) *
water R and continue stirring for 30 min, Add 2 g o f
C 02 44.01 124-38-9
potassium chloride R and titrate with 0.2 M sodium hydroxide,
determining the end-point potentíometrically (2.2.20). Carbon Dioxide should be kept in approved metal cylinders which
are painted grey and cany a label stating ‘Carbon Dioxide3.
1 m L of 0.2 M sodium hydroxide is equivalent to 9.0 m g of
In addition, cCarbon Dioxide ’ or the symbol ‘C02 ’ should be
carboxylic acid (-C 0 2H) groups.
stencilled in paint on the shoulder of the cylinder.
STORAGE P h E u r ___________________________________________________________________________________________
In an airtight container.
D E F IN IT IO N
F U N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S C o n te n t
This section provides information on characteristics that are M inim um 99.5 per cent V/V of C 0 2 in the gaseous phase.
recognised as being relevant control parameters for one or more
This monograph applies to carbon dioxide for medicinal use.
junctions of the substance when used as an excipient (see chapter
5.15). Thừ section ừ a rum-mandatory part of die monograph CHARACTERS
and it is not necessary to verify the characteristics to demonstrate A p p e a ra n c e
compliance. Control of these characteristics can however contribute Colourless gas.
to the quality of a medừinơỉ product by improving the consistency S o lu b ility
of the manufacturing process and the performance of the medicinal At 20 °C and at a pressure o f 101 kPa, 1 volume dissolves in
product during use. Where control methods are coed, they are about 1 volume o f water.
recognised as being suừabỉe for the purpose, but ođier methods can
P R O D U C T IO N
also be used. Wherever results for a particular characteristic are
reported, the control method, must be indicated. Examine the gaseous phase.
The following characteristics may be relevant for carbomers used as I f the testis performed on a cylinder of gas, keep the cylinder of
viscosity-increasing agents and gelling agents. the substance to be examined at room temperature for not less than
6 h before carrying out the tests. Keep the cylinder in the vertical
A p p a re n t visco sity (2.2. 10)
position with the outlet valve uppermost.
T he nominal apparent viscosity is typically between
300 m P a s and 115 000 mPa-s. F or a product with a C a rb o n m o n o x id e
nom inal apparent viscosity of 20 000 mPa-s or greater, die Gas chromatography (2.2.25).
apparent viscosity is typically 70.0 per cent to 130.0 per cent Gas to be examined T he substance to be examined.
o f die nominal value; for a product with a nominal apparent Reference gas A mixture containing 5 ppm V/V o f carbon
viscosity of less than 20 000 mPa-s, the apparent viscosity is monoxide R in nitrogen R l.
typically 50.0 per cent to 150.0 per cent of the nominal Column:
value. — material: stainless steel,
D ry the substance to be examined in vacuo at 80 °c for 1 h. — size. I — 2 m , 0 = 4 mm,
Carefully add 2.50 g of the previously dried substance to be — stationary phase, an appropriate molecular sieve for
examined to 500 m L of water R in a 1000 m L beaker while chromatography (0.5 nm).
stirring continuously at 1000 ± 50 r/min, with the stirrer Carrier gas helium for chromatography R.
shaft set at an angle o f 60° to one side o f the beaker. Add the
Flow rate 60 mL/min.
previously dried substance over a period of 45-90 s, at a
Temperature.
uniform rate, ensuring that loose agglomerates o f powder are
— column: 50 °C,
broken up, and continue stirring at 1000 ± 50 r/min for
— injection port and detector. 130 °C.
15 min. Remove die stiirer and place die beaker containing
die dispersion in a water-bath at 25 ± 1 °c for 30 m in. Detection Flame ionisation with methaniser.
Insert the stirrer to a depth necessary to ensure that air is n o t Injection Loop injector.
draw n into the dispersion and, while stirring at Adjust the injected volumes and the operating conditions so
300 ± 25 r/min, tìơate with a glass-calomel electrode system that the height o f the peak due to carbon monoxide in the
to p H 7.3-7.8 by adding a 180 g/L solution of sodium chromatogram obtained with the reference gas is at least
hydroxide R below die surface, determining die end-point 35 p er cent o f the full scale of the recorder.
potentiometrically (2.2.2Ơ). T h e total volume of the Ì80 g/L Limit.
solution o f sodium hydroxide R used is about 6.2 m ĩ- Allow — carbon monoxide, not more than the area o f the
2-3 min before the final p H determination. If the final p H corresponding peak in the chrom atogram obtained with
exceeds 7.8, discard the preparation and prepare another the reference gas (5 ppm V/V).
using a smaller am ount o f sodium hydroxide for titration.
N itro g e n m o n o x id e a n d n itro g e n d io x id e
R eturn die neutralised preparation to die water-bath at 25 °c
M axim um 2 ppm V/V in total, determ ined using a
for 1 h, then perform the viscosity determination without
delay to avoid slight viscosity changes that occur 75 min after chemiluminescence analyser (2.5.26).
neutralisation. Determ ine die viscosity using a rotating Gas to be examined T he substance to be examined.
viscometer with a spindle rotating at 20 r/min, using a Reference gas (a) Carbon dioxide R l.
spindle suitable for the expected apparent viscosity. Reference gas (b) A mixture containing 2 ppm V/V o f nitrogen
C arboxylic a c id g ro u p s monoxide R in carbon dioxide R l or in nitrogen R l.
See Assay. Calibrate the apparatus and set the sensitivity using reference
PhEur gases (a) and (b). M easure the content o f nitrogen monoxide
and nitrogen dioxide in the gas to be examined.
2016 Carbon Dioxide 1-415
t t
Photomultiplier Amplifier
PhEur
Test solution (b) Dilute 1 m L of test solution (a) to 10 m L
with methylene chloride R.
Reference solution (a) Dissolve 5.0 mg o f meprobamate CRS in
methylene chloride R and dilute to 50 m L with the same
solvent.
Reference solution (b) D ilute 1 m L of test solution (b) to
50 m L with methylene chloride R.
1-420 Carmellose 2016
o
Reference solution (c) Dilute 5 m L o f reference solution (b) to
10 m l. with methylene chloride R. o.A.o
Reference solution (d) Dissolve 20 mg of carisoprodol CRS in
methylene chloride R and dilute to 10 m L with the same
h 3c
solvent.
ch3
Reference solution (e) Dissolve 10 mg o f carisoprodol
impurity A CRS in 5 m L of reference solution (d) and dilute B. 5-methyl-5-propyl-l,3-dioxan-2-one,
to 50 m L with methylene chloride R.
Plate TLC silica gel plate R.
Mobile phase acetone R, methylene chloride R (20:80 V/V). h 3c X v
Application 5 |iL. ch3
and the washings and dilute to 100 m L with distilled water R. A p p a re n t v iscosity
T o 25 m L add 1 m L of dilute hydrochloric add R and dilute While stirring, introduce a quantity o f the substance to be
to 50 m L with distilled water R. examined equivalent to 2.00 g o f the dried substance into
H eav y m e ta ls (2.4.8) 50 m L o f water R heated to 90 °C. F or a product o f low
M axim um 20 ppm. viscosity, use if necessary, the quantity required to give the
concentration indicated on the label. Allow to cool, dilute to
1.0 g complies with test D . Prepare the reference solution
100.0 m L with water R and stir until dissolution is complete.
using 2 m L o f lead standard solution (10 ppm Pb) R.
Determ ine the viscosity (2.2.10) using a rotating viscometer
L oss o n d ry in g (2.2.32) at 20 °C and a shear rate o f 10 s-1 . If it is impossible to
M aximum 10.0 per cent, determined on 1.000 g by drying in obtain a shear rate o f exactly 10 s-1 , use a shear rate slightly
an oven at 105 °C for 4 h. higher and a rate slightly lower and interpolate. T h e apparent
S u lfa te d a s h (2.4.14) viscosity is not less than 75 per cent and n o t m ore than
10.0 per cent to 20.0 per cent, determined on 1.0 g in a 140 per cent of the value stated on the label.
platinum crucible. S o d iu m glycollate
STORA GE Place a quantity o f the substance to be examined equivalent
In an airtight container. to 0.500 g o f dried substance in a beaker. A dd 5 m L of acetic
add R and 5 m L o f water R. Stir until dissolution is complete
_______________________________________________________________ PhEur
(about 30 min). Add 80 m L o f acetone R and 2 g o f sodium
chloride R. Filter through a fast filter paper impregnated with
acetone R into a volumetric flask, rinse the beaker and filter
with acetone R and dilute the filtrate to 100.0 m L with the
Carmellose Sodium ***** same solvent. Allow to stand for 24 h without shaking.
** +* Use the clear supernatant to prepare the test solution.
(Ph. Eur. monograph 0472) *
In a volumetric flask, dissolve 0.310 g of glycoBic add R,
9004-32-4 previously dried in vacuo over diphosphorus pentoxide R, in
water R and dilute to 1000.0 m L with the same solvent.
A c tio n a n d u se
Place 5.0 m L o f this solution in a volumetric flask, add 5 m L
Excipient; bulk laxative.
o f acetic add R and allow to stand for about 30 min.
P r e p a r a tio n Add 80 m L of acetone R and 2 g o f sodium chloride R and
Carmellose Sodium Eye Drops dilute to 100.0 m L with acetone R. Use this solution to
prepare the reference solution.
P h E u r ___________________________________________________________________________________________
Place 2.0 m L o f each solution in a separate 25 m L
D E F IN IT IO N volumetric flask. H eat on a water-bath to eliminate acetone.
Carmellose sodium (carboxymethylcellulose sodium) is the Cool to room tem perature and add 5.0 m L o f 2,7-
sodium salt o f a partly O-carboxymethylated cellulose. dihydroxynaphthalene solution R to each flask. Shake and add
It contains not less than 6.5 per cent and not more than 15.0 m L of 2,7-dihydroxynaphthalene solution R. Close the
10.8 per cent o f sodium (N a), calculated with reference to flasks with aluminium foil and heat on a water-bath for
the dried substance. 20 min. Cool under running water and dilute to 25.0 m L
CHARACTERS with sulfuric add R. W ithin 10 min, transfer 10.0 m L o f each
A white or alm ost white, granular powder, hygroscopic after solution to a flat-bottomed tube. Examine the solutions
drying, practically insoluble in acetone, in ethanol and in viewing vertically. T he test solution is not more intensely
toluene. It is easily dispersed in water giving colloidal coloured than the reference solution (0.4 per cent).
solutions. C h lo rid e s (2.4.4)
ID E N T IF IC A T IO N Dilute 2 mT, o f solution S to 15 m L with water R.
T h e solution complies with the limit test for chlorides -
A. T o 10 m L o f solution S (see Tests) add 1 m L of copper
(0.25 per cent).
sulfate solution R. A blue, cotton-like precipitate is formed.
B. Boil 5 m L of solution S for a few minutes. N o precipitate H eav y m e ta ls (2.4.8)
is formed. T o the residue obtained in the determination o f the sulfated
ash, add 1 m L o f hydrochloric add R and evaporate on a
C. T h e solution prepared from the sulfated ash in the test for
water-bath. Take up the residue in 20 m L o f water R. 12 m L
heavy metals gives the reactions of sodium (2.3.1).
o f the solution complies with test A for heavy metals
TESTS (20 ppm). Prepare the reference solution using lead standard
S o lu tio n S solution (1 ppm Pb) R.
Sprinkle a quantity of the substance to be examined L oss o n d ry in g (2.2.32)
equivalent to 1.0 g of the dried substance onto 90 m L of N ot more than 10.0 per cent, determined on 1.000 g by
carbon dioxide-free water R at 40 °C to 50 °C stirring drying in an oven at 105 °C.
vigorously. Continue stirring until a colloidal solution is
S u lfa te d a s h (2.4.14)
obtained, cool and dilute to 100 m L with carbon dioxide-free
20.0 per cent to 33.3 per cent, determined on 1.0 g using a
water R.
mixture o f equal volumes o f sulfuric add R and water R and
A p p e a ra n c e o f so lu tio n calculated with reference to the dried substance. T hese limits
Solution S is not more opalescent than reference correspond to a content o f 6.5 per cent to 10.8 per cent of
suspension IH (2.2.1) and not more intensely coloured than sodium (Na).
reference solution Y6 (2.2.2, Method II).
p H (2.2.3)
T he p H o f solution S is 6.0 to 8.0.
2016 Carmellose Sodium 1-423
A c tio n a n d u se
Cytotoxic alkylating agent. A. 1,3-bis (2 -chloroethyl)urea.
PhEur
P hE ur.
D E F IN IT IO N
Carm ustine contains n o t less than 98.0 per cent and n o t
m ore than the equivalent of 102.0 per cent of
1,3-bis (2-chloroethyl) -1 -nitrosourea, calculated with reference
to the anhydrous substance.
CHARACTERS
A yellowish, granular pow der, very slightly soluble in water,
very soluble in methylene chloride, freely soluble in ethanol.
It melts at about 31 °C with decomposition.
2016 Carrageenan 1-425
★ ★ A cid valu e
Camauba Wax ★ ★ 2 to 7.
★. ★
(Ph. Ever, monograph 0597) *★* T o 2.000 g (m g) in a 250 m L conical flask fitted with a
reflux condenser add 40 mT. o f xylene R and a few glass
A ctio n a n d u se beads. H eat with stirring until the substance is completely
Excipient. dissolved. Add 20 m L of alcohol R and 1 m L o f bromothymol
blue solution R3 and titrate the hot solution with
PhEur.
0.5 M alcoholic potassium hydroxide until a green colour
D E F IN IT IO N persisting for at least 10 s is obtained (n1 mL). C arry out a
Purified wax obtained from the leaves of Copemicia cerifera blank test (n2 mL). Calculate the acid value from the
M art. expression:
CHARACTERS
A p p e a ra n c e 2 8 .0 5 ( n i — n j )
Pale yellow or yellow powder, flakes or hard masses.
S o lu b ility
Practically insoluble in water, soluble on heating in ethyl S a p o n ifica tio n v alu e
acetate and in xylene, practically insoluble in alcohol. 78 to 95.
T o 2.000 g (m g) in a 250 m L conical flask fitted with a
R elativ e d e n sity
A bout 0.97. reflux condenser add 40 m L of xylene R and a few glass
beads. H eat with stirring until the substance is completely
ID E N T IF IC A T IO N dissolved. Add 20 m L o f alcohol R and 20.0 m L o f
Thin-layer chromatography (2.2.27). 0.5 M alcoholic potassium hydroxide. Boil under a reflux
Test solution Dissolve 0.10 g of the substance to be examined condenser for 3 h. Add 1 m L of phenolphthalein solution R l
with heating in 5 m L of chloroform R. Use the warm solution. and titrate the hot solution immediately with
Reference solution Dissolve 5 mg o f menthol R, 5 pL o f menthyl 0.5 M hydrochloric acid until the red colour disappears.
acetate R and 5 mg of thymol R in 10 m L of toluene R. Repeat the heating and titration until the colour no longer
reappears on heating (n3 mL). Carry out a blank test
Plate TLC silica gel plate R
(n4 m L). Calculate the saponification value from the
Mobile phase ethyl acetate R, chloroform R (2:98 VIV). expression:
Application 30 |iL of the test solution and 10 pL of the
reference solution as bands 20 m m by 3 mm. 28.05 (714 — TI3)
Development Over half of the plate. m
Drying In air. T o ta l a s h (2.4.16)
Detection Spray with a freshly prepared 200 g/L solution of M aximum 0.25 per cent, determined on 2.0 g.
phosphomolybdic acid, R in alcohol R (about 10 m L for a 20 cm
STORA GE
plate). H eat at 100-105 °C for 10-15 min.
Protected from light.
Results T he chromatogram obtained with the reference
solution shows in the lower part a dark blue zone (menthol), PhEur
ID E N T IF IC A T IO N T o ta l a s h (2.416)
A. Prepare a 20 g/L dispersion and heat in a water-bath at M axim um 40.0 p er cent.
80 °C. M ix 1 volume o f this solution and about 4 volumes of A sh in so lu b le ỉn h y d ro c h lo ric a c id (2.8.1)
water R and add 2-3 drops o f a 0.5 g/L solution o f methylene M aximum 2.0 per c e n t
blue R in ethanol (96 per cent) R. A blue precipitate is formed.
F U N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S
B. Infrared absorption spectrophotom etry (2.2.24).
This section provides information on characteristics that are ■
Preparation Prepare a 2 g/L solution o f the substance to be recognised as being relevant control parameters for one or more
examined; cast 4.5 m L o f the solution into a plastic flat- functions of the substance when used as an excipient (see chapter
bottom ed weighing b oat about 35 m m in diameter and allow 5.15). Some of the characteristics described in the Functionality-
to dry completely in an air-flow oven at 60 °C until a film, related characteristics section may also be present in the mandatory
about 10 jim thickj is obtained (about 4 h). part of the monograph since they also represent mandatory quality
C arrageenan has strong, broad absorption bands, typical o f criteria. In such cases, a cross-reference to the tests described in the
all polysaccharides, in the 1000-1100 cm -1 region. mandatory part ừ included in the Fttncttonatity-rdated
Absorption maxima are 1065 cm -1 and 1020 cm-1 for characteristics section. Control of the characteristics can contribute
gelling and non-gelling types, respectively. O ther to the quality of a medicinal product by improving the consistency
characteristic absorption bands and their intensities relative to of the manufacturing process and the performance of the medicinal
the absorbance at 1050 cm -1 are shown in Table 2138.-1. product during use. Where control methods are cited, they are
recognised as being suitable far the purpose, but other methods can
Table 2138.-1. - Characteristic absorption bands fo r also be used Wherever results for a particular characteristic are
carrageenan identification by infrared absorption reported, the control method must be indicated
spectrophotometry The following characteristics may be relevantfor carrageenan used
Absorbance relative to the as viscosity-increasing agent
Wanrennmber absorbance at 1050 cm '1 G e l fo rm a tio n
Molecular structure
(cm -1)
K 1 \ Prepare a 20 g/L dispersion and heat in a w ater-bath at
80 °c (solution A). Allow to cool; it becomes m ore viscous
0 .7 - 1.4 -
1220 - 1260 Ester sulfate
1.2
1.2 -2.0
2.0
u pon cooling and may form a gel.
T o 10 m L o f solution A, while still hot, add 4 drops o f a
3,6-Anhydro-D- 0.3 - 0.2 -
928 - 933
gaiactose 0.6 0.5
£ 0 .2 100 g/L solution of potassium chloride R, mix and allow to
cool. A ‘brittle’ gel indicates a carrageenan o f a
0.3 - 0.2 - predom inantly K-type; an ‘elastic’ gel indicates a
840 -8 5 0 Galactose-4-sulfate -
0.5 0.4
predom inantly 1-type; if the solution does n o t form a gel, the
825 - 830 Galactose-2-sulfate - -
0.2 - carrageenan is of a predom inantly X-type; a weak or pourable
0.4 gel obtained on cooling indicates a carrageenan comprising a
0.1 - m ixture o f K- and X-types, which can be confirmed by
810 - 820 GaIactose-6-sulfate - -
0.3 infrared absorption spectrophotometry.
3,6-Anhydro-D- 0.2 - V iscosity
800 - 805 <0.2 -
gaIactose-2-sulfate 0.4 (see Tests).
____________________________________________________________________________________________ PhEur
M e rc u ry (2.427) D E F IN IT IO N
M axim um 1.0 ppm. 5 - [(2RS) -3- [(1,1 -Dimethylethyl) amino] -2-hydroxypr opoxy] -
L oss o n d ry in g (2.2.32) 3,4-dihydroquinolin-2(l/i)-one hydrochloride.
M axim um 12.0 per cent, determ ined on 1.000 g by drying in C o n te n t
an oven at 105 °C. 99.0 per cent to 101.0 per cent (dried substance).
2016 Carteolol Hydrochloride 1-427
CHARACTERS Limits:
Appearance — impurity H: not more than twice the area o f the principal
W hite or almost white crystals or crystalline powder. peak in the chromatogram obtained with reference
Solubility solution (b) (0.2 per cent);
Soluble in water, sparingly soluble in methanol, slightly — unspecified impurities: for each impurity, not more Than the
soluble in ethanol 96 per cent, practically insoluble in area o f the principal peak in the chromatogram obtained
methylene chloride. with reference solution (b) (0.10 per cent);
— total: not more than half the area of the principal peak in
ID E N T IF IC A T IO N the chromatogram obtained with reference solution (a)
A. Infrared absorption spectrophotometry (2.2.24). (0.5 p er cent);
Comparison Ph. Eur. reference spectrum of carteolol hydrochloride. — disregard limit: 0.2 times the area o f the principal peak in
B. It gives reaction (a) o f chlorides (2.3.1). the chromatogram obtained with reference solution (b)
(0.02 per cent).
TESTS
L oss o n d ry in g (2.2.32)
Appearance o f solution
M axim um 0.5 per cent, determined on 1.000 g by drying in
T h e solution is clear (2.2.1) and colourless (2.2.2,
an oven a t 105 °C for 3 h.
Method II).
Dissolve 0.300 g in water R and dilute to 10 m L with the S u lfa te d a s h (2.414)
same solvent. M aximum 0.1 per cent, determined on 1.0 g.
pH (2.2.3) A SSAY
5.0 to 6.0. Dissolve 0.250 g in 60 m L of ethanol (96 per cent) R.
Dissolve 0.250 g in carbon dioxide-free water R and dilute to A dd 5.0 m L o f 0.01 M hydrochloric acid. C any out a
25 m L w ith the same solvent. potentiometric titration (2.2.20), using 0.1 M sodium
hydroxide. Read the volume added between the 2 points o f
Related substances inflexion.
Liquid chromatography (2.2.29).
1 m L of 0.1 M sodium hydroxide is equivalent to 32.88 m g of
Test solution Dissolve 20.0 mg o f the substance to be C 16H 25N 2O 3CI.
examined in the mobile phase and dilute to 10.0 m L with
the mobile phase. STORA GE
Reference solution (a) Dilute 1.0 m L of the test solution to In an airtight container.
100.0 m L with the mobile phase. IM P U R IT IE S
Reference solution (b) Dilute 1.0 m L o f reference solution (a) Specified impurities H
to 10.0 m L with the mobile phase. Other detectable impurities (the following substances would, if
Reference solution (c) Dissolve 10 m g of carteololfor system present at a sufficient level, be detected by one or other o f
suitability CRS in the mobile phase and dilute to 5 m L with the tests in the monograph. They are limited by the general
the mobile phase. acceptance criterion for other/unspecified impurities and/or
Reference solution (d) Dilute 5.0 m L o f reference solution (b) by the general monograph Substances for pharmaceutical use
to 10.0 m L with the mobile phase. (2034). I t is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Column:
— size: I = 0.25 m , 0 = 4.6 mm;
Control of impurities in substances for pharmaceutical use): A , B,
C, D, E, F, G, I.
— stationary phase: octadecylsHyl silica gel for chromatography R
(5 nm).
Mobile phase Mix 1 volume of methanol R2, 20 volumes of
acetonitrile R and 79 volumes of a 2.82 g/L solution of sodium
hexartesulfonate R.
Flow rate 1 mL/min.
Detection Spectrophotometer at 252 nm.
A. 4,6,7,8-tetrahydroquinoline-2,5(l//,3/f)-dione,
Irgecdon 20 [iL.
Identification of impurities Use the chromatogram supplied
with carteolol for system suitability CRS to identify the peak
due to impurity H .
System suitability:
— the chromatogram obtained with reference solution (c) is
similar to the chromatogram provided with carteololfor
system stdtabHay CRS; the peaks due to impurity H and B. 5-hydroxy-3,4-dihydroquinolin-2(l.H)-one,
carteolol show base-line separation;
— signal-to-noise ratio: minimum 10 for the principal peak in
the chromatogram obtained with reference solution (d); -s^ °
.-x vvJ and enantiomer
— number of theoretical plates: minimum 6000, calculated for
the principal peak in the chromatogram obtained with
reference solution (a).
C . 5-[ [(2i?S)-oxiran-2-yl]methoxy]-3,4-dihydroquinolin-
2 (1 //)-one,
1-428 Carvedilol 2016
Solubility
R Practically insoluble in water, Sparingly soluble in methylene
and enantiomer chloride, slightly soluble in ethanol (96 per cent). It is
practically insoluble in dilute adds.
R1 It shows polymorphism (5.9).
IDENTIFICATION
D . R = Cl, R ' = H: 5-[(2ÄS)-3-chloro-2-hydroxypropoxy]- Infrared absorption spectrophotometry (2.2.24).
3 j4-dihydroquinolin-2 ( 1if)-o n e, Comparison carvedüol CRS.
F. R = O C H 3, R ' = H : 5-[(2ÄS)-2-hydroxy-3- I f the spectra obtained show differences, dissolve the
methoxypropoxy] -3,4-dihydroquinolin-2 ( lü )-o n e , substance to be examined and the reference substance
G. R = O H , R ' = H: 5-[(2RS)-2,3-dihydroxypropoxy]-3,4- separately in 2-propanol R, evaporate to dryness and record
dihydroquinolin-2 ( 1H)-one, new spectra using the residues.
I. R = N H -C (C H 3)3, R ' = B n 7-brom o-5-[(2i?5)-3-[(l,l- TESTS
(dimethylethyl) amino] -2-hydroxypropoxy]-3,4-
Related substances
dihydroquinolin-2 ( 1if)-o n e, L iquid chromatography (2.2.29).
Test solution Dissolve 25 m g of the substance to be examined
in the mobile phase and dilute to 25.0 m L w ith the mobile
phase.
Reference solution (a) Dilute 1.0 m L of the test solution to
100.0 m L with the mobile phase. Dilute 1.0 m L o f this
solution to 10.0 m L with the mobile phase.
E. 5 ,5 '-[(2-hydroxypropan-l,3-diyl)bis(oxy)]bis(3,4- Reference solution (b) Dissolve 5 mg o f carvedüol
dihydroquinolin-2 (l//)-o n e )3 impurity C CRS in 5.0 m L o f the mobile phase and dilute to
100.0 m L with the mobile phase. Dilute 4.0 m L o f the
solution to 100.0 m L with the mobile phase. D ilute 1.0 m L
hv 0H h o f this solution to 10.0 m L with the mobile phase.
O N CH 3 and enantiomer
Reference solution (c) Dissolve 5 mg o f carvedüolfor system
H3C ch3
sttitabüày CRS (containing impurities A and D ) in the mobile
phase and dilute to 50.0 m L with the mobile phase.
H . 5 -[ (2RS)-3-[( 1,1 -dimethylethyl)amino] -2- Column:
hydroxypropoxy] quinolin-2 (1H) -one. — size. I = 0.150 m , 0 = 4.6 mm;
— stationary phase, end-capped octylsüyl süica gel for
____________________________________________________________________________________________ PhEur
chromatography R (5 |im );
— temperature: 55 °C.
Mobile phase Dissolve 1.77 g of potassium dihydrogen
phosphate R in water R and dilute to 650 m L w ith the same
★ ★
Carvedilol ★ ★ solvent; adjust to p H 2.0 with phosphoric add R and add
350 m L o f acetonitrUe R.
(Ph. Eur. monograph 1745)
Flow rate 1.0 mL/min.
H OH Detection Spectrophotom eter at 240 nm .
Injection 20 |iL.
Run time 6 times the retention rime o f carvedilol.
Identification of impurities U se the chrom atogram supplied
with carvedüolfor system suitabüùy CRS and the
chrom atogram obtained with reference solution (c) to identify
the peaks due to impurities A and D ; use the chrom atogram
C24H 26N 20 4 406.5 72956-09-3 obtained with reference solution (b) to identify the peak due
to im purity C.
A c tio n a n d u s e
Relative retention W ith reference to carvedilol (retention
Beta-adrenoceptor antagonist; arteriolar vasodilator.
tim e = about 4 m in): impurity A = about 0.5;
P r e p a r a tio n im purity C = about 2.9; im purity D = about 3.8.
Carvedilol Tablets System suitability:
P h E u r ____________________________________________________________________________________________
— resolution: minimum 3.5 between the peaks due to
im purity A and carvedilol in the chrom atogram obtained
D E F IN IT IO N w ith reference solution (c);
(2RS ) - 1-(9/Jr-Carbazol-4-yloxy)-3- [ [2- — signal-to-noise ratio: minimum 10 for the peak due to
(2-methoxyphenoxy)ethyl]amino]propan-2-ol. im purity C in the chromatogram obtained w ith reference
C o n te n t solution (b).
99.0 per cent to 101.0 per cent (dried substance). Limits:
— correction factor, for the calculation o f content, multiply the
CHARACTERS
peak area of impurity A by 2.0;
A p p e a ra n c e
W hite or alm ost white, crystalline powder.
2016 Castor Oil 1-429
P h E u r ____________________________________________________
D E F IN IT IO N
Fatty oil obtained by hydrogenation of Virgin Castor
oil (0051). It consists mainly of th e triglyceride of
12-hydroxystearic (12-hydroxyoctadecanoic) acid.
CHARACTERS
A p p e a ra n c e
Fine, almost white or pale yellow powder or almost white or
pale yellow masses or flakes.
A. 1- [ [9- [2-hydroxy-3- [[2-
(2-methoxyphenoxy)ethyI] aminojpropyl] -9if-carbazol-4- S o lu b ility
yl] oxy] -3- [[2-(2-methoxyphenoxy) ethyl] amino]propan-2-ol, Practically insoluble in water, slightly soluble in methylene
chloride, very slightly soluble in anhydrous ethanol,
practically insoluble in light petroleum.
ID E N T IF IC A T IO N
A. M elting point (2.2.14): 83 °C to 88 °C.
B. Hydroxyl value (see Tests).
C. Composition o f fatty adds (see
1-430 Castor Oil 2016
boil under a reflux condenser for 30 min. Allow to cool. R e sid u a l eth y len e oxide a n d d io x an (2.4.25)
Acidify the solution with 20 m L of hydrochloric acid R. Shake M aximum 1 ppm of residual ethylene oxide and 10 ppm of
the mixture with 50 m L of ether R and allow to stand until residual dioxan.
separation of the layers is obtained. Transfer the clear u p p er H eav y m e ta ls (2.4.8)
layer to a suitable tube, add 5 g of anhydrous sodium sulfate R, M aximum 10 ppm .
close the tube and allow to stand for 30 min. Filter and
12 m L o f solution S, filtered if necessary, complies with
evaporate the filtrate to dryness on a water-bath. Dissolve
test A. Prepare die reference solution using lead standard
50 m g of the residue in 25 m L of ether R.
solution (1 ppm Pb) R.
Reference solution Dissolve 50 m g o f ricindeic acid R in
W a te r (2.5.12)
methylene chloride R and dilute to 25 m L with the same
M aximum 3.0 per cent, determined on 2.000 g.
solvent.
Plate TLC octadecylsUyl silica gel plate R. T o ta l a sh (2.4.16)
M aximum 0.3 per cent, determ ined on 2.0 g.
Mobile phase methylene chloride R, glacial acetic acid R,
acetone R (10:40:50 VIVIV). STORA GE
Application 2 pL. Protected from light.
Development Over a path of 8 cm. L A B E LL IN G
Drying In a current of cold air. The label states:
Detection Spray w ith an 80 g/L solution of phosphomoJybdic — the am ount o f ethylene oxide reacted with castor oil
add R in 2-propanol R and heat at 120 °C for 1-2 min. (nominal value),
— where applicable, that the substance is suitable for use in
Restdts T he principal spot in the chromatogram obtained with
the manufacture of parenteral preparations.
the test solution is similar in position and colour to the
principal spot in the chromatogram obtained with the ______________________________________________________________ PhEur
reference solution.
D . Place about 2 g o f the substance to be examined in a test-
tube and add 0.2 m L of sulfuric add R. Close the tube using
a stopper fitted with a glass tube bent twice at right angles.
H eat the tube until white fumes appear. Collect the fumes in
Hydrogenated Polyoxyl Castor Oil *****
*. *
1 m L of mercuric chloride solution R. A white precipitate is (Macrogolgfycerol Hydroxystearate, *
formed and the fumes turn a filter paper impregnated with Ph Eur monograph 1083)
alkaline potassium tetraiodomercurate solution R black. PhEir__________________________________________________________
TESTS D E F IN IT IO N
S o lu tio n S Contains mainly trihydroxystearyl glycerol ethoxylated with
Dissolve 5.0 g in carbon dioxide-free water R and dilute to 7 to 60 molecules of ethylene oxide (nominal value), with
50 m L with the same solvent. small amounts o f macrogol hydroxystearate and o f the
A p p e a ra n c e o f so lu tio n corresponding free glycols. It results from the reaction of
Solution S is not m ore opalescent than reference hydrogenated castor oil with ethylene oxide.
suspension HI ('2.2.1) and not more intensely coloured th an
CHA RACTERS
reference solution BY5 (2.2.2, Method II). If intended for use
A p p e a ra n c e
in the manufacture o f parenteral preparations, solution S is
— if less than 10 units o f ethylene oxide p er molecule:
not m ore intensely coloured than reference solution BY6
yellowish, turbid, viscous liquid;
(2.2.2, Method II).
— if m ore than 20 units of ethylene oxide p er molecule:
A lk alin ity white or yellowish semi-liquid o r pasty mass.
Dissolve 2.0 g in a hot mixture of 10 m L o f water R and
S o lu b ility
10 m L o f ethanol (96 per cent) R. Add 0.1 m L of bromothymol
— if less than 10 units o f ethylene oxide per molecule:
blue sohaton R l. N o t more than 0.5 m L of 0.1 M hydrochloric
practically insoluble in water, soluble in acetone,
add is required to change the colour of the indicator to
dispersible in ethanol (96 p er cent);
yellow.
— if m ore than 20 units of ethylene oxide per molecule:
A cid v alu e (2.5.1) freely soluble in water, in acetone and in ethanol
M axim um 2.0, determined on 5.0 g. (96 per cent), practically insoluble in light petroleum.
H y d ro x y l v alu e (2.5.3, Method A) ID E N T IF IC A T IO N
See T able 1082.-1.
A. Iodine value (see Tests).
Io d in e v alu e (2.5.4) B. Saponification value (see Tests).
25 to 35.
C. Thin-layer chromatography (2.2.27).
S a p o n ific a tio n v a lu e (2.5.6)
Test sducion T o 1 g o f the substance to be examined, add
See T able 1082.-1.
100 m L o f a 100 g/L solution of potassium hydroxide R and
boil under a reflux condenser for 30 min. Allow to cool.
Table 1082.-1 Acidify the solution with 20 m L o f hydrochloric add R. Shake
Ethylene oxide units per Hydroxyl value Saponification value the mixture with 50 m L of ether R and allow to stand until
nxdcculg (nominal valn»|_________________________________ separation o f the layers is obtained. Transfer the clear upper
30 - 35 65 - 82 60 - 75 layer to a suitable tube, add 5 g o f anhydrous sodium sulfate R,
50 48 - 68 38 - 52
• close the tube and allow to stand for 30 min. Filter and
1-432 Castor Oil 2016
7 0 -9 0 7 0 -9 0
C. Composition of fatty ad d s (see Tests).
25
40 6 0 -8 0 4 5 -6 9 TESTS
A p p e a ra n c e
60 4 5 -6 7 40 -5 1
T h e substance to be examined is clear (2.2.1) and n o t more
intensdy coloured (2.2.2, Method II) than 20 m L of a
mixture o f 0.25 m L of blue primary solution, 0.25 m L o f red
primary solution, 0.8 m l. o f yellow primary solution, and
2016 Castor Oil 1-433
18.7 m L of a solution prepared by diluting 4.0 m L of Calculate the percentage content o f each fatty a d d by the
hydrochloric add R1 to 100.0 m L with water R. normalisation procedure.
O p tic a l ro ta tio n (2.2.7) C o rre a the area of the peak due to methyl ridnoleate, by
+ 3.5° to + 6.0°. multiplying by a factor R calculated using the following
expression:
Specific a b so rb a n c e (2.2.25)
Greater than 0.7 and mayi'mum 1.5, determined at the
m\ x A2
absorption maximum at 270 nm.
A\ x m 2
T o 1.00 g add ethand (96 per cent) R and dilute to 100.0 m L
with the same solvent.
m\ = mass o f methyl ridnoleate in the reference solution;
A d d v alu e (2.5.1) m2 = mass o f methyl stearate in the reference solution;
M aximum 0.8. A 1 = area o f the peak due to methyl ridnoleate in the
Dissolve 5.00 g in 25 m L of the prescribed mixture o f chromatogram obtained with the reference solution;
solvents. A 2 = area o f the peak due to methyl stearate in the
H ydroxyl v alu e (2.5.3, Method A) chromatogram obtained with the reference solution.
M inim um 160. Composition of the fatty-acid fraction of the oU:
P e ro x id e v alu e (2.5.5, Method A) — palmitic add: maximum 2.0 p e r cent;
M aximum 5.0. — stearic add: maximum 2.5 per cent;
— oleic add: 2.5 per cent to 6.0 p er cent;
U n sap o n ifiab le . m a tte r (2.5.7)
— linoleic add: 2.5 per cent to 7.0 per cent;
M aximum 0.8 per cent, determined on 5.0 g. — linolenic add. maximum 1.0 p er cent;
O il o b ta in e d b y e x tra c tio n a n d a d u lte ra tio n — eicosenoic add. maximum 1.0 p er cent;
In a ground-glass-stoppered tube about 125 m m long and — ridnoleic acid. 85.0 per cent to 92.0 per cent;
18 mm in internal diameter, thoroughly mix 3 m L o f the — any other fatty add: maximum 1.0 per cent.
substance to be examined with 3 m L o f carbon disulfide R.
W a te r (2.5.32)
Shake for 3 min with 1 m L of sulfuric add R. T h e mixture is
M aximum 0.3 per cent, or maximum 0.2 per cent if intended
less intensely coloured than a freshly prepared mixture of
for use in the manufacture of parenteral preparations,
3.2 m L o f ferric chloride solution R l, 2.3 m L o f water R and
determined on 1.00 g.
0.5 m L o f dilute ammonia R l.
STORAGE
C o m p o sitio n o f fa tty a d d s
In an airtight, well-filled container, protected from light.
Gas chromatography (2.4.22) with the following
modifications. L A B E L L IN G
Use the mixture o f calibrating substances in T able 2.4.22.-3. T he la b d states, where applicable, that the substance is
Test solution Introduce 75 mg o f the substance to be suitable for use in the manufacture of parenteral
examined into a 10 m L centrifuge tube with a screw cap. preparations.
Dissolve in 2 m L o f 1,1-dimeihylethyl methyl ether R l with -----------------------------------------------------------------------------PhEur
shaking and heat gently (50-60 °C). T o the still-warm
solution, add 1 m L o f a 12 g/L solution of sodium R in
anhydrous methanol R, prepared with the necessary
precautions, and shake vigorously for at least 5 min. Virgin Castor Oil *****
Add 5 m L o f distilled water R and shake vigorously for about
30 s. Centrifuge for 15 min at 1500 g. Use the u p p er layer. C astor Oil ****
Reference solution Dissolve 50 mg of methyl ridnoleate CRS and (Ph. Eur. monograph 0051)
50 mg o f methyl stearate CRS in 10.0 m L o f 1,1-dimethylethyl
methyl ether R l. A c tio n a n d u se
Stim ulant laxative; emollient.
Column.:
— material', fused silica; P re p a r a tio n
— sizer. I = 30 m , 0 = 0.25 mm; Zinc and Castor Oil Ointment
— stationary phase: macrogol 20 000 R (film thickness
PhEur__________________________________________________
0.25 jim).
Carrier gas helium for chromatography R. D E F IN IT IO N
Fatty oil obtained by cold expression from the seeds of
Flow rate 0.9 m l An in.
Ridnus communis L. A suitable antioxidant may be added.
Split ratio 1:100.
P R O D U C T IO N
Temperature:
D uring the expression step, the tem perature of the oil m ust
not exceed 50 °C.
Time Temperature
(min) CC) CHARACTERS
Column 0 -5 5 215 A p p e a ra n c e
Injection port 250
Clear at 40 °C, slightly yellow, viscous, hygroscopic liquid.
S o lu b ility
Detector 250
Slightly soluble in light petroleum, m isdble with ethanol
(96 per cent) and with gladal acetic ad d .
Detection Flam e ionisation. R elativ e d e n sity
Injection 1 jiL . About 0.958.
1-434 Cefaclor 2016
Refractive index About 1.479. Calculate the percentage content o f each fatty a d d by the
normalisation procedure.
ID E N T IF IC A T IO N
First identification B, C. C orrect the area o f the peak due to methyl ridnoleate, by
multiplying by a factor R calculated using the following
Second identification A , B.
expression:
A. A mixture o f 2 m l. o f the substance to be examined and
8 m l. o f ethanol (96 per cent) R is d e a r (2.2.1). m i x A?
B. Specific absorbance (see Tests). A \ x m?
C. Com position of fatty a d d s (see Tests).
mi = mass o f methyl ridnoleate in the reference solution;
TESTS
m2 = mass o f methyl stearate in the reference solution;
O p tic a l ro ta tio n (2.2 . 7)
A i = area o f the peak due to methyl ridnoleate in the
+ 3.5° to + 6.0°.
chrom atogram obtained with the reference solution;
S p ecific a b so rb a n c e (2.2.25) A 2 = area o f the peak due to methyl stearate in the
M aximum 0.7, determ ined at the absorption m aximum at chrom atogram obtained with th e reference solution.
270 nm .
Composition of the fatty-acid fraction of the oik
T o 1.00 g add ethanol (96 per cent) R and dihite to 100.0 m l- — palmitic add: maximum 2.0 per cent;
with the same solvent. — stearic add: maxim um 2.5 per cent;
A cid v alu e (2.5.1) — oleic acid : 2.5 p er cent to 6.0 per cent;
M axim um 1.5. — linoleic add : 2.5 per cent to 7.0 per cent;
Dissolve 5.00 g in 25 m L o f the prescribed mixture of — Itnolenic acid : maximum 1.0 per cent;
solvents. — eicosenoic a d d : maximum 1.0 per cent;
— ridnoleic a d d : 85.0 per cent to 92.0 p er cent;
H y d ro x y l v a lu e (2.5.3, Method A)
— any otherfatty add: maximum 1.0 per cent.
M inim um 160.
W a te r (2.5.32)
P e ro x id e v alu e (2.5.5, Method A)
Maximum 0.3 per cent, determined on 1.00 g.
M axim um 10.0.
U n sa p o n ifia b le m a t t e r (2.5.7) STORAGE
M aximum 0.8 per cent, determ ined on 5.0 g. In an airtight, well-filled container, protected from light.
S olubility — disregard limit. 0.1 times the area of the principal peak in
Slightly soluble in water, practically insoluble in methanol the chromatogram obtained with reference solution (b)
and in methylene chloride. (0.1 per cent).
ID E N T IF IC A T IO N H eav y m e ta ls (2.4.8)
Infrared absorption spectrophotometry (2.2.24). M aximum 30 ppm.
Comparison cefaclor CRS. 1.0 g complies with test C. Prepare the reference solution
using 3 m L o f lead standard solution (10 ppm Pb) R.
TESTS
p H (2.2.3) W a te r (2.5.12)
3.0 to 4.5. 3.0 per cent to 6.5 per cent, determined on 0.200 g.
Suspend 0.250 g in carbon dioxide-free water R and dilute to ASSAY
10 m L with the same solvent. Liquid chromatography (2.2.29).
Specific o p tic a l ro ta tio n (2.2.7) Test solution Dissolve 15.0 mg of the substance to be
+ 101 t o + 111 (anhydrous substance). examined in the mobile phase and dilute to 50.0 m L w ith
the mobile phase.
Dissolve 0.250 g in a 10 g/L solution of hydrochloric acid R
and dilute to 25.0 m L with the same solution. Reference solution (a) Dissolve 15.0 mg o f cefaclor CRS in the
mobile phase and dilute to 50.0 m l. with the mobile phase.
R e la te d su b sta n c e s
Liquid chromatography (2.2.29). Reference solution (b) Dissolve 3.0 mg of cefaclor CRS and
3.0 mg o f deka-3-cefaclor CRS (impurity D) in the mobile
Test solution Dissolve 50.0 mg of the substance to be
phase and dilute to 10.0 m L with the mobile phase.
examined in 10.0 m L o f a 2.7 g/L solution of sodium
dihydrogen phosphate R adjusted to p H 2.5 with phosphoric Column:
add R. — sizer. I = 0.25 m, 0 = 4.6 mm;
— stationary phase: octadecylstlyl silica gel for chromatography R
Reference solution (a) Dissolve 2.5 m g of cefaclor CRS and
(5 ^m).
5.0 mg of delta-3-cefaclor CRS (impurity D ) in 100.0 m L o f a
2.7 g/L solution of sodium dihydrogen phosphate R adjusted to Mobile phase Add 220 m L of methanol R to a mixture of
pH 2.5 with phosphoric add R. 780 m L of water R, 10 m L of triethylamine R and 1 g of
sodium pentanesulfonate R, then adjust to p H 2.5 with
Reference solution (b) Dilute 1.0 m L of the test solution to
phosphoric add R.
100.0 m L with a 2.7 g/L solution o f sodium dihydrogen
phosphate R adjusted to p H 2.5 with phosphoric add R. Flow rate 1.5 mlVmin.
Column: Detection Spectrophotometer at 265 nm.
— size: I = 0.25 m, 0 = 4.6 mm; Injection 20 (iL.
— stationary phase: end-capped octadecylsüyl silica gel for System suitability".
chromatography R (5 (im). — resolution: minimum 2.5 between the peaks due to cefaclor
Mobile phase: and impurity D in the chromatogram obtained with
— mobile phase A: 7.8 g/L solution o f sodium dihydrogen reference solution (b); if necessary, adjust the
phosphate R adjusted to p H 4.0 with phosphoric add R] concentration of methanol in the mobile phase;
— mobile phase B: mix 450 m L of acetonitrile R with 550 m L — symmetry factor, maximum 1.5 for the peak due to cefaclor
of mobile phase A; in the chromatogram obtained with reference, solution (b);
— repeatability: maximum relative standard deviation of
1.0 per cent after 6 injections of reference solution (a).
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V) IM P U R IT IE S
0 -3 0 9 5 -»75 5 -»25
3 0 -4 5 75 -» 0 25 -» 100
4 5 -5 5 0 100
c o 2h
H NH2
C. (6i?,7i?)-7-[[(25)-2-ainino-2-phenylacetyl]amino]-3- H20
chloro-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic
H H
acid,
W a te r (2.5.12)
4.0 per cent to 6.0 per cent, determined on 0.200 g.
Q ’t t y
S u lfa te d ash (2.4.14)
M axim um 0.5 per cent, determined on 1.0 g.
D . (6Æ,7£)-7-[[(2S)-2-amino-2-
A SSAY (4-hy droxyphenyl) acetyl] amino] -3-methyl-8-oxo-5-thia-1-
Liquid chromatography (2.2.29). azabicyclo [4.2.0] oct-2-ene-2-carboxylic a d d (L-cefadroxil),
Test solution Dissolve 50.0 m g o f the substance to be
examined in the mobile phase and dilute to 100.0 m l. with H ,n h 2
the mobile phase.
Reference solution (a) Dissolve 50.0 mg of cefadroxil CRS in and enantiomer
r y i h o
the mobile phase and dilute to 100.0 m L with the mobile
phase. U
X J HH
Reference solution (b) Dissolve 5 mg of cefadroxil CRS and
50 m g o f amoxicillin trihydrate CRS in the mobile phase and .E. (6ÆS)-3-(aminomethylene)-6-
dilute to 100 m L with the mobile phase. (4-hydroxyphenyl)piperazine-2,5-dione,
1-438 Cefalexin Monohydrate 2016
ID E N T IF IC A T IO N
Infrared absorption spectrophotom etry (2.2.24).
Comparison cefalexin monohydrate CRS.
TESTS
p H (2.2.3)
4.0 to 5.5.
F. (6R,7R)-7- [ [(2R)-2- [ [(2.&S)-2-ammo-2- Dissolve 50 mg in carbon dioxide-free water R and dilute to
(4-hydroxyphenyl) acetyl] amino] -2- 10 m L with the same solvent.
(4-hydroxyphenyI) acetyi] amino]-3-methyl-8-oxo-5-thia-1- S p ecific o p tic a l ro ta tio n (2.2.7)
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, + 149 to + 158 (anhydrous substance).
HO
Dissolve 0.125 g in phthalate buffer solution pH 4.4 R and
dilute to 25.0 m L with the same solvent.
R e la te d su b s ta n c e s
Liquid chromatography (2.2.29).
Test solution Dissolve 50.0 m g o f the substance to be
G . 3-hydroxy-4-methylthiophen-2 (5/f)-one,
examined in mobile phase A and dilute to 50.0 m L with
CO,H m obile phase A.
Reference solution (a) Dissolve 10.0 m g o f D-phenylglycine R in
mobile phase A and dilute to 10.0 m L with mobile phase A.
Reference solution (b) Dissolve 10.0 m g o f
0 7-armnodesacetoocycephalosporanic acid CRS in phosphate buffer
solution pH 7.0 R5 and dilute to 10.0 m L with mobile
H . ( 6Æ, 7i?) -7 - [(2,2-dimethylpropanoyl) amino] -3-methyl-8- phase A.
oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid Reference solution (c) Dilute 1.0 m L of reference solution (a)
(7-AD CA pival amide). and 1.0 m L o f reference solution (b) to 100.0 m L with
__________ ^________________________________________________ PhEur mobile phase A.
Reference solution (d) Dissolve 10 m g o f dimethylformamide R
and 10 m g of dimethylacetamide R in mobile phase A and
dilute to 10.0 m L with mobile phase A. D ilute 1.0 m L of
Cefalexin Monohydrate ***** this solution to 100.0 m L with mobile phase A.
★, + Reference solution (e) Dilute 1.0 m L o f reference solution (c)
(Ph. Eur. monograph 0708) * to 20.0 m L with mobile phase A.
CO,H Reference solution (f) Dissolve 10 m g o f cefotaxime sodium CRS
in mobile phase A and dilute to 10.0 m L w ith mobile
phase A. T o 1.0 m L of this solution add 1.0 m L o f the test
solution and dilute to 100 m L with mobile phase A.
Column:
— size: I = 0.10 m , 0 = 4.6 mm ;
— stationary phase, spherical octadecylsüyl süica gel for
ClöH 17N 30 4S,H20 365.4 23325-78-2 chromatography R (5 pm).
A c tio n a n d u se Mobile phase:
— mobile phase A: phosphate buffer solution pH 5.0 R;
C ephalosporin antibacterial.
— mobile phase B: methanol R2~,
P r e p a r a tio n s
Cefalexin Capsules
Time Mobile phase A Mobile phaae B
Cefalexin Oral Suspension
(min) (per cent V/V) (per cent V/V)
Cefalexin Tablets 0 -1 98 2
PhEur_______________________________________________ 1 -20 98->70 2->30
D E F IN IT IO N
(6i?,7i?)-7-[[(2iQ-2-Amino-2-phenylacetyl]amino]-3-methyl- Flow rate 1.5 mL/min.
8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic a d d
m onohydrate.
Detection Spectrophotom eter at 220 nm .
Iiyection 20 (iL o f the test solution and reference
Semi-synthetic product derived from a fermentation product.
solutions (c), (d), (e) and (f).
C o n te n t
System suitability:
95.0 per cent to 102.0 per cent (anhydrous substance).
— resolution: minimum 2.0 betw een the peaks due to
CHA RACTERS impurities A and B in the chrom atogram obtained with
A p p e a ra n c e reference solution (c) and minimum 1.5 betw een the
W hite or alm ost white, crystalline powder. peaks due to cefalexin and cefotaxime in the
S o lu b ility chrom atogram obtained with reference solution (f).
Sparingly soluble in water, practically insoluble in ethanol
(96 p er cent).
2016 Cefalotin Sodium 1-439
Limits:
— impurity B: n o t more than the area o f the 2nd peak in the
chromatogram obtained with reference solution (c)
(1.0 per cent);
— any other impurity, not more than the area o f the 1st peak
in the chromatogram obtained with reference solution (c)
(1.0 per cent);
— total: not m ore than 3 times the area o f the 1st peak in the C. (6£,7i?)-7-[[(2J?)-2-[[(2£)-2-amino-2-
chrom atogram obtained with reference solution (c) phenylacetyl] amino] -2-phenjdacetyl] amino]-3-methyl-8-oxo-
(3.0 per cent); 5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic ad d ,
— disregard limit, the area of the 2nd peak in the
chrom atogram obtained with reference solution (e)
(0.05 per cent); disregard any peaks due to
dimethylformamide or dimethylacetamide. h3c oh
(5 pm).
Mobile phase methanol R, acewnitrile R, 13.6 g/L solution of
potassium dihydrogen phosphate R, water R
(2:5:10:83 VIV/V/V).
Cefalotin Sodium ★
★ ★
*
Flow rate 1.5 mL/xnin. ★, .*
Detection Spectrophotom eter at 254 nm. (Ph. Ever, monograph 0987) *★ *
Injection 20 pL.
C02Na O
System suitability: reference solution (b):
— resolution: minim um 4.0 between the peaks due to
cefalexin an d cefradine.
Calculate the percentage content o f cefalexin monohydrate.
STORA GE
Protected from light. Ci6H15N2Na06S2 418.4 58-71-9
IM P U R IT IE S
Action and use
H NHj Cephalosporin antibacterial.
COjH
PhEtr.
O * D E F IN IT IO N
A. (2i?)-2-amino-2-phenylacetic add (D-phenylglycine), Sodium (6R,lR)-3- [(acetyloxy)methyl]-8-oxo-7- [(thiophen-2-
ylacetyl) amino] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-
ayn
carboxylate.
V h' V ^ Semi-synthetic product derived from a fermentation p ro d u ct
C o n te n t
H H 96.0 per cent to 102.0 per cent (anhydrous substance).
B. (6i?,7J?)-7-amino-3-methyl-8-oxo-5-thia-1- CHARACTERS
azabicydo[4.2.0]oct-2-ene-2-carboxylic A p p e a ra n c e
add(7-aminodesacetoxycephalosporanic ad d , 7-ADCA), W hite or almost white powder.
1-440 Cefalotin Sodium 2016
impurities for demonstration o f compliance. See also 5.10. yl) sulfanyl] methyl] -8-oxo-5-thia-1-azabicydo [4.2.0] oct-2-ene-
Control of impurities in substances for pharmaceutical use): A , C. 2-carboxylate (cefamandole sodium), with sodium carbonate.
co 2h Semi-synthetic product derived from a fermentation p ro d u ct
Content
— cefamandole nafate (C 19H i7N $N a06S2): 93.0 per cent to
102.0 per cent (anhydrous and sodium carbonate-free
H H
c n r substance), for the sum of the content of cefamandole
nafate and cefamandole sodium expressed as cefamandole
A. (6R,7i?)-3-methyl-8-oxo-7- [(thiophen-2-ylacetyl)amino]-5- nafate;
thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic a d d — cefamandole sodium (Q gH nN gN aO sSj): maximum
(deacetoxycefalotin), 10.0 per cent (anhydrous and sodium carbonate-free
CO2H substance);
— sodium carbonate (Na2C 0 3): 4.8 per cent to 6.4 per cent.
CHARACTERS
cn ^ s Appearance
W hite or almost white powder.
B . (6R, 7i?)-3-(hydroxymethyI)-8-oxo-7- [(thiophen-2- Solubility
ylacetyl)amino]-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2- Freely soluble in water, sparingly soluble in methanol.
caiboxylic a d d (deacetylcefalotin) , IDENTIFICATION
CO2H 0 A. Infrared absorption spectrophotometry (2.2.24).
Comparison cefamandole nafate CRS.
B. It gives reaction (a) of sodium (2.3.1).
H H S TESTS
Solution S
C. ( 6R,7i?)-3-[(acetyloxy)methyI]-7-amino-8-oxo-5-thia-1-
Dissolve 2.5 g in carbon dioxide-free water R and dilute to
azabicyclo[4.2.0]oct-2-ene-2-carboxylic a d d (7-ACA),
25 m L with the same solvent.
Appearance o f solution
Solution S is clear (2.2.1) and its absorbance (2.2.25) at
475 nm is not greater than 0.03.
pH
6.0 to 8.0 for solution S, measured after 30 min.
Specific optical rotation (2.2.7)
D . (5aR,6Æ)-6- [(thiophen-2-ylacetyl)amino]-5a,6-dihydro-
—35.0 to —45.0 (anhydrous and sodium carbonate-free
3/fj7H -azeto [2,1-b] furo [3,4-d] [1,3] thiazine-1,7 (4ü)-dione
substance).
(cefalotin lactone).
Dissolve 1.00 g in acetate buffer solution pH 4 7 R1 and dilute
Ph Eur
to 10.0 m L with the same solvent.
Related substances
Liquid chromatography (2.2.29). Prepare the solutions
***** immediately before use.
Cefamandole Nafate ★ ★
Solvent mixture M ix 18 volumes of acetonitrile R and
(Ph. Eur. monograph 1402) ***** 75 volumes of a 10 per cent V/V solution of triethylantine R
COjNa previously adjusted to p H 2.5 with phosphoric add R.
Test solution Dissolve 0.100 g o f the substance to be
V -N ^ V ^ 5 examined in the solvent mixture and dilute to 10.0 m L with
the solvent mixture.
if H H x / Reference solution (a) Dilute 1 mL of the test solution to
1 O N=N
10 m L with the solvent mixture, then heat at 60 °C for
Compound R Molecular Formula
30 min.
Mr
Cefamandole nafate CHO Ci9H17NgNaOeS2 512.5
Reference solution (b) Dilute 1.0 m L of the test solution to
100.0 m L with the solvent mixture.
Cefamandole sodium H C18H17N6NaOsS2 484.5
Column:
— sizer. I = 0.25 m, 0 = 4.6 mm;
A ctio n a n d use — stationary phase: octadecylsHyl silica gelfor chromatography R
Cephalosporin antibacterial. (5 nm).
P h B r ______________________________________
Mobile phase:
— triethylamine phosphate solution: dissolve 2.0 g of sodium
D E F IN IT IO N pentanesulfonate R in 350 m L of water R, add 40 m L of
Mixture o f sodium (6i?,7i?)-7-[[(2J?)-2-(f6rmyloxy)-2- triethylamine R, adjust to p H 2.5 with phosphoric add R
phenylacetyl]amino]-3-[[(l-m ethyl-lii-tetrazol-5- and dilute to 700 m L with water R;
yl) sulfanyl] methyl]-8-oxo-5-thia-l -azabicyclo [4.2.0] oct-2-ene- — mobile phase A: mix 1 volume of the triethylamine
2-carboxylate and sodium (6i?,7i?)-7-[[(2i?)-2-hydroxy-2- phosphate solution and 2 volumes of water R;
phenylacetyl] amino] -3- [ [(1 -methyl- lH-tetrazol-5-
1-442 Cefamandole Nafate 2016
Column:
Cefapirin Sodium — sizer. I = 0.30 m , 0 = 4 mm,
(Ph. Eur. monograph 1650) * — stationary phase: octadecylsUyl silica gelfor chromatography R
(10 nm).
CO^Na O
Mobile phase Mix 80 m l. o f dimetkylformamide R, 4.0 mT. o f
glacial acetic acid R and 20 m L of a 4.5 per cent mJm solution
o f potassium hydroxide R. Dilute to 2 L with water R.
Flow rate 2.0 m U m in.
O
Detection Spectrophotom eter at 254 nm.
C 17H 16N 3N a06S2 445.5 24356-60-3 Injection 20 )xL o f the test solution and reference
solutions (b), (c) and (d).
A c tio n a n d u se Run time Twice the retention time o f cefapirin.
Cephalosporin antibacterial. Relative retention W ith reference to cefapirin (retention
PhEur___________________________ _______________________________ time = about 13 min): impurity B = about 0.3;
impurity C = about 0.5; impurity A = about 0.75.
D E F IN IT IO N System suitability Reference solution (d):
Sodium ( 6R,7R)-3-[(acetyloxy)methyl]-8-oxo-7-[[[(pyridin-4- — resolution: minimum 2.0 between the peaks due to
yl) sulfanyl] acetyl] amino] -5-thia-1-azabicydo [4.2.0] oct-2-ene- cefapirin and impurity A.
2-carboxylate.
Limits:
Semi-synthetic product derived from a fermentation product. — any impurity, for each impurity, not more than the area o f
C o n te n t the principal peak in the chromatogram obtained with
96.0 per cent to 102.0 per cent (anhydrous substance). reference solution (b) (1.0 per cent), and not more than
1 such peak has an area greater than 0.3 times the area of
CHARACTERS
the principal peak in the chromatogram obtained with
A p p e a ra n c e
reference solution (b) (0.3 per cent),
W hite or pale yellow powder.
— total: no t more than twice the area of the prindpal peak in
S o lu b ility the chrom atogram obtained with reference solution (b)
Soluble in water, practically insoluble in methylene chloride. (2.0 per cent),
ID E N T IF IC A T IO N — disregard limit, area o f the principal peak in the
A. Infrared absorption spectrophotometry (2.2.24). chromatogram obtained with reference solution (c)
(0.05 per cent).
Comparison cefapirin sodium CRS.
B. It gives reaction (a) o f sodium (2.3.1).
N,iV-Dimethylaniline (2.426, Method E)
M aximum 20 ppm .
TESTS
2-Ethylhexanoic acid (2.4.28)
A p p e a ra n c e o f so lu tio n
M aximum 0.5 per c e n t
Dissolve 2.0 g in water R and dilute to 10.0 m L with the
same solvent. T he solution is clear (2.2.1). D ilute 5.0 m L to W a te r (2.5.12)
10.0 m L with water R. T he absorbance (2.2.25) o f this M aximum 2.0 per cent, determined on 0.300 g.
solution at 450 n m is no t greater than 0.25. Bacterial endotoxins (2.6.14)
p H (2.2.3) Less than 0.17 IU/mg, if intended for use in the m anufacture
6.5 to 8.5. o f parenteral preparations without a further appropriate
procedure for the removal of bacterial endotoxins.
Dissolve 0.100 g in carbon dioxide-free water R and dilute to
10.0 m L with the same solvent. A SSAY
S p ecific o p tica l ro ta tio n (2.2.7) Liquid chromatography (2.2.29) as described in the test for
+ 150 to + 165 (anhydrous substance). related substances with the following modification.
Dissolve 0.500 g in water R and dilute to 25.0 m L with the Injection T est solution and reference solution (a).
sam e solvent. Calculate the percentage content of C 17H 16N 3 N a 0 6S2.
R e la te d su b sta n c e s STORAGE
Liquid chromatography (2.2.29). Prepare the solutions Protected from light. If the substance is sterile, store in a
immediately before use. sterile, tam per-proof container.
Test solution Dissolve 42 mg o f the substance to be examined IM P U R IT IE S
in the mobile phase and dilute to 200.0 m L with the mobile *Specified impurities A , B, C.
phase.
Reference solution (a) Dissolve 42 mg of cefapirin sodium CRS
in the mobile phase and dilute to 200.0 m L with the mobile
phase.
Reference solution (b) Dilute 1.0 m L o f the test solution to
100.0 m L with the mobile phase.
Reference solution (c) Dilute 1.0 m L o f reference solution (b)
to 20.0 m L with the mobile phase.
Reference solution (d) Mix 1 m L o f the test solution, 8 m L of A. (5ai?, 6K)-6-[ [[(pyridin-4-yl)sulfanyl] acetyl] amino]-5a,6-
the mobile phase and 1 m L o f hydrochloric acid R l. H eat at dihydro-3/i,7if-azeto [2,1 -b\ furo [3,4-d\ [1,3] thiazine-1,7 (4H )-
60 °C for 10 min. dione (deacetylcefapirin lactone),
1-444 Cefatrizine Propylene Glycol 2016
co 2h
Dissolve 0.400 g in 1 M hydrochloric acid and dilute to
20.0 m L with the same add.
H ° ll ] R Propylene glycol
Gas chromatography (2.2.28).
o
Solvent mixture acetone R, water R (20:80 VIV).
B. R = OH: (6i?,7i?)-3-(hydroxymethyl)-8-oxo-7-[[[(pyridin- Internal standard solution Dissolve 1.0 g o f dimethylacetamide R
4-yl)sulfanyl]acetyI]amino]-5-thia-l-azabicyclo[4.2.0]oct-2- in the solvent mixture and dilute to 50.0 m L with the solvent
ene-2-carboxylic a d d (deacetylcefapirin), mixture.
C. R = H: (6i?,7i?)-3-methyl-8-oxo-7-[[[(pyridin-4- Test solution Introduce 0.40 g of the substance to be
yl) sulfanyl] acetyl] am ino]-5-thia-1-azabicydo [4.2.0] oct-2-ene- examined into a ground-glass-stoppered test-tube.
2-carboxylic a d d (deacetoxycefapirin). Add 3.0 m L o f the internal standard solution, 1.0 m L o f the
solvent mixture and 2.0 m L o f hydrochloric acid R. Seal the
___________________________________________________________________________________________ PhEur
test-tube and shake.
Reference solution (a) Dissolve 2.0 g o f propylene glycol R in the
solvent mixture and dilute to 100.0 m L with the solvent
★*★ mixture.
★ ★
Cefatrizine Propylene Glycol ★ ★ Reference solution (b) Introduce into a ground-glass-stoppered
***** test-tube 1.0 m L o f reference solution (a) and 1.0 m L o f the
(Ph. Eur. monograph 1403) internal standard solution.
Column'.
— material: stainless steel;
— size: I — 2 m , 0 = 2 mm;
— stationary phase: ethylvinylbenzene-dwinylbeTizene
copolymer R (150-180 pm).
Carrier gas nitrogen for chromatography R.
H OH Flow rate 30 mL/min.
X ^O H and enantioiDer Temperature:
h3c
— column: 200 °C;
— injection port and detector. 250 °C.
C 18Hi8N60 5S2j(CyigO^n 462.5 (base) Detection R am e ionisation.
Injection 1 pL o f the test solution and reference solution (b).
A c tio n a n d u se
Limit:
Cephalosporin antibacterial.
— propylene glycol: 13.0 per cent to 18.0 per cent.
P h E u r ________________________________________________________ Related substances
D E F IN IT IO N Liquid chromatography (2.2.29).
M ixture of (6i?,7i?)-7-[[(2i?)-2-amino-2- Test solution Dissolve 60.0 mg of the substance to be
(4-hydroxyphenyl) acetyl] amino] -8-oxo-3- [[(1H -1,2,3-triazol- examined in the mobile phase and dilute to 100.0 m L with
4-yl) sulfanyl] methyl] -5-thia-1-azabicydo [4.2.0] oct-2-ene-2- the mobile phase.
carboxylic a d d and propane-1,2-diol in molecular Reference solution (a) Dissolve 60.0 m g o f cefatrizine propylene
proportions o f about 1:1. glycol CRS in the mobile phase and dilute to 100.0 m L with
C o n te n t the mobile phase.
95.0 per cent to 102.0 per cent of C i8H i8N 60 5S2 Reference solution (b) Dissolve 30.0 mg o f cefatrizine
(anhydrous substance). impurity A CRS in buffer solution pH 7.0 R and dilute to
100.0 m L with the same buffer solution.
CHARACTERS
A p p e a ra n c e Reference solution (c) Dilute 0.6 m L o f reference solution (a)
W hite or alm ost white powder. to 100.0 m L with the mobile phase.
S o lu b ility Reference solution (d) Dilute 1.0 m L o f reference solution (b)
Slightly soluble in water, practically insoluble in ethanol to~l 00.0 m L with buffer solution pH 7.0 R.
(96 per cent) and in methylene chloride. Reference solution (e) T o 1.0 m L of reference solution (a) add
1.0 m L o f reference solution (b) and dilute to 10.0 m L with
ID E N T IF IC A T IO N
the mobile phase.
A. Infiared absorption spectrophotometry (2.2.24).
Column:
Comparison cefatrizine propylene glycol CRS. — sizer. I = 0.25 m , 0 = 4 mm;
B. Examine the chromatograms obtained in the test for — stationary phase: octadecylsifyl silica gel for chromatography R
propylene glycol. (5 pm).
Results T he prindpal peak in the chromatogram obtained Mobile phase Mix 5 volumes of acetonitrile R and 95 volumes
w ith the test solution is similar in retention time and size to of a 2.72 g/L solution of potassium dihydrogen phosphate R in
the principal peak in the chrom atogram obtained with water R.
reference solution (b). Flow rate 2 m lA nin.
TESTS Detection Spectrophotometer at 272 nm.
S p ecific o p tic a l ro ta tio n (2.2.7)
+ 63 to + 69 (anhydrous substance).
2016 Cefazolin Sodium 1-445
W a te r (2.5.12) D E F IN IT IO N
M aximum 1.5 per cent, determined on 0.500 g.
Sodium (6R,7R)-3- [ [(5-methyl-1,3,4-thiadiazol-2-
S u lfated a sh (2.4.14) yl) sulfanyi] methyl] -8-oxo-7 - [(lfi-tetrazol-1-ylacetyl) amino] -
M aximum 0.1 per cent, determined on 1.0 g. 5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylate.
A SSAY Semi-synthetic product derived from a fermentation product.
liq u id chromatography (2.2.29) as described in the test for C o n te n t
related substances w ith the following modifications. 95.0 per cent to 102.0 per cent (anhydrous substance).
Injection T est solution and reference solution (a). CHARACTERS
System suitability: reference solution (a): A p p e a ra n c e
— repeatability: maximum relative standard deviation of White or almost white powder, very hygroscopic.
1.0 per cent after 6 injections.
S o lu b ility
Calculate the percentage content o f C 18H 18N 60 5S2 from the Freely soluble in water, very slightly soluble in ethanol
declared content o f C 18H 18N 60 5S 2 in cefatrizine propylene (96 per cent).
glycol CRS.
It shows polymorphism (5.9).
IM P U R IT IE S
ID E N T IF IC A T IO N
Specified impurities A.
A. Infrared absorption spectrophotometry (2.2.24).
CO,H Preparation Dissolve 0.150 g in 5 m L of water R, add 0.5 m L
of dilute acetic acid R, swirl and allow to stand for 10 m in in
iced water. Filter the precipitate and rinse with 1-2 m L of
water R. Dissolve in a mixture of 1 volume of water R and
9 volumes of acetone R. Evaporate the solvent almost to
N-NH dryness, then dry in an oven at 60 °C for 30 m in.
Comparison cefazolin CRS.
A. (6R,7R)-7-am ino-8-oxo-3-[ [(1 1,2,3-triazol-4-
B. It gives reaction (a) o f sodium (2.3.1).
yl) sulfanyi] methyl] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-
carboxylic a d d (7-ACA triazole). TESTS
puEir S o lu tio n S
Dissolve 2.50 g in carbon dioxide-free water R and dilute to
25.0 m L with the same solvent.
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and its absorbance (2.2.25) at
430 nm is not greater than 0.15.
p H (2.2.3)
4.0 to 6.0 for solution S.
S p ecific o p tic a l ro ta tio n (2.2.7)
- 2 4 to - 1 5 (anhydrous substance).
Dissolve 1.25 g in water R and dilute to 25.0 m L with the
same solvent.
A b so rb a n c e (2.2.25)
Dissolve 0.100 g in water R and dilute to 100.0 m L with the
same solvent. Dilute 2.0 mL of the solution to 100.0 m L
with sodium hydrogen carbonate solution R. Examined between
220 nm and 350 nm, the solution shows an absorption
1-446 Cefazolin Sodium 2016
maximum at 272 nm . The specific absorbance at the Flow rate 1.2 mL/min.
maximum is 260 to 300 (anhydrous substance). Detection Spectrophotom eter at 254 nm .
R e la te d su b s ta n c e s Injection 5 )iL.
L iquid chrom atography (2.2.29). System suitability: reference solution (b):
Test solution Dissolve 50.0 m g of the substance to be — resolution: minimum 2.0 between the peaks due to
examined in mobile phase A and dilute to 20.0 m L with cefazolin and impurity L (see Figure 0988.-1).
mobile phase A. Limits:
Reference solution (a) Dilute 1.0 m L of the test solution to — any impurity: for each impurity, no t m ore than the area of
100.0 m L with mobile phase A the principal peak in the chrom atogram obtained with
Reference solution (b) Dissolve 20 m g o f the substance to be reference solution (a) (1.0 p er cent);
examined in 10 m L o f a 2 g/L solution of sodium hydroxide R. — total: not more than 3.5 times the area of the principal
Allow to stand for 15-30 min. Dilute 1.0 m L of the solution peak in the chromatogram obtained with reference
to 20 m l- with mobile phase A. solution (a) (3.5 per cent);
— disregard limit: 0.05 times the area of the principal peak in
Column:
— size. I = 0.125 m , 0 = 4.0 mm; the chromatogram obtained with reference solution (a)
— stationary phase: octadecylsûyl silica gel for chromatography R (0.05 per cent).
(3 nm); A ^ N -D im eth y lan ilin e (2.4.26, Method B)
— temperature: 45 °C. M aximum 20 ppm.
Mobile phase: W a te r (2.5.12)
— mobile phase A: solution containing 14.54 g/L o f disodium M aximum 6.0 per cent, determ ined on 0.300 g.
hydrogen phosphate R and J.53 g/L o f potassium dihydrogen B a c te ria l en d o to x in s (2.6.14)
phosphate R; Less than 0.15 IU/mg, if intended for use in the manufacture
— mobile phase B: acetonitrüe for chromatography R',
of parenteral preparations without a further appropriate
procedure for the removal of bacterial endotoxins.
Time Mobile phase A Mobile phase B
ASSAY
(min) (per cent V/V) (per cent V/V)
Liquid chromatography (2.2.29).
0 -2 98 2
Test solution Dissolve 50.0 mg o f the substance to be
2 -4 98-» 85 2-» 15
examined in the mobile phase and dilute to 50.0 m L with
4 - 10 85-» 60 15 -»40 the mobile phase.
10 - 11.5 60-» 35 40-» 65 Reference solution (a) Dissolve 50.0 m g o f cefazolin CRS in the
mobile phase and dilute to 50.0 m L with the mobile phase.
11J - 12 35 65
Reference solution (b) Dissolve 5.0 m g o f cefuroxime
12- 15 3 5 -»98 6 5 -»2
sodium CRS in 10.0 m L o f reference solution (a) and dilute
15-21 98 2 to 100.0 m L with the mobile phase.
2016 Cefazolin Sodium 1-447
C02H 0
Column.
— size. I = 0.25 m, 0 = 4.6 mm; V -N
— stationary phase: octadecylsQyl silica gel for chromatography R
(5 nm).
Mobile phase M ix 10 volumes of acetomtrUe R and 90 volumes n* n o
i s
of a solution containing 2.77 g/L o f disodium hydrogen
phosphate R and 1.86 g/L o f citric acid R. D . (6R,7R)-3 -[(acetyloxy)methyl] -8-oxo-7- [(lH-tetrazol-1 -
ylacetyl)amino]-5-thia-l-azabicydo[4.2.0]oct-2-ene-2-
Flow rate 1.0 mL/min.
carboxylic ad d ,
Detection Spectrophotom eter at 270 nm.
SH
Injection 20 pL.
System suitability: reference solution (b):
— resolution: minim um 2.0 between the peaks due to
A
} -N
cefazoOn and cefuroxime. H,C
Calculate the percentage content o f cefazolin sodium by
multiplying the percentage content of cefazolin by 1.048. E. 5-m ethyl-1,3,4-thiadiazol-2-thiol (M M TD),
ST O R A G E
In an airtight container, protected from light. If the substance
is sterile, store in a sterile, airtight, tam per-proof container.
IM P U R IT IE S
Other detectable impurities (the following substances would, if " i T s'
V n 0
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or G . (5a_R,6i?)-6- [(1 H-tetrazol-1-ylacetyl) amino] -5a, 6-dihydro-
3H ,7//-azeto [2,1 -¿]furo [3,4-d] [1,3] thiazine-1,7 (4H) -dione,
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these c o 2h 0
impurities for demonstration of compliance. See also 5.10.
°V - N
Control of impurities in substances for pharmaceutical use): A , B,
C, D} E, G} H, I} J, K, L H2N' * r ~ k r
H H
CO2H
H . (6i?,7i?) -3- [(acetyloxy)methyl] -7-amino-8-oxo-5-thia-1-
V _ n^ y ^ s azabicydo[4.2.0]oct-2-ene-2-carboxylic a d d (7-ACA),
S^N CC^H
h h \ ;
H,C n^ Y ^ s
A
s
co^H
II h h \ r
O y=N
H3C
" V f j
« ° tl 7 I
n- n O
t t s
n* n o
C . (6i?,7i?)-3-methyl-8-oxo-7- [(1/7-tetrazol-1- h3c
ylacetyl) amino]-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-
carboxylic a d d , K . (6R,7R)-3- [ [(5-methyl-l ,3,4-thiadiazol-2-
yl) sulfanyi] methyl] -8-oxo-7- [(1/i-tetrazol-1-ylacetyl) amino] -
5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxamide
(cefazolinamide),
1-448 Cefepime Hydrochloride Monohydrate 2016
Im p u rity G
Liquid chromatography (2.2.29). Prepare the solutions
immediatefy before use.
Test solution Dissolve 0.100 g o f the substance to be
examined in 0.01 M nitric acid and dilute to 10.0 m L with
the same ad d .
Reference solution (a) Dilute 0.250 g o f N-methylpyrrolidine R
L. (6Æ,7S)-3-[[(5-methyl-l,3,4-thiadiazol-2- (impurity G) to 100.0 m L with water R. D ilute 2.0 m L of
yl) sulfanyl] methyl] -8-oxo-7 - [( l//-tetrazo l-1-ylacetyl) amino] - this solution to 100.0 m L with 0.01 M nitric acid
5-thia-1-azabicydo [4.2.0] oct-2-ene-2-carboxylic ad d .
Reference solution (b) Dilute 0.250 g o f pyrrolidine R to
___________________________________________________________________________________________ PhEur
100 m L w ith 0.01 M nitric add D ilute 2 m L o f the solution
to 100 m L with 0.01 M nitric add. M ix 5 m L of this solution
with 5 m L o f reference solution (a).
Column:
Cefepime Hydrochloride ***** — size: I — 0.05 m , 0 = 4.6 mm;
— stationary phase', strong cation-exchange resin R (5 |im ).
Monohydrate ***** Mobile phase Mix 1 volume of acetonitrile R and 100 volumes
(Cefepime Dihydrochloride Monohydrate, o f 0.01 M nitric add, filter through a 0.2 |xm filter.
Ph Eur monograph 2126) Flow rate 1 mL/min.
Detection Conductivity detector.
Irqection 100 jiL.
Run time 1.1 times the retention time of cefepime.
Retention time Cefepime = about 50 min, eluting as a
broadened peak.
System suitability:
— symmetry factor, maximum 2.5 for the peak due to
C 19H 26C12N 60 5S*H 20 571.5 123171-59-5 impurity G in the chromatogram obtained with reference
solution (a);
A ctio n a n d use — repeatability: maximum relative standard deviation of
Cephalosporin antibacterial. 5.0 per cent after 6 injections o f reference solution (a);
— peak-to-vaRey ratio: minimum 3 between the peaks due to
P hE ur ___________________________________________________________________________________________
pyrrolidine and im purity G in the chromatogram obtained
D E F IN IT IO N with reference solution (b).
(6R, 1R)-1- [ [(22)-(2-Aminothiazol-4- Calculate the percentage content of impurity G in the test
yl) (methoxyimino)acetyl] amino] -3- solution using reference solution (a).
[(1 -methylpyirolidinio) methyl] -8-oxo-5-ihia-l - Limit.
azabicydo[4.2.0]oct-2-ene-2-carboxylate dihydrochloride — impurity G: maximum 0.5 per cent.
monohydrate. Semi-synthetic product derived from a
R e la te d su b s ta n c e s
fermentation product.
Liquid chromatography (2.2.29). Prepare the solutions
C o n te n t immediately before use or keep refrigerated at 4-8 °C for not more
97.0 per cent to 102.0 per cent (anhydrous substance). than 12 h.
CHARACTERS Test solution Dissolve 70.0 mg o f the substance to be
A p p e a ra n c e examined in mobile phase A and dilute to 50.0 m L with
W hite or almost white, crystalline powder. mobile phase A Sonicate for 30 s and stir for about 5 min.
S o lu b ility Reference solution (a) Dissolve 70.0 mg of cefepime
Freely soluble in water and in m ethanol, practically insoluble dihydrochloride monohydrate CRS in mobile phase A and dilute
in methylene chloride. to 50.0 m L with mobile phase A. Sonicate for 30 s and stir
for about 5 min.
ID E N T IF IC A T IO N
A. Infrared absorption spectrophotometry (2.2.24). Reference solution (b) Dilute 1.0 m L o f the test solution to
10.0 m L w ith mobile phase A D ilute 2.0 m L of this solution
Comparison cefepime dihydrochloride monohydrate CRS.
to 100.0 m L with mobile phase A.
B. It gives reaction (a) of chlorides (2.3.1).
Reference solution (c) Dissolve 7 mg o f cefepime dihydrochloride
TESTS monohydrate for system suitability CRS (containing impurities
A p p e a ra n c e o f so lu tio n A, B and F ) in mobile phase A and dilute to 5 m L with
T he solution is clear (2.2.1) and not more intensely coloured mobile phase A
than reference solution Y3 (2.2.2, Method II). Reference solution (d) Dissolve 2 mg of cefepime
Dissolve 2.0 g in water R and dilute to 20 m L with the same impurity E CRS in mobile phase A and dilute to 25.0 m L
solvent. with mobile phase A. Dilute 1.0 m L of the solution to
S pecific o p tic a l ro ta tio n (2.2.7) 10.0 m L w ith mobile phase A.
+ 40 to + 45 (anhydrous substance). Column:
— size: I = 0.25 m , 0 = 4.6 mm;
Dissolve 0.250 g in water R and dilute to 25.0 m L with the
— stationary phase: end-capped octadecylsüyl silica gel for
same solvent.
chromatography R (5 pm).
2016 Cefepime Hydrochloride Monohydrate 1-449
3 0 -3 5 50
STO RA G E
50
Protected from lig h t If the substance is sterile, store in a
35 - 36 50-» 100 50 0
sterile, airtight, tam per-proof container.
3 6 -4 5 100 0
IM P U R IT IE S
Specified impurities A, B, E, F, G.
Flow rate 1 mL/min. Other detectable impurities (the following substances would, if
Detection Spectrophotom eter at 254 nm. present at a sufficient level, be detected by one or other o f
the tests in the monograph. They are limited by the general
Injection 10 |iL o f the test solution and reference
acceptance criterion for other/unspecified impurities and/or
solutions (b), (c) and (d).
by the general monograph Substances for pharmaceutical use
Identification of impurities Use the chromatogram supplied (2034). It is therefore n o t necessary to identify these
w ith cefepime dihydrochloride monohydrate for system impurities for demonstration of compliance. See also 5.10.
suitability CRS and the chromatogram obtained with Control of impurities in substances for pharmaceutical use): C, D.
reference solution (c) to identify the peaks due to
impurities A, B and F; use the chromatogram obtained with
reference solution (d) to identify the peak due to impurity E.
Relative retention W ith reference to cefepime (retention
tim e = about 7 min): impurity E = about 0.4;
im purity F = about 0.8; impurity A = about 2.5;
im purity B = about 4.1.
System suitability: reference solution (c):
— resolution: minim um 1.5 between die peaks due to A. (6Ä,7i?)-7-[[(2E)-(2-aminothiazol-4-
impurity F and cefepime. yl) (methoxyimino) acetyl] amino] -3-
Limits: [ ( 1-methylpyrrolidinio) methyl] -8-oxo-5-thia-1-
— correction factors: for the calculation of content, multiply azabicydo[4.2.0]oct-2-ene-2-carboxylate (arm-cefepime),
the peak areas of the following impurities by the
nh2
corresponding correction factor, impurity A = 1.4;
impurity B = 1.4; im purity E = 1.8;
— impurity A: n o t more than 1.5 times the area o f the
principal peak in the chromatogram obtained with
reference solution (b) (0.3 per cent);
— impurities B, F: for each impurity, n o t m ore than the area
o f the principal peak in the chromatogram obtained with
reference solution (b) (0.2 per cent);
— impurity E: n o t m ore than 0.5 times die area of the
B. (6i?,7i^)-7-[[(2Z)-[2-[[(22)-(2-aminothiazol-4-
principal peak in the chromatogram obtained with
yl) (methoxyimino) acetyl] amino] thiazol-4-
reference solution (b) (0.1 per cent);
yl] (methoxyimino)acetyl] amino]-3-
— unspecified impurities: for each impurity, no t m ore than
[(1 -methylpyrrolidmio) methyl] -8-oxo-5-thia-1-
0.5 times the area o f the principal peak in the
. azabicydo [4.2.0] oct-2-ene-2-carboxylate,
chrom atogram obtained with reference solution (b)
(0.10 p er cent);
ch3
— total: n o t m ore than 5 times the area o f the principal peak
in the chrom atogram obtained with reference solution (b) N
(1.0 per cent);
— disregard Hmir. 0.25 times the area o f the principal peak in
the chromatogram obtained with reference solution (b)
(0.05 p er cent).
W a te r (2.5.12) C . (22) -2-(2-aminothiazol-4-yl)-N-(foimylmethyl)-2-
3.0 per cent to 4.5 per cent, determined on 0.400 g. (methoxyimino)acetamide,
1-450 Cefixime 2016
ch 3
C o n te n t
o
nT 95.0 per cent to 102.0 per cent (anhydrous substance).
n^ A co 2h
CHARACTERS
h2n - ^ T
A p p e a ra n c e
W hite or alm ost white, slightly hygroscopic powder.
D. (2Z)-(2-aminothiazol-4-yl)(methoxyimino)acetic ad d , S o lu b ility
Slightly soluble in water, soluble in methanol, sparingly
C02 soluble in anhydrous ethanol, practically insoluble in ethyl
acetate.
1 uyö“ H H
ID E N T IF IC A T IO N
Infrared absorption spectrophotometry (2.2.24).
Comparison cefixime CRS.
E. (6R,7R)-7-am ino-3-[( 1-methylpyrroUdinio)methyl]-8-oxo- If the spectra obtained show differences, dissolve the
5-thia-1-azabicydo [4.2.0] oct-2-ene-2-carboxylate, substance to be examined and the reference substance
separately in methanol R, evaporate to dryness and record
new spectra using the residues.
TESTS
p H (2.2.3)
2.6 to 4.1.
Suspend 0.5 g in carbon dioxide-free water R and dilute to
10 m l. w ith the same solvent.
R e la te d su b s ta n c e s
Liquid chrom atography (2.2.29).
Test solution Dissolve 25.0 mg o f the substance to be
examined in the mobile phase and dilute to 25.0 m L with
F. (6/?,72?)-7-[[[(6.R,7.R)-7-[[(2Z)-(2-aminothiazol-4- the mobile phase.
yl) (methoxyimino) acetyl] amino] -3- Reference solution (a) Dissolve 25.0 mg of cefixime CRS in the
[(1 -methylpyrrolidinio) methyl] -8-oxo-5-thia-1- mobile phase and dilute to 25.0 m l. with the mobile phase.
azabicydo [4.2.0] oct-2-en-2-yl] carbonyl] amino]-3-
Reference solution (b) Dilute 1.0 m L of reference solution (a)
[(1 -methylpyrrolidinio)methyl] -8-oxo-5-thia-1-
to 100.0 m L with the mobile phase.
azabicydo [4.2.0] oct-2-ene-2-carboxylate,
Reference solution (c) Dissolve 10 m g o f cefixime CRS in
ch 3 10 m L of water R. H eat on a water-bath for 45 min and cool
N (in situ preparation o f impurity D ). Inject immediately.
Column:
o — size. I = 0.125 m , 0 = 4 mm;
— stationary phase, octadecylsilyl silica gel for chromatography R
G. 1-methylpyrrolidine (N-methylpyirolidine).
(5 nm);
PhEur — temperature. 40 °C.
— Mobile phase: mix 250 volumes o f acetonitrUe R and
750 volumes o f a tetrabutylammonium hydroxide solution
prepared as follows: dissolve 8.2 g o f tetrabutylammonium
★ ★ hydroxide R in water R and dilute to 800 m L with the
Cefixime ★ ★ same solvent; adjust to p H 6.5 with dilute phosphoric
(Ph. Eur. monograph 1188) *★* acid R and dilute to 1000 m L with water R.
Flow rate 1.0 m l/m in .
,C 0 2h
C02H Detection Spectrophotom eter at 254 nm.
N' ° Injection 10 [iL o f the test solution and reference solutions (b)
, 3 H20 and (c).
Run time 3 times the retention time o f cefixime.
System suitability: reference solution (c):
— resolution: minimum 2.0 between the peaks due to
Cx^xsNjOTS^HaO 507.5 cefixime and im purity D ; if necessary, adjust the
concentration o f acetonitrile in the mobile phase.
Action and use Limits:
Cephalosporin antibacterial. — any impurity, for each impurity, not more than 0.5 times
the area of the prindpal peak in the chromatogram
PhEur__________________________
obtained with reference solution (b) (0.5 per cent);
DEFINITION — total: n o t more than 3 times the area o f the principal peak
(6R,7R)-7 - [ [(2)-2-(2-Aminothiazol-4-yl)-2- in the chromatogram obtained with reference solution (b)
[(carboxymethoxy)imino] acetyl] amino] -3-ethenyl-8-oxo-5- (3 per cent);
thia-1-azabicydo[4.2.0]oct-2-ene-2-carboxylic a d d trihydrate.
Semi-synthetic product derived from a fermentation product.
2016 Cefoperazone Sodium 1-451
ch3 CC^Na o
N "° ch 3
H H S
B. (6 R ,7R)-7-[[(2i?)-2-[[(4-ethyi-2 ,3 -dioxopiperazin-1-
yl)carbonyl] amino] -2-(4-hydroxyphenyl) acetyl] amino] -3-
[(4-methyl-5-thioxo-4j5-dihydro-l//-tetrazol-l-yI)methyl]-8- CjeHigNsNaOTSa 477.4 64485-93-4
oxo-5-thia-l-azabicydo[4.2.0]oct-2-ene-2-carboxylic add, Action and use
Cephalosporin antibacterial.
SH
Preparation
N ^ N '0"» Cefotaxime Injection
N -N
PhEir__________________________________________________________
C. l-methyl-l/f-tetrazole-5-thiol, DEFINITION
Sodium (6i?,7,R)-3-[(acetyloxy)methyl]-7-[[(22)-2-
co 2h (2-aminothiazol-4-yl)-2- (methoxyimino) acetyl] amino] -8-oxo-
5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylate.
Semi-synthetic product derived from a fermentation product.
Content
96.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
D. (6i?,7R)-7-amino-8-oxo-3-[(1H-1,2,3-triazol-4- Appearance
ylsulfanyl) methyl]-5-thia-1-azabicydo [4.2.0] oct-2-ene-2- White or slightly yellow powder, hygroscopic.
caiboxylic add (7-TACA),
Solubility
CO,H o
Freely soluble in water, sparingly soluble in methanol.
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24).
Comparison cefotaxime sodium CRS.
H H
B. It gives reaction (a) of sodium (2.3.1).
E. (6i?,7i?)-3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-l- TESTS
azabicydo[4.2.0]oct-2-ene-2-carboxylic add (7-ACA), Solution S
Dissolve 2.5 g in carbon dioxide-five water R and dilute to
25.0 mL with the same solvent.
Appearance of solution
Solution S is clear (2.2.1). Add 1 mL of glacial acetic add R
to 10 mL of solution S. The solution, examined immediately,
is clear.
pH (2.2.3)
4.5 to 6.5 for solution S.
Specific optical rotation (2.2.7)
+ 58.0 to + 64.0 (anhydrous substance).
F. (6.R,7S)-7-[[(2 R)-2 - [[(4-ethyl-2,3-dioxopiperazine- 1- Dissolve 0.100 g in water R and dilute to 10.0 mL with the
yl) carbonyl] amino] -2-(4-hydroxyphenyl) acetyi] amino]-3- same solvent.
[[(1-methyl-1H-tetrazol-5-yl) sulfanyl] methyl] -8-oxo-5-thia-1- Absorbance (2.2.25)
azabicydo[4.2.0]oct-2-ene-2-carboxylic add. Maximum 0.40 at 430 nm for solution S.
__________________________________________________________ PhEur
Specific absorbance (2.2.25)
360 to 390, determined at the absorption maximum at
235 nm (anhydrous substance).
Dissolve 20.0 mg in water R and dilute to 100.0 mL with the
same solvent. Dilute 10.0 mL of the solution to 100.0 mL
with water R
Related substances
Liquid chromatography (2.2.29). Prepare the solutions
immediately before use.
Solution A Mobile phase B, mobile phase A (14:86 V/V).
1-454 Cefotaxime Sodium 2016
Test solution Dissolve 40.0 mg of the substance to be — total: not more than 3 times the area of the principal peak
examined in solution A and dilute to 50.0 mL with the same in the chromatogram obtained with reference solution (b)
solution. (3.0 per cent);
Reference solution (a) Dissolve 8.0 mg of cefotaxime acid CRS — disregard limit. 0.05 times the area of the principal peak in
in solution A and dilute to 10.0 mL with the same solution. the chromatogram obtained with reference solution (b)
Reference solution (b) Dilute 1.0 mL of the test solution to
(0.05 per cent).
100.0 m l. with solution A. Ethanol (2.4.24System A)
Reference solution (c) Add 1.0 mL of dilute hydrochloric acid R Maximum 1 .0 per cent.
to 4.0 mL of the test solution. Heat the solution at 40 °C for N ,N-Dimethylaniline (2.4.26, Method B)
2 h. Add 5.0 m l. of buffer solution p H 6 .6 R and 1.0 m l. of Maximum 20 ppm.
dilute sodium hydroxide solution R. 2 -Ethylhexanoic a d d (2.4.28)
Reference solution (d) Dissolve 4 mg of cefotaximefor peak Maximum 0.5 per cent m/m.
identification CRS (containing impurities A, B, C, E and F) in W ater (2.5.12)
5 mL of solution A. Maximum 3.0 per cent, determined on 0.300 g.
Column:
Bacterial endotoxins (2.6.14)
— sizer. I = 0.15 m, 0 = 3.9 mm,
Less than 0.05 IU/mg, if intended for use in the manufacture
— stationary phase: octadecylsilyl silica gelfor chromatography R
of parenteral preparations without a further appropriate
(5 nm),
procedure for the removal of bacterial endotoxins.
— temperature: 30 °C.
Mobile phase: ASSAY
— mobile phase A: 7.1 g/L solution of disodium hydrogen Liquid chromatography (2.2.29) as described in the test for
phosphate R adjusted to pH 6.25 using phosphoric acid R; related substances with the following modification.
— mobile phase B: methanol R; Injection Test solution and reference solution (a).
Calculate the percentage content of C 16H i 6N 5Na 0 7 S2 by
Time Mobile phase A Mobile phase B multiplying the percentage content of cefotaxime by 1.048.
(min) (per cent V/V) (per cent V/V) STORAGE
0,- 7 86 14
In an airtight container, protected from light. If the substance
7 -9 86 -»82 1 4 * 18 is sterile, store in a sterile, airtight, tamper-proof container.
9 - 16 82 18 IMPURITIES
1 6 -4 5 82*60 18 * 40 Specified impurities A,B, C, D, E, F
4 5 -5 0 60 40
Other detectable impurities(the following substances would, if
present at a sufficient level, be detected by one or other of
5 0 -5 5 60*86 4 0 * 14 the tests in the monograph. They are limited by the general
55 - 60 86 14 acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these
Flow rate 1 .0 mL/min. impurities for demonstration of compliance. See also 5.10.
Detection Spectrophotometer at 2 3 5 nm. Control of impurities in substances for pharmaceutical use): G.
D . (6/2,72?)-3-[(acetyloxy)methyl]-7-[[(2E)-2-
(2-aminothiazol-4-yl)-2-(methoxyimino)acetyl] amino]-8-oxo-
5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxy!ic add
(jg-cefotaxime),
C 16H 16N 3Na0 7S2 449.4 33564-30-6
DEFINITION
E. (5ai?,6i?)-6-[[(2Z)-2-(2-aiiiinothiazol-4-yl)-2-
(methoxyimino)acetyl]amino]-5a,6-dihydro*3.i/,7i£-azeto[2,l- Sodium (6R,7S)-3-[(carbamoyloxy)methyl]-7-methoxy-8-oxo-
è]furo[3,4-d] [1,3] thiazine-1,7(AH)-dione (deacetylcefotaxime 7- [[(thiophen-2-yl) acetyl] amino]-5-thia-1-
azabicydo [4.2.0] oct-2-ene-2-carboxylate.
lactone),
Semi-synthetic product derived from a fermentation product.
Content
95.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
Appearance
White or almost white, very hygroscopic powder.
Solubility
Very soluble in water, sparingly soluble in ethanol
(96 per cent).
IDENTIFICATION
F. (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2 Z)-2 -[2-[[[(6R,7R)-7- A. Infrared absorption spectrophotometry (2.2.24).
[[(2Z)-2-(2-aminothiazol-4-yl)-2- Comparison cefoxitin sodium CRS.
(metiioxyimino) acetyl] amino]-2-carboxy-8-oxo-5-thia-1- B. It gives reaction (a) of sodium (2.3.1).
azabicydo[4.2.0] oct-2-en-2-yl] methyl] amino] thiazol-4-yl] -2-
(methoxyimino) acetyl] amino] -8-oxo-5-thia-1- TESTS
azabicydo [4.2.0] oct-2 -ene-2 -carboxylic add Solution S
(cefotaxime dimer), Dissolve 2.50 g in carbon dioxide-free water R and dilute to
25 mL with the same solvent.
Appearance of solution
Solution S is clear (2.2.1) and pot more intensely coloured
than intensity 5 of the range of reference solutions of the
most appropriate colour (2.2.2, Method II).
pH (2.2.3)
4.2 to 7.0.
Dilute 2 mL of solution S to 20 mL with carbon dioxide-free
water R.
Specific optical rotation (2.2.7)
+ 206 to + 214 (anhydrous substance).
G. (6R,7R)-3-[(acetyloxy) metiiyl] -7- [[(2 Z)-2 -[2- [[(2 Z)-2- Dissolve 0.250 g in methanol R and dilute to 25.0 mL with
(2-aminothiazol-4-yl)-2- (methoxyimino) acetyl] amino] thiazol- the same solvent.
4-yI] -2-(methoxyimino) acetyl] amino] - 8-oxo-5-thia-1- Related substances
azabicydo [4.2.0] oct-2 -ene-2 -carboxylic add Liquid chromatography (2.2.29). Prepare the solutions
(ATA cefotaxime). immediately before use.
____________________!_________________ PhEur Solution A Dissolve 1.0 g of potassium dihydrogen phosphate R
and 1.8 g of anhydrous disodium hydrogen phosphate R in
1000 mL of water R. To 100 mL of the solution add
800 mL of water R, adjust to pH 7.0 with phosphoric add R
or a 40 g/L solution of sodium hydroxide R and dilute to
1000 mL with water R.
Test solution Dissolve 50 mg of the substance to be examined
in solution A and dilute to 50.0 mL with solution A.
1-456 Cefoxitin Sodium 2016
Reference solution (a) Dilute 1.0 mL of the test solution to — the area of the principal peak in the
disregard limit,
100.0 mL with solution A. chromatogram obtained with reference solution (b)
Reference solution (b) Dilute 1.0 mL of reference solution (a) (0.05 per cent).
to 20.0 mL with solution A. W ater (2.5.12)
Reference solution (c) Dissolve 5 mg of cefoxitin for peak Maximum 1.0 per cent, determined on 0.500 g.
identification CRS (containing impurities A, B, E, H, I and J) Bacterial endotoxins (2.6.14)
in solution A and dilute to 5 mL with solution A. Less than 0.13 IU/mg, if intended for use in the manufacture
Column: of parenteral preparations without a further appropriate
— size. I = 0.25 m, 0 = 4.6 mm; procedure for the removal of bacterial endotoxins.
— stationary phase: phertylsüyl silica gelfor chromatography R
ASSAY
(3.0 pm);
Liquid chromatography (2.2.29).
— temperature. 35 °C.
Test solution Dissolve 25.0 mg of the substance to be
Mobile phase:
examined in water R and dilute to 25.0 mL with the same
— mobile phase A: 1.0 g/L solution of ammoniumformate R
solvent.
adjusted to pH 2.7 with anhydrous formic acid R;
— mobile phase B: acetonitrüe R; Reference solution (a) Dissolve 25.0 mg of cefoxitin
sodium CRS in water R and dilute to 25.0 mL with the same
solvent.
Time Mobile phase A Mobile phase B
Reference solution (b) Dissolve 20.0 mg of 2-(2-thienyl)acetic
(min) (per cent V7V) (per cent V7V)
acid R in water R and dilute to 25.0 mL with the same
0-5 92 8 solvent.
5-50 92->74 8-» 26 Reference solution (c) Mix 1.0 mL of reference solution (a)
5 0-8 5 74 26
and 5.0 mL of reference solution (b).
Column:
— size. I = 0.25 m, 0 = 4.6 mm;
Flow rate 1.0 mL/min. — stationary phase. octadecylsUyl silica gelfor chromatography R
Detection Spectrophotometer at 254 nm. (5 pm).
Injection 20 |xL. Mobile phase acetic acid R, acetomtrUe R, water R
(1:19:81 V/V/V).
Identification of impurities Use the chromatogram supplied
with cefoxitin for peak identification CRS and the Flow rate 1 mL/min.
chromatogram obtained with reference solution (c) to identify Detection Spectrophotometer at 254 nm.
the peaks due to impurities A, B, E, H, I and J. Injection 20 jiL of the test solution and reference solutions (a)
Relative retention With reference to cefoxitin (retention and (c).
time = about 30 min): impurity A = about 0.83; Run time 12 min.
impurity I = about 0.98; impurity H = about 1.06; System suitability: reference solution (c):
impurity E = about 1.11; impurity B = about 1.18; — resolution: m inim u m 3.5 between the 2 principal peaks.
impurity J = about 1.66.
Calculate the percentage content of Ci 6H 16N 3Na0 7 S2 taking
System suitability: reference solution (c):
into account the assigned content of cefoxitin sodium CRS.
— resolution: minimum 2.0 between the peaks due to
impurities H and E; STORAGE
— peak-to-valley ratio: minimum 2.0, where Hp = height In an airtight container. If the substance is sterile, store in a
above the baseline of the peak due to impurity I and sterile, airtight, tamper-proof container.
H v = height above the baseline of the lowest point of the IMPURITIES
curve separating this peak from the peak due to cefoxitin. Specified impurities A,B, E, H, I, J
Limits:
Other detectable impurities(the following substances would, if
— impurity I: not more than the area of the principal peak in present at a sufficient level, be detected by one or other of
the chromatogram obtained with reference solution (a) the tests in the monograph. They are limited by the general
( 1.0 per cent); acceptance criterion for other/unspecified impurities and/or
— impurities E, H: not more than 0.5 times the area of the
by the general monograph Substances for pharmaceutical use
principal peak in the chromatogram obtained with (2034). It is therefore not necessary to identify these
reference solution (a) (0.5 per cent); impurities for demonstration of compliance. See also 5.10.
— impurity J: not more than 0.3 times the area of the
Control of impurities in substancesfor pharmaceutical use): C, D,
principal peak in the chromatogram obtained with F ,G .
reference solution (a) (0.3 per cent);
— impurities A, B: for each impurity, not more than
0.2 times the area of the principal peak in the
chromatogram obtained with reference solution (a)
(0.2 per cent);
— unspecified impurities: for each impurity, not more than
twice the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.10 per cent);
— total: not more than 3 times the area of the principal peak A (6i?,75)-3-(hydroxymethyl)-7-methoxy-8-oxo-7-
in the chromatogram obtained with reference solution (a) [[(thiophen-2-yl)acetyl]amino]-5-thia-l-azabicyclo[4.2.0]oct-
(3.0 per cent); 2-ene-2 -carboxylic add (decarbamoylcefoxitin),
2016 Cefpodoxime Proxetil 1-457
H COjH 0 CO,H
N/ T A '0 NHj
and epimer at C*
V n- V
-----^ J ★ ★
Cefpodoxime Proxetil ★ ★
i l T CHj (Ph. Eur. monograph 2341)
* * * * *
S ® and
anc epimer at C*
DEFINITION
COj H O
( 1R S)-1 -[ [( 1-Methylethoxy)carbonyl] oxy] ethyl (6R ,7R )-7-
H jC - 0 H n V | nh2 [[(22)-2-(2-aminothiazol-4-yI)-2-
(methoxyimino)acetyl]amino]-3-(methoxymethyl)-8-oxo-5-
( j I O^H S thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylate.
' — 0 ch3 Semi-synthetic product derived from a fermentation product.
Content
F. (6R,7S)-3-[(carbamoyloxy)methyl]-7-methoxy-7-[[(25)-2- 94.0 per cent to 102.0 per cent (anhydrous substance).
methoxy-2- (thiophen-2-yl) acetyl] amino] -8-oxo-5-thia-1-
azabicydo[4.2.0]oct-2-ene-2-carboxylic add CHARACTERS
((5)-methoxy cefoxitin), Appearance
White or pale yellow or light brown, amorphous powder.
Solubility
Very slightly soluble or practically insoluble in water, very
soluble in acetonitrile and in methanol, fredy soluble in
anhydrous ethanol.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison cefpodoxime proxetil CRS.
1-458 Cefpodoxkne Proxetil 2016
ch3 co2h
S ® and
an< epimer at C*
A
Y n ' V ' ' ' ° ' ch’
n^ a ^ n--J—¡\ j E. (1RS)-1 -[[(1 -methylethoxy)carbonyl]oxy]ethyl (6R,7R)-3-
H2N- f J T H HS (acetoxymethyl)-7-[[(2Z)-2~(2-aminothiazol-4-yl)-2-
(methoxyimino)acetyl] amino]-8-oxo-5-thia-1-
azabicydo[4.2.0]oct-2-ene-2-carboxy!ate (ACA-analogue of
A. (6R,7R)-7-[ [(2Z)-2-(2-aminothiazol-4-yl)-2- cefpodoxime proxetil),
(methoxyimino) acetyl] amino]-3-(methoxymethyI)-8-oxo-5-
thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic add
l‘,\ ,CH3
(cefpodoxime), k* V y °Y
C H, Y ° CH"
°y
Y °y
y CH3 N > -N 0 3
o. .0
CH, V ‘o ch3
and epimer at C*
and epimer at C*
G. (1RS)-1-[ [(1-methylethoxy) carbonyl] oxy] ethyl (6R,7R)-7-
[[(22)- [2-(2-acetylamino)thiazol-4-yI] -2-
(methoxyimino) acetyl] amino] -3- (methoxymethyl)-8-oxo-5-
C. (lÄS)-l-[[(l-methylethoxy)carbonyl]oxy]ethyl (6R,7R)-7- thia-l-azabicyclo [4.2.0] oct-2-ene-2 -carboxylate
[[(2Z)-2-(2-aminothiazol-4-yl)-2-
(N-acetyl-cefpodoxime proxetil),
(methoxyimino)acetyrjamino]-3-(methoxymethyl)-8-oxo-5-
thia- 1-azabicydo [4.2.0]oct-3-ene-2-carboxylate
(delta-2 -cefpodoxime proxetil),
1-460 Cefprozil Monohydrate 2016
TESTS
Related substances
Liquid chromatography (2.2.29). Prepare the solutions
immediately before use.
Test solution (a) Dissolve 0.125 g of the substance to be
examined in 1 mL of a 103 g/L solution of hydrochloric acid R
and dilute to 25.0 mL with mobile phase A.
Test solution (b) Dissolve 30.0 mg of the substance to be
examined in water R and dilute to 100.0 mL with the same
solvent.
Reference solution (a) Dilute 1.0 mL of test solution (a) to
100.0 mL with mobile phase A.
Reference solution (b) Dissolve 5 mg of cefprozüfor peak
identification CRS (containing impurities B, H and M) in
0.05 mL of a 103 g/L solution of hydrochloric acid R and add
and diastereoisomers at C1 and C1' 1 mL of mobile phase A.
Reference solution (c) Dissolve 3 mg of cefprozü CRS and 6 mg
H. mixture of the diastereoisomers of
of cefprozü impurity mixture CRS (containing impurities D
1 -[[(1 -methylethoxy)carbonyl]oxy]ethyl (6R,7R)-7-[[(2Z)-2-
and F) in 2 mL of a 103 g/L solution of hydrochloric acid R
[2-[[(2i?)-2-[[(2Z)-2-(2-aminothiazol-4-yl)-2-
and dilute to 50 mL with mobile phase A.
(methoxyimino)acetyl] amino] -2- [(2.R)-5-(methoxymethyl)-4-
[[1-[[(1 -methylethoxy) caibonyl] oxy] ethoxy] carbonyl] -3,6- Reference solution (d) Dissolve 30.0 mg of cefprozü CRS in
dihydro-2/f-1,3-thiazin-2-yl] acetyl] amino]thiazol-4-yl] -2- water R and dilute to 100.0 mL with the same solvent.
(methoxyimino)acetyI]amino]-3-(methoxymethyl)-8-oxo-5- Reference solution (e) Dissolve 10.0 mg of cefadroxü CRS
thia-1 -azabicyclo [4.2 .0] oct-2 -ene-2-carboxylate (impurity B) in water R and dilute to 20.0 mL with the same
(cefpodoxime proxetil dimer). solvent. Dilute 1.0 mL of the solution to 20.0 mL with
water R.
___________________________________________________________PhEur
Reference solution (f) Dissolve 10.0 mg of cefprozü
impurity A CRS in water R and dilute to 100.0 mL with the
same solvent. Dilute 1.0 mL of the solution to 10.0 mL with
water R.
Cefprozil Monohydrate ***** Column.
— size. I = 0.25 m, 0 = 4.6 mm;
(Ph Eur monograph 2342) *
— stationary phase, end-capped octadecylsüyl silica gelfor
chromatography R (5 (im);
— temperature: 40 °C.
Mobile phase:
— mobile phase A: dissolve 11.5 g of ammonium dihydrogen
phosphate R in water R, adjust to pH 4.4 with phosphoric
add R and dilute to 1000 mL with water R;
— mobile phase B: acetomtrüe R, mobile phase A (50:50 V!V)\
C is H ^ O jS , H20 407.4 121123-17-9
Time Mobile phase A Mobile phase B
Action and use (min) (per cent V/V) (per cent V7V)
Cephalosporin antibacterial 0 -8 81 19
8 - 20 81 -» 36 19->64
PhEur___________________________________________________________
20 -2 5 36 64
DEFINITION
Mixture of the 2 diastereoisomers of (6R,7R)-7-[[(2R)-2-
amino-2-(4-hydroxyphenyl) acetyl] amino] -8-oxo-3- [(1EZ) - Flow rate 1.0 mL/min.
prop- 1-enyl] -5-thia- 1-azabicydo [4.2.0] oct-2 -ene-2-carboxylic Detection Spectrophotometer at 230 nm.
add monohydrate. Injection 10 nL of test solution (a) and reference solutions (a),
Semi-synthetic product derived from a fermentation product. (b), (c), (e) and (f).
Content Identification of impurities Use the chromatogram supplied
96.0 per cent to 102.0 per cent (anhydrous substance). with cefprozUfor peak identification CRS and the
CHARACTERS chromatogram obtained with reference solution (b) to
identify the peaks due to impurities B} H and M; use the
Appearance
White or yellow, crystalline powder, slightly hygroscopic. chromatogram supplied with cefprozU impurity mixture CRS
and the chromatogram obtained with reference solution (c)
Solubility to identify the peaks due to impurities D and F; impurities G
Slightly soluble in water and in methanol, practically and I are identified by their rdative retention.
insoluble in acetone.
Relative retention With reference to cefprozil (Z)-isomer
IDENTIFICATION (retention time = about 7 min): impurity A = about 0.4;
Infrared absorption spectrophotometry (2.2.24). impurity B = about 0.5; impurity D = about 0.7;
Comparison cefprozU CRS. impurity F = about 0.9; cefprozil (£)-isomer = about 1.4;
2016 Cefprozil Monohydrate 1-461
impurity G = about 1.7; impurity H = about 2.0; Other detectable impurities (the fo llo w in g substances would, if
impurity I = about 2.1; impurity M = about 2.9. present at a sufficient level, be detected by one or other of
System suitability: reference solution (c): the tests in the monograph. They are limited by the general
— resolution: minimum 1.4 between die peaks due to acceptance criterion for other/unspecified impurities and/or
impurity F and cefprozil (2)-isomer. by the general monograph Substancesfor pharmaceutical use
Limits: (2034). It is therefore not necessary to identify these
— correction factor for the calculation of content, multiply the impurities for demonstration of compliance. See also 5.10.
peak area of impurity D by 2.3; Control of impurities in substancesfor pharmaceutical use): C, E,
— impurity B: not more than the area of the principal peak F , J , K ,L ,N .
in the chromatogram obtained with reference solution (e)
(0.5 per cent); H NHa
— impurities D, G, H , 1, M: for each impurity, not more
than 0.3 times the sum of the areas of the 2 principal
peaks in the chromatogram obtained with reference
H O ^
solution (a) (0.3 per cent);
— impurity A: not more than the area of the principal peak A. (2i?)-2-amino-2-(4-hydroxyphenyl)acetic add
in the chromatogram obtained with reference solution (f) (p-hydroxyphenylglycme),
(0.2 per cent);
— any other impurity, for each impurity, not more than
0.2 times the sum of the areas of the 2 principal peaks in
the chromatogram obtained with reference solution (a)
(0.2 per cent);
— total: maximum 2.0 per cent;
— disregard lirnic. 0.05 times the sum of the areas of the
2 principal peaks in the chromatogram obtained with
B. (6£,72?)-7-[[(22?)-2-ammo-2-
reference solution (a) (0.05 per cent). (4-hydroxyphenyl) acetyl] amino] -3-methyl-8-oxo-5-thia-1-
(E)-isomer ratio azabicydo [4.2.0] oct-2-ene-2-carboxylic add (cefadroxil),
Liquid chromatography (2.2.29) as described under Assay.
Determine the area of die peak due to the (£)-isomer in the
chromatogram obtained with test solution (b) and reference
solution (d). Calculate the ratio of the (£)-isomer to the sum
of both cefprozil isomers, as determined under Assay.
Limit:
— (E)-isomer ratio: 0.06 to 0.11.
Heavy metals (2.4.8) C. (6 RS)-3-(aminomethylene)-6 -
Maximum 20 ppm. (4-hydroxyphenyl)piperazine-2,5-dione,
1.0 g complies with test C. Prepare the reference solution COjH
using 2 m l, of lead standard solution (10 ppm Pb) R.
W ater (2.5.12)
3.5 per cent to 6.5 per cent, determined on 0.500 g.
V r
H 2N ” r K ^ C H j
H H
Sulfated ash (2.4.14)
Maximum 0.2 per cent, determined on 1.0 g.
D. (6i?,7R)-7-amino-8-oxo-3- [(1Z)-prop- 1-enyl]-5-thia-1-
ASSAY azabicydo[4.2.0]oct-2-ene-2-carboxylic add,
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modifications.
Mobile phase Mobile phase B, mobile phase A (18:82 V!V).
Detection Spectrophotometer at 280 nm.
Injection 10 |iL of test solution (b) and reference solution (d).
Run lime Twice the retention time of cefprozil (Z)-isomer.
Elution order (Z)-isomer, (£)-isomer. H2N H
co 2h
H nh2
h° Vï T ^
n ” H V C h3
H H
G. (2i?)-2-[(2i?)-2-amino-2-(4-hydroxyphenyl)acetyl]amino- M. (6i?,7i?)-7-[[(2i?)-2-amino-2-[4-
2-[(2i?)-4-carboxy-5- [( 12)-prop-1-enyl] -3j6-dihydro-2H - 1,3- [(ethoxycarbonyl)oxy] phenyl] acetyl] amino] -8 -oxo-3-[(12 )-
thiazin-2-yl]-acetic add. prop-1-enyl]-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic
add,
h nh2
co 2h
co 2h
h nh2
° ^ y " cH
H H
3
H H
h3c
H. (6i?j7i?)-7-[[(2i?)-2-[[(2Ä)-2-amino-2- N. (6Ä,7Ä)-7-[[(2JR)-2-amino-2-[4-
(4-hydroxyphenyl)acetyi] amino]-2- (ethoxycarbonyl)oxy] phenyl] acetyl] amino] -8-oxo-3- [(1E)-
(4-hydroxyphenyl) acetyl] amino]-8-oxo-3- [( 12)-prop-l -enyl]- prop-1-enyl]-5-thia-l -azabicydo [4.2.0] oct-2-ene-2-caiboxylic
5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic add. add.
PhEur
co 2h
.**★★
★
Cefradine ★ ★
(Ph. Eur. monograph 0814) *****
co 2h
I. (2i?)-2-[(2i?)-2-amino-2-(4-hydroxyphenyl)acetyl]amino-2-
[(2i?)-4-carboxy-5-[( 1£)-prop-l -enyl] -3j6-dihydro-2H-1,3- CHj
thiazin-2-yl]-acetic add,
Il H H
h nh2 O
co 2h
Compound Mol. Formula Mr
C15H19N3O4S 349.4
H H S
C1QH17N3O4S 347.4
J. (6£,72Q-7-[[(22Q-2-[[(2J$-2-amino-2-
(4-hydroxyphenyl) acetyl] amino] -2- ^16^21 N3O4S 351.4
(4-hydroxyphenyI) acetyl] amino] -8-oxo-3- [(1£)-prop-1-enyl] -
5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic add,
ASSAY
Liquid chromatography (2.2.29).
Test solution Dissolve 50.0 mg of die substance to be
examined in phosphate buffer solution p H 5.0 R and dilute to
100.0 mL with die same solution. B. (2i?)-amino(cyclohexa-l,4-dienyl)acetic add
Reference solution (a) Dissolve 50.0 mg of cefradine CRS (D-dihydrophenylglycme, cyclohexa-1,4-dienylglydne),
(containing 4 /,5'-dihydrocefradine) in phosphate buffer solution
p H 5.0 R and dilute to 100.0 mL with the same solution. COj H
Reference solution (b) Dissolve 5.0 mg of cefalexin
monohydrate CRS in phosphate buffer solution pH 5.0 R and HNH2H
dilute to 100.0 mL with the same solution.
Reference solution (c) Dilute 1 ml. of reference solution (a) to
10 m l. with phosphate buffer solution p H 5.0 R. Mix 5 mL of
this solution and 5 m l, of reference solution (b).
C. (6i?,7i?)-7-[[(2i?)-amino(cyclohexa-l,4-
Column:
dienyl) acetyl] amino] -3-methyl-8-oxo-5-thia-1-
— size: I = 0.10 m, 0 = 4.6 mm; azabicyclo[4.2.0]oct-2-ene-2-carboxylic add 5-oxide
— stationary phase: octadecylsHyl silica gelfor chromatography R (isomer 1),
(5 |im).
D. (6i?,7i?)-7-[[(2i?)-amino(cyclohexa-l,4-
Mobile phase methanol R, phosphate buffer solution p H 5.0 R
dienyl) acetyl] amino] -3-methyl-8-oxo-5-thia-1-
(25:75 V!V).
azabicyclo[4.2.0]oct-2-ene-2-carboxylic add 5-oxide
Flow rate 1.5 mL/min. (isomer 2 ),
Detection Spectrophotometer at 254 nm.
Injection 5 |xL. COj H
A. (6R,7R)-7-amino-3-methyl-8-oxo-5-thia-1-
azabicyclo [4.2.0] oct-2-ene-2-carboxylic add
(7-aminodeacetoxycephalosporanic add, 7-ADCA),
2016 Ceftazidime 1-465
Column'.
Ceftazidime Pentahydrate ★ ★ — sizer. I = 0.25 m, 0 = 4,6 mm;
Ceftazidime ***** — stationary phase, octadecylsüyl silica gelfor chromatography R
Reference solution (a) Dissolve 1.00 g of pyridine R in water R System suitability, reference
solution (b):
and dilute to 100.0 mL with the same solvent. Dilute 5.0 mL — resolution: minimum 1.5 between the peaks due to
of the solution to 200.0 mL with water R. Dilute 1.0 mL of impurity A and ceftazidime.
this solution to 100.0 mL with phosphate buffer solution. Calculate the content of ceftazidime (C22H 22N 60 7S2) taking
Reference solution (b) Dilute 1 mL of the test solution to into account the assigned content of C22H 22N 60 7S2 in
200 mL with phosphate buffer solution. To 1 mL of this ceftazidime CRS.
solution add 20 mL of reference solution (a) and dilute to STORAGE
200 mL with phosphate buffer solution.
In an airtight container. If the substance is sterile, store in a
Column: sterile, airtight, tamper-proof container.
— size: I = 0.25 m, 0 = 4.6 mm;
— stationary phase. octadecylsUyl silica gelfor chromatography R IMPURITIES
(5 nm). Specified impurities A,B, F, G
Mobile phase Mix 8 volumes of a 28.8 g/L solution of Other detectable impurities(the following substances would, if
ammonium dihydrogen phosphate R previously adjusted to present at a sufficient level, be detected by one or other of
pH 7.0 with ammonia R, 24 volumes of acetonitrüe R and the tests in the monograph. They are limited by the general
68 volumes of water R. acceptance criterion for other/unspecified impurities and/or
Flow rate 1.0 mL/min. by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these
Detection Spectrophotometer at 255 nm. impurities for demonstration of compliance. See also 5.10.
Injection 20 |iL. Control of impurities in substances for pharmaceutical use): C, E,
Run time 10 min. H.
System suitability, reference solution (b):
HjC\ CO,H
— resolution: m in im u m 7.0 between the peaks due to H3C-Y" h cor
ceftazidime and impurity F.
N
Limit.
— impurity F: not more than the area of the principal peak
in the chromatogram obtained with reference solution (a) H2N - f NY l f H H -
S"-^ 0 and epimer at C*
(500 ppm).
Heavy m etals (2.4.8) A. (2i?S,6i?,7i?)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-
Maximum 20 ppm. [(1 -carboxy-1 -methylethoxy)imino] acetyl] amino]-8-oxo-3-
1.0 g complies with test F. Prepare the reference solution [(pyridin-1-ium-1-yl) methyl] -5-thia-1-azabicyclo [4.2.0] oct-3-
using 2.0 ml, of lead standard solution (10 ppm Pb) R. ene-2 -carboxylate (A-2-ceftazidime),
W ater (2.5.12)
13.0 per cent to 15.0 per cent, determined on 0.100 g.
Bacterial endotoxins (2.6.14)
Less than 0.10 IU/mg, if intended for use in the manufacture
of parenteral preparations without a further appropriate
procedure for the removal of bacterial endotoxins.
ASSAY
Liquid chromatography (2.2.29). B. (6i?,7i?)-7-[[(2£)-2-(2-aminothiazol-4-yl)-2-[(l-carboxy- 1-
methylethoxy)imino] acetyl] amino]-8-oxo-3- [(pyridin- 1-him-
Test solution Dissolve 25.0 mg of the substance to be
1-yl) methyl]-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-
ex a m in ed in the mobile phase and dilute to 25.0 mL with
carboxylate,
the mobile phase.
Reference solution (a) Dissolve 25.0 mg of ceftazidime CRS in C02”
the mobile phase and dilute to 25.0 mL with the mobile
phase.
Reference solution (b) Dissolve 5.0 mg of ceftazidime for peak
H H
identification CR S (containing impurities A and G) in the
mobile phase and dilute to 5.0 mL with the mobile phase. C. (6i?,7i?)-7-amino-8-oxo-3-[(pyridin-l-him-l-yl)methyl]-5-
Column: thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylate,
— size. I = 0.15 m, 0 = 4.6 mm;
h3c ch3
— stationary phase: hexylsifyl silica gelfor chromatography R V
(5 ^m). o ch3
Mobile phase Dissolve 4.3 g of disodium hydrogen phosphate R
and 2.7 g of potassium dihydrogen phosphate R in 980 mL of
water R, then add 20 mL of acetonitrüe R.
Flow rate 2 mlVmin.
Detection Spectrophotometer at 245 nm.
Injection 20 |iL.
Run time 6 m in . E. (6Æ,7iQ-7-[[(22)-2-(2-aminothiazol-4-yl)-2-[[2-(l,l-
Relative retention With
reference to ceftazidime (retention dimethylethoxy)-l, 1-dimethyl-2 -
time = about 4.5 min): impurity A = about 0.7. oxoethoxy] imino] acetyl] amino] -8-oxo-3- [(pyridin-1-ium-1-
yl) methyl]-5-thia-1-azabicyclo [4.2.0] oct-2 -ene-2-carboxylate,
2016 Ceftazidime Pentahydrate with Sodium Carbonate for Injection 1-467
TESTS
Solution S
Dissolve 2.60 g in carbon dioxide-free water R and dilute to
F. pyridine. 20.0 mL with the same solvent.
Appearance of solution
H3C r n u Solution S is clear (2.2.1) and its absorbance (2.2.25) at
425 nm is not greater than 0.50.
„0
pH (2.2.3)
,CHO 5.0 to 7.5 for solution S.
H2N—^
Related substances
Liquid chromatography (2.2.29).
Test solution Suspend 0.150 g of the substance to be
G. 2-[ [[( 1Z )-1-(2-aminothiazol-4-yl)-2-[(oxoethyl) amino]-2 -
oxoethylidene] amino] oxy]-2-methyipropanoic acid, examined in 5 mL of acetonitrüe R, dissolve by adding
water R and dilute to 100 mL with water R.
och3 Reference solution (a) To 1.0 mL of the test solution add
5.0 mL of acetonitrüe R and dilute to 100.0 mL with water R.
Dilute 1.0 mL of this solution to 5.0 mL with water R.
Reference solution (b) In order to prepare impurity B in situ,
expose 5 mL of the test solution to ultraviolet light at
254 nm for about 24 h.
Reference solution (c) Suspend 3 mg of ceftazidime for peak
identification CRS (containing impurities A and G) in 0.5 ml.
H. (6i?,7i?)-7-[[(22)-2-(2-axninothiazol-4-yl)-2-[(2-met±ioxy- of acetonitrüe R, dissolve by adding water R and dilute to
1 j 1-dime thyl-2-oxoethoxy) imino] acetyl] amino] -8-oxo-3- 2 mL with water R.
[(pyridin-1-ium-1-yl) methyl]-5-thia-1-azabicyclo [4.2.0] oct-2- Column:
ene-2 -carboxylate. — sizer. I = 0.25 m, 0 = 4.6 mm;
PhEur — stationary phase: octadecylsQyl silica gelfor chromatography R
(5 pm);
— temperature: 40 °C.
Mobile phase:
— mobile phase A: solution con tainin g 3.6 g/L of disodium
Ceftazidime Pentahydrate with ;* * * *
hydrogen phosphate R and 1.4 g/L of potassium dihydrogen
Sodium Carbonate for Injection ***** phosphate R, adjusted to pH 3.4 with a 10 per cent VIV
(Ph Ettr monograph 2344) solution of phosphoric add 2?;
— mobile phase B: acetonitrüefor chromatography R;
Action and use
Cephalosporin antibacterial.
Time Mobile phase A Mobile phase B
Preparation
(min) (per cent V/V) (per cent V/V)
Ceftazidime Injection
0 -4 9 6 -»89 4 - » 11
Ceftazidime Eye Drops
4 -5 89 11
PhEur__________________________________________________________
5 -8 8 9 -»84 11-»16
DEFINITION
Sterile mixture of Ceftazidime pentahydrate (1405) and 8-11 8 4 -»80 16-»20
Anhydrous sodium carbonate (0773). 11-15 8 0 -»50 20 -»50
Semi-synthetic product derived from a fermentation product. 15-18 5 0 -»20 5 0 -»80
Content 18-22 20 80
— ceftazidime: 93.0 per cent to 105.0 per cent (dried and
carbonate-free substance);
— sodium carbonate: 8.0 per cent to 10.0 per cent. Flow rate 1.3 mlVmin.
CHARACTERS Detection Spectrophotometer at 254 nm.
Appearance Injection 10 jiL.
White or pale yellow powder. Relative retention With reference to ceftazidime (retention
Solubility. time = about 8 min): impurity F = about 0.4;
Freely soluble in water and in methanol, practically insoluble impurity G = about 0.8; impurity A = about 0.9;
in acetone. impurity B = about 1.4.
IDENTIFICATION Identification of impurities Use the chromatogram supplied
A. Examine the chromatograms obtained in the assay. with ceftazidime for peak identification CRS and the
chromatogram obtained with reference solution (c) to identify
Results The principal peak in the chromatogram obtained
the peaks due to impurities A and G; use the chromatogram
with the test solution is similar in retention time to the
obtained with reference solution (b) to identify the peak due
principal peak in the chromatogram obtained with reference
to impurity B.
solution (a).
B. It gives the reaction of carbonates (2.3.1).
1-468 Ceftazidime Pentahydrate with Sodium Carbonate for Injection 2016
— disregard limit. 0.25 times the area of the principal peak in (5 pm).
the chromatogram obtained with reference solution (a) Mobile phase Dissolve 4.3 g of disodium hydrogen phosphate R
(0.05 per cent); disregard the peak due to impurity F. and 2.7 g of potassium dihydrogen phosphate R in 980 mL of
water R, then add 20 mL of acetonitrile R.
Im purity F
Liquid chromatography (2.2.29). Prepare the solutions Flow rate 2 mL/min.
immediately before use. Detection Spectrophotometer at 245 nm.
Phosphate buffer solution Prepare a 10 per cent V/V solution of Injection 20 pL.
phosphate buffer solution p H 7.0 R4. Run time 6 min.
Test solution Dissolve 0.500 g of the substance to be Relative retention With reference to ceftazidime (retention
examined in phosphate buffer solution and dilute to time = about 4.5 min): impurity A = about 0.7.
100.0 mL with the same solution. System suitability: reference solution (b):
Reference solution (a) Dissolve 1.00 g of pyridine R in water R — resolution: m in im u m 1.5 between the peaks due to
and dilute to 100.0 mL with the same solvent. Dilute 5.0 mL impurity A and ceftazidime.
of the solution to 200.0 mL with water R. Dilute 1.0 mL of Calculate the content of ceftazidime (C22H 22N 60 7S2) taking
this solution to 100.0 mL with phosphate buffer solution. into account the assigned content of C 22H 22N 60 7S2 in
Reference solution (b) Dilute 1 .0 mL of the test solution to ceftazidime CRS.
200.0 mL with phosphate buffer solution. To 1.0 mL of this Sodium carbonate
solution add 20.0 mL of reference solution (a) and dilute to Atomic absorption spectrometry (2.2.23, Method I).
200.0 mL with phosphate buffer solution.
Caesium chloride buffer solution To 12.7 g of caesium chloride R
Column:
add 500 mL of water R and 86 mL of hydrochloric acid R and
— size. I = 0.25 m, 0 = 4.6 mm;
dilute to 1000.0 mL with water R.
— stationary phase: octadecylsilyl silica gel for chromatography R
Sodium standard solution (1000 mg/L) Dissolve 3.70 g of
(5 pm).
sodium nitrate R in water R and dilute to 500 mL with the
Mobile phase Mix 8 volumes of a 28.8 g/L solution of
same solvent, add 48.5 g of nitric acid R and dilute to
ammonium dihydrogen phosphate R previously adjusted to
1000 mL with water R.
pH 7.0 with ammonia R, 24 volumes of acetonitrUe R and
Test solution Dissolve 650.0 mg of the substance to be
68 volumes of water R.
exam in ed in water R and dilute to 100.0 mL with the same
Flow rate 1.0 mL/min. solvent. To 10.0 mL of this solution add 5.0 mL of caesium
Detection Spectrophotometer at 255 nm. chloride buffer solution and dilute to 50.0 mL with water R.
Injection 20 pL. Reference solution Into 4 identical flasks, each containing
Run time 10 min. 20.0 mT. of caesium chloride buffer solution, introduce
System suitability: reference solution (b): respectively 0 mL, 5.00 ml,, 10.00 mL and 15.00 mL of
— resolution: m in im u m 7.0 between the peaks due to sodium standard solution (1000 mg/L) and dilute to
ceftazidime and impurity F. 200.0 mL with water R.
Limit. Source Sodium hollow-cathode lamp.
— impurity F: not more than 6 rimes the area of the Wavelength 330.2 nm to 330.3 nm.
principal peak in the chromatogram obtained with Atomisation device Air-acetylene flame.
reference solution (a) (0.3 per cent).
Calculate the percentage content of sodium carbonate.
Loss on drying (2.2.32)
Maximum 13.5 per cent, determined on 0.300 g. Dry at STORAGE
25 °C at a pressure not ex cee d in g 0.67 kPa for 4 h then heat In a sterile, airtight, tamper-proof container, protected from
the residue at 100 °C at a pressure not exceeding 0.67 kPa light and humidity.
for 3 h. LABELLING
Bacterial endotoxins (2.6.14) The label states the percentage content m/m of ceftazidime.
Less than 0.10 IU/mg, if intended for use in the manufacture IMPURITIES
of parenteral preparations without a further appropriate Specified impurities A, B, F, G.
procedure for the removal of bacterial endotoxins.
2016 Ceftriaxone Sodium 1-469
H3C
Other detectable impurities (the following substances would, if h,c^ ' COîH
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by die general
acceptance criterion for other/unspecified impurities and/or ,CHO
by the general monograph Substancesfor pharmaceutical use h2n—^
(2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substancesfor pharmaceutical use): G. 2-[[[(12)-l-(2-aminothiazol-4-yl)-2- [(oxoethyl)amino]- 2-
C, E, H. oxoethylidene] amino] oxy] -2-methylpropanoic add,
h2n—^
and epimer at C*
Ni
carboxylate. vV - N SA Ni
h2n- ^ 11
31/j H20
DEFINITION
Disodium (6i?,7/Q-7-[[(22)-(2-aminothiazol-4-
yl) (methoxyimino) acetyl] amino] -3- [[(2-methyl-6-oxido-5-
oxo-2,5-dihydro-1,2,4-triazin-3-yl)sulfanyI] methyl] -8-oxo-5-
thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 3.5 hydrate.
Semi-synthetic product derived from a fermentation product
Content
E. (6R,7K)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2- [[2-( 1,1- 96.0 per cent to 102.0 per cent (anhydrous substance).
dimethylethoxy)-l, l-dimethyl-2 - CHARACTERS
oxoethoxy] imino] acetyl] amino] -8-oxo-3- [(pyridin-1-ium-1-
Appearance
yl)mediyl]-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylate,
Almost white or yellowish, slighdy hygroscopic, crystalline
powder.
Solubility
Freely soluble in water, sparingly soluble in methanol, very
slighdy soluble in anhydrous ethanol.
F. pyridine, IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24).
Comparison ceftriaxone sodium CRS.
1-470 Ceftriaxone Sodium 2016
O V fr J
^—J O and epimer at C*
solutions (a), (b) and (c).
Identification of impurities Use the chromatogram obtained
with reference solution (b) to identify the pair of peaks due
to impurity A and use the chromatogram obtained with
C20H22N4 0 1qS 510.5 64544-07-6 reference solution (c) to identify the pair of peaks due to
impurity B.
Action and use
Cephalosporin antibacterial. Relative retention With reference to cefuroxime axetil
diastereoisomer A: cefuroxime axetil
Preparations
diastereoisomer B = about 0.9, impurity A = about 1.2;
Cefuroxime Axetil Oral Suspension
impurity B = 1.7 and 2.1.
Cefuroxime Axetil Tablets System suitability: reference solution (b):
PhEur. — resolution: minimum 1.5 between the peaks due to
cefuroxime axetil diastereoisomer A and impurity A.
DEFINITION Limits:
Mixture of the 2 diastereoisomers of (li?5)-1-(acetyloxy)ethyl — impurity A: maximum 1.5 per cent for the sum of the pair
(6J?j7i?)-3-[(carbamoyloxy)methyi]-7-[[(Z)-2-(furan-2-yl)- of peaks;
2 (methoxyimino) acetyl] amino] -8-oxo-5-thia-1- — impurity B: maximum 1.0 per cent for the sum of the pair
azabicydo [4.2.0] oct-2-ene-2-carboxylate. of peaks;
Semi-synthetic product derived from a fermentation product. — impurity E: maximum 0.5 per cent;
Content — any other impurity: for each impurity, m axim um
96.0 per cent to 102.0 per cent (anhydrous substance). 0.5 per cent;
— total: maximum 3.0 per cent;
CHARACTERS — disregard limit: 0.05 times the area of the 2 principal peaks
Appearance in the chromatogram obtained with reference solution (a)
White or almost white powder. (0.05 per cent).
Solubility Diastereoisomer ratio
Slightly soluble in water, soluble in acetone, in ethyl acetate Liquid chromatography (2.2.29) as described in the test for
and in methanol, slightly soluble in ethanol (96 per cent). rdated substances.
IDENTIFICATION Limit Test solution:
A. Infrared absorption spectrophotometry (2.2.24). — the ratio of the area of the peak due to cefuroxime axetil
Comparison cefuroxime axetil CRS. diastereoisomer A to the sum of the areas of the peaks
B. Examine the chromatograms obtained in the assay. due to cefuroxime axetil diastereoisomers A and B is
between 0.48 and 0.55.
Results The principal peaks in the chromatogram obtained
with the test solution are similar in retention time and size to Acetone (2A. 24)
the peaks due to cefuroxime axetil diastereoisomers A and B M axim um 1.1 per cent
in the chromatogram obtained with reference solution (d). W ater (2.5.12)
TESTS M axim um 1.5 per cent, determined on 0.400 g.
Related substances ASSAY
Liquid chromatography (2.2.29): use the normalisation Liquid chromatography (2.2.29) as described in the test for
procedure. Prepare the test solution qnd reference solution (d) rdated substances with the following modifications.
immediately before use.
1-472 Cefuroxime Sodium 2016
ov
Injection Test solution and reference solution (d). ch3 y -o
System suitability Reference solution (d):
— resolution: minimum 1.5 between the peaks due to
cefuroxime axetil diastereoisomers A and B;
— repeatability: maximum relative standard deviation of
2.0 per cent for the sum of the peaks due to cefuroxime
axetil diastereoisomers A and B after 6 injections. E. (5ai?, 6 R)-6 - [[(2Z)-2-(furan-2-yl)-2-
Calculate the percentage content of C 20H 22N 4O 10S from the (methoxyimino)acetyl] amino] -5a, 6-dihydro-3H, 7H-azeto [2,1-
sum of the areas of the 2 diastereoisomer peaks and the ¿]furo[3,4-d] [1,3]thiazine-1,7(4/i)-dione
declared content of C20H 22N 4O 10S in cefuroxime axetil CRS. (descarbamoylcefuroxime laaone).
PhEur
STORAGE
In an airtight container, protected from light.
IMPURITIES
Specified impurities A, B, E. Cefuroxime Sodium ★ ★
★ ★
Other detectable impurities (the following substances would, if *****
(Ph. Eur. monograph 0992)
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by die general CH3 C02Na O
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use N "° nh2
(2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substancesfor pharmaceutical use): C, D.
C r Y iv Cefuroxime Injection
Cefuroxime Intracameral Injection
TESTS
Solution S
Dissolve 2.0 g in carbon dioxide-free water R and dilute to
C. R = CO-CCI3 : (6i?,7/?)-7-[[(Z)-2-(furan-2-yl)-2- 20.0 mL with the same solvent.
(methoxyimino) acetyl] amino]-8-oxo-3- Appearance of solution
[[[(trichloroacetyl) carbamoyl] oxy]methyl] -5-thia-1- Solution S is not more opalescent than reference
azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid, suspension II (2.2.1). The absorbance (2.2.25) of solution S
D. R = H: cefuroxime. measured at 450 nm is not greater than 0.25.
pH (2.2.3)
5.5 to 8.5.
Dilute 2 mT. of solution S to 20 mL with carbon dioxide-free
water R
2016 Cefuroxime Sodium 1-473
Specific optical rotation (2.2.7) Injection Testsolution (b) and reference solution (a).
+ 59 to + 66 (anhydrous substance). Calculate the percentage content of cefuroxime sodium.
Dissolve 0.500 g in acetate buffer solution p H 4.6 R and dilute STORAGE
to 25.0 mL with the same buffer solution.
In an airtight container. If the substance is sterile, store in a
Related substances sterile, airtight, tamper-proof container.
Liquid chromatography (2.2.29). Prepare the solutions
IMPURITIES
immediately before use or keep at 2-8 °C.
Test solution (a) Dissolve 25.0 mg of the substance to be
examined in water R and dilute to 25.0 ml, with the same
solvent.
Test solution (b) Dilute 5.0 mL of test solution (a) to
50.0 mL with water R.
Reference solution (a) Dissolve 25.0 mg of cefuroxime
sodium CRS in water R and dilute to 25.0 mL with the same
A. R = OH: (6J?,7/?)-7-[[(Z)-(furan-2-
solvent. Dilute 5.0 mL to 50.0 mL with water R. yl) (methoxyimino) acetyl] amino]-3-(hydroxymethyl)-8-oxo-5-
Reference solution (b) Place 20 mL of reference solution (a) in thia- 1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic add
a water-bath at 80 °C for 15 min. Cool and inject (descarbamoylcefuroxime),
immediately. B. R = 0-C 0-C H 3: (6.R,7i?)-3-[(acetyloxy)methyl]-7-[[(Z)-
Reference solution (c) Dilute 1.0 mL of test solution (a) to (furan-2-yl) (methoxyimino) acetyl] amino]-8-oxo-5-thia-1-
100.0 mL with water R. azabicydo[4.2.0]oct-2-ene-2-carboxylic add,
Column: C. R = H: (6£,7£)-7-[[(2Hfuran-2-
— size. I = 0.125 m, 0 = 4.6 mm; yl) (methoxyimino) acetyl] amino]-3-methyl-8-oxo-5-thia-1-
— stationary phase: hexylstlyl silica gelfor chromatography R azabicyclo [4.2.0] oct-2-ene-2-carboxylic add,
(5 nm). D. R = 0-C0-NH-C0-CCl3: (6£,7/?)-7-[[(2)-(furan-2-
Mobile phase Mix 1 volume of acetonitrile R and 99 volumes yl) (methoxyimino) acetyl] amino]-8-oxo-3-
of an acetate buffer solution pH 3.4, prepared by dissolving [[[(trichloroacetyl) carbamoyl] oxy] methyl] -5-thia-1-
6.01g of glacial acetic acid R and 0.68 g of sodium acetate R azabicyclo [4.2.0] oct-2-ene-2-carboxylic add,
in water R and diluting to 1000 mL with the same solvent.
Flow rate 1.5 mL/min.
Detection Spectrophotometer at 273 nm.
Injection 20 jiL loop injector; inject test solution (a) and
reference solutions (b) and (c).
Run time 4 times the retention time of cefuroxime.
System suitability: reference solution (b):
— resolution: minimum 2.0 between the peaks due to E. R = 0-C 0-N H 2: (6R,7K)-3- [(carbamoyloxy)methyl]-7-
cefuroxime and impurity A. [ [(E)-(furan-2-yl) (methoxyimino) acetyl] amino] -8-oxo-5-thia-
Limits:
1-azabicydo [4.2.0] oct-2-ene-2-carboxylic add (trans-
— impurity A: not more than the area of the principal peak cefuroxime),
in the chromatogram obtained with reference solution (c) F. R = OH: (6fl,7i?)-7-[[(£)-(furan-2-
( 1.0 per cent); yl) (methoxyimino) acetyl] amino] -3-(hydroxymethyl)-8-oxo-5-
— any other impurity, not more than the area of the principal thia- 1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic add,
peak in the chromatogram obtained with reference G. R = O-CO-CH3: (6i?,7i?)-3-[(acetyloxy)methyl]-7-[[(£)-
solution (c) ( 1.0 per cent); (furan-2-yI) (methoxyimino) acetyl] amino]-8-oxo-5-thia-1-
— total: not more than 3 times the area of the principal peak azabicyclo[4.2.0]oct-2-ene-2-carboxylic add,
in the chromatogram obtained with reference solution (c)
(3.0 per cent);
— disregard limit. 0.05 times the area of the principal peak in
the chromatogram obtained with reference solution (c)
(0.05 per cent).
Af-THmethylanfline (2.4.26, Method E)
Maximum 20 ppm.
2-Ethylhexanoic acid (2.4.28)
Maximum 0.5 per cent mim. H. (5ai?,6i?)-6-[[CZ)-(furan-2-
yl) (methoxyimino)acetyl] amino] -5 a, 6-dihydro-3/f,7//-
W ater (2.5.12) azeto [2,1-6]furo [3,4-d\ [1,3] thiazine-1,7(4i/)-di°ne,
Maximum 3.5 per cent, determined on 0.400 g.
Bacterial endotoxins (2.6.14)
Less than 0.10 IU/mg, if intended for use in the manufacture
of parenteral preparations without a further appropriate
procedure for the removal of bacterial endotoxins.
ASSAY
Liquid chromatography (2.2.29) as described in the test for I. (Z)-(furan-2 -yl) (methoxyimino) acetic add.
related substances with the following modification. __________________________________________________________ PhEur
1-474 Celecoxib 2016
PhEur__________________________
Urne Mobik phase A Mobile phase B
DEFINITION (min) (per cent V/V) (per cent V/V)
3-[3-Acetyl-4- [(2i?5)-3- [( 1, l-dimethylethyl)amino]-2- 0 -5 0 100-»80 0 + 20
hydroxypropoxy]phenyI]-l, 1-diethylurea hydrochloride. 50-51 80-» 100 20->0
Content 51 -65 100 0
99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS Flow rate 1.4 mUmin.
Appearance
Detection Spectrophotometer at 232 nm.
White or very slightly yellow, crystalline powder.
Injection 10 (iL.
Solubility
Identification of impurities Use the chromatogram supplied
Freely soluble in water and in methanol, soluble in ethanol
(96 per cent), very slightly soluble in methylene chloride. with celiprololfor peak identification CRS and the
chromatogram obtained with reference solution (d) to
It shows polymorphism (5.9). identify the peaks due to impurities B, E and F.
IDENTIFICATION Relative retention With reference to celiprolol (retention
A. Infrared absorption spectrophotometry (2.2.24). time = about 10 min): impurity A = about 0.3;
Comparison celiprolol hydrochloride CRS. impurity D = about 0.7; impurity G = about 1.2 ;
1-476 Cellacefate 2016
H3CJ H
1 mL of 0.1 M sodium hydroxide is equivalent to 41.60 mg of
C 20H 34CIN3O4.
STORAGE G. 3- [3-acetyI-4- [[(i?5) -oxiranyl] methoxy] phenyl] -1,1-
Protected from light. diethylurea.
IM PURITIES PftEur
Specified impuritier. A, B, C, D, E, F, G, H, I.
Cellacefate *****
and enantiomer ★ ★
R1. * * * * *
(Cellulose Acetate Phthalate, Ph Eitr monograph 0314)
9004-38-0
A. R1 = H, R2 = NH-C(CH3)3:
Action and use
l-[5-amino-2-[(2.&S)-3-[(l, l-dimethylethyl)amino]-2-
Enteric coating in pharmaceutical products.
hydroxypropoxy]phenyl] ethanone,
C. R 1 = CO-NH-C(CH3)3, R2 = NH-C(CH3)3: PtiEur.
1-[3-acetyl-4-[(2i?5)-3-[(l, 1-dimethylethyl)amino] -2- DEFINITION
hydroxypropoxy]phenyl]-3-(l,l-dimethylethyl)urea, Partly O-acetylated and Ophthalylated cellulose.
D. R1 = CO-N(C 2H 5)2, R2 = N(C 2H 5)2: 3-[3-acetyl-4-
Content
[(2J?5)-3-(diethylamino)-2-hydroxypropoxy]phenyl]-l, 1-
diethylurea,
— phthaloyl groups (CgHsOj; M r 149.1): 30.0 per cent to
36.0 per cent (anhydrous and acid-free substance);
H. R 1 = CO-N(C 2H 5)2, R2 = Bn 3-[3-acetyl-4-[(2.RS)-3- — acetyl groups (C2H 3 0 ; M T 43.04): 21.5 per cent to
bromo-2 -hydroxypropoxy]phenyl]-l, 1-diethylurea 26.0 per cent (anhydrous and add-free substance).
(bromhydrin compound),
2016 Cellacefate 1-477
Loss on drying
Dispersible Cellulose When dried to constant weight at 105°, loses not more than
Action and use 8.0% of its weight. Use 1 g.
Pharmaceutical excipient. Sulfated ash
Not more than 5.0%, Appendix IX A. Use 2 g.
DEFINITION
ASSAY
Dispersible Cellulose is a colloid-forming, attrited mixture of
Microcrystalline Cellulose and Carmellose Sodium. Heat 2 g with 75 mL of anhydrous acetic add under a reflux
condenser for 2 hours, cool and carry out Method I for rum-
Content o f carmellose sodium aqueous titration, Appendix V1H A, determining the end point
75.0 to 125.0% w/w of the stated amount. potentiometrically. Each mL of 0.1m perchloric add FS is
CHARACTERISTICS equivalent to 29.6 mg of carmellose sodium.
A white or off-white, coarse or fine powder. STORAGE
Disperses in water producing a white, opaque dispersion or Dispersible Cellulose should be stored at a temperature of 8 °
gel; practically insoluble in organic solvents and in dilute to 15°.
acids.
LABELLING
IDENTIFICATION The label states (1) the percentage w/w of Carmellose
A. Mix 6 g with 300 mL of water stirring at Sodium; (2) the viscosity of a dispersion in water of a stated
18.000 revolutions per minute for 5 m in u tes. A white, percentage w/w of Carmellose Sodium.
opaque dispersion is obtained which does not produce a
supernatant liquid.
B. Add several drops of the dispersion obtained in test A to a
10% w/v solution of aluminium chloride. Each drop forms a
white, opaque globule which does not disperse on standing.
Microcrystalline Cellulose *****
(Ph. Eur. monograph 0316)
*+ +**
C. Add 2 mL of iodinated potassium iodide solution to the
dispersion obtained in test A. No blue or purplish colour is
produced.
D. The solution obtained in the test for Heavy metals yields
the reactions characteristic of sodium salts, Appendix VI,
except that in test A the white precipitate produced may not
be dense.
TESTS
Acidity or alkalinity
pH of the dispersion obtained in the test for Apparent
viscosity, 6.0 to 8.0, Appendix V L.
C6«H10n+2O5n+l 9004-34-6
Solubility
Add 50 mg to 10 mL of ammoniacal solution of copper Action and use
tetrammine and shake. It dissolves completely leaving no Excipient.
residue.
PhEur___________________________________________________________
A pparent viscosity
60 to 140% of the declared value when determined by the DEFINITION
following method. Calculate the quantity (x g) needed to Purified, partly depolymerised cellulose prepared by treating
prepare exactly 600 g of a dispersion of the stated percentage alpha-cellulose, obtained as a pulp from fibrous plant
w/w, with reference to the dried substance. To (600-*) g of material, with mineral acids.
water at 23° to 25° contained in a 1000 mL high-speed
CHARACTERS
blender bowl add x g of the substance b ein g examined,
Appearance
stirring at reduced speed, taking care to avoid contacting the
White or almost white, fine or granular powder.
sides of the bowl with the powder. C on tin u e stirring at low
speed for 15 seconds after the addition and then stir at Solubility
18,000 revolutions per minute for exactly 2 minutes. Practically insoluble in water, in acetone, in anhydrous
Immerse the appropriate spindle of a rotational viscometer, ethanol, in toluene, in dilute acids and in a 50 g/L solution of
switch on after 30 seconds and after a further 30 seconds sodium hydroxide.
determine the viscosity, Appendix V H, Method IE, using a IDENTIFICATION
speed of 20 revolutions per m in u te (2.09 radians per A. Place about 10 mg on a watch-glass and disperse in 2 mL
second). of iodinated zinc chloride solution R. The substance becomes
Heavy metals violet-blue.
To the residue obtained in the test for Sulfated ash add B. The degree of polymerisation is not more than 350.
1 mL of hydrochloric add, evaporate to dryness on a water Transfer 1.300 g to a 125 mL conical flask. Add 25.0 mL of
bath and dissolve the residue in 20 mL of water. 12 mL of water R and 25.0 mT, of cupriethylenediamine hydroxide
the resulting solution complies with limit test A for heavy solution R. Immediately purge the solution with nitrogen R,
metals, Appendix VII. Use lead standard solution (1 ppm) Pb to insert the stopper and shake until completely dissolved.
prepare the standard (10 ppm). Transfer an appropriate volume of the solution to a suitable
capillary viscometer (2.2.9). Equilibrate the solution at
25 ± 0.1 °C for at least 5 min. 'Record the flow time fa) in
2016 Cellulose 1-479
1.1 0.098 0.106 0.115 0.125 0.134 0.143 0.152 0.161 0.170 0.180
lO 0.189 0.198 0.207 0J16 0025 0.233 0042 0050 0059 0068
U 0076 0.285 0-293 0302 0310 0318 0326 0334 0342 0350
1.4 0358 0.367 0375 0383 0391 0399 0.407 0.414 0.422 0.430
13 0.437 0.445 0.453 0.460 0.465 0.476 0.484 0.491 0.499 0307
1.6 0.515 0322 0329 0336 0344 0351 0358 0366 0373 0380
1.7 0387 0395 0.602 0.608 0.615 0.622 0.629 0.636 0.642 0349
1.8 0.656 0.663 0.670 0.677 0.683 0.690 0.697 0.704 0.710 0.717
1.9 0.723 0.730 0.736 0.743 0.749 0.756 0.762 0.769 0.775 0.7X2
2.0 0.788 0.795 0.802 0.809 0.815 0.821 0.827 0.833 0.840 0346
2.1 0.852 0.858 0.864 0370 0376 0.882 0.888 0.894 0.900 0306
20 0.912 0.918 0.924 0.929 0.935 0.941 0.948 0.953 0359 0365
23 0.971 0.976 0.983 0.988 0394 1.000 1.006 1.011 1.017 1.022
2.4 1.028 1.033 1.039 1.044 1.050 1.056 1.061 1.067 1.072 1378
23 1.083 1.089 1.094 1.100 1.105 1.111 1.116 1.121 1.126 1.131
2.6 1.137 1.142 1.147 1.153 1.158 1.163 1.169 1.174 1.179 1.184
2.7 1.190 1.195 1-200 1305 1010 1.215 1020 1025 1030 1035
2.8 1040 1.245 1050 1-255 1060 1065 1070 1075 1080 1085
IS 1090 1.295 1300 1305 1310 1314 1319 1324 1329 1333
3.0 1-338 1-343 1348 1352 1357 1362 1367 1371 1376 1381
3.1 1386 1.390 1395 1.400 1.405 1.409 1.414 1.418 L423 1.427
30 1.432 1.436 1.441 1.446 1.450 1.455 1.459 1.464 1.468 1.473
33 1.477 1.482 1.486 1.491 1.496 1300 1304 1308 1313 1317
3.4 1.521 1325 1329 1333 1337 1342 1346 1350 1354 1358
33 1362 1366 1370 1375 1379 1383 1387 1391 1395 L600
3.6 1.604 1.608 1.612 1.617 1.621 1.625 1.629 1.633 1-637 1.642
3.7 1.646 1.650 1-654 1.658 1.662 1.666 1.671 1.675 1.679 1383
3.8 L687 1.691 1.695 1.700 1.704 1.708 1.712 1.715 1.719 1.723
33 1.727 1.731 1.735 1.739 1.742 1.746 L750 1.754 1.758 1.762
4.0 1.765 1.769 1.773 1.777 1.781 1.785 1.789 1.792 1.796 1300
4.1 1.804 1.808 1.811 1315 1.819 1.822 1.826 1.830 1.833 1337
40 1.841 1.845 1348 1352 1.856 1359 1.863 L867 1370 1374
4J 1378 1.882 1385 1389 1.893 1396 1300 L904 1307 1311
4.4 1.914 L918 1321 1325 1329 1332 1336 1339 1343 1346
43 1.950 1.954 1.957 1.961 1364 1.968 1.971 1.975 1379 1382
4.6 1.986 1.989 1393 1396 2.000 2.003 2.007 1010 2X13 2X17
4.7 2.020 2.023 2.027 2.030 2.033 2.037 2.040 2.043 2.047 2.050
4.8 2.053 2.057 2.060 2.063 2.067 2.070 2.073 2.077 2.080 2.083
4.9 2.087 2.090 2X93 2.097 2.100 2.103 2.107 1110 2.113 2.116
5.0 2.119 2.122 2.125 2.129 2.132 2.135 2.139 2.142 2.145 2.148
5.1 2.151 2.154 2.158 2.160 2.164 2.167 2.170 2.173 2.176 2.180
SO 2.183 2.186 2.190 2.192 2.195 2.197 2000 2003 2006 2009
53 2-212 2015 2-218 2021 2024 2.227 2030 2033 2036 2040
5.4 2-243 1246 2-249 2052 2055 2.258 2061 2064 2067 2070
53 2073 2-276 2079 2082 2085 2088 2091 2094 2097 2300
5.6 2.303 2306 2309 2312 2315 2318 2320 2324 2326 2329
5.7 2-332 2335 ¿338 2-341 2344 2347 2350 2353 2355 2358
53 2361 2364 2367 2370 2373 2376 2379 2.382 2384 2387
5.9 2.390 2393 2396 2.400 2.403 2.405 2.408 2.411 2.414 2.417
6.0 2.419 2.422 2.425 2.428 2.431 2.433 2.436 2.439 2.442 2.444
6.1 2.447 2.450 2.453 145« 2.458 2.461 1464 2.467 2.470 2.472
60 2.475 2.478 2.481 2.483 2.486 2.489 2.492 2.494 2.497 2300
63 2-503 2305 2308 2311 2313 2316 2318 2321 2324 2326
6.4 2329 2332 2334 2337 2340 2342 2345 2347 2.550 2353
6.5 2355 2.558 2361 2363 2366 2368 2371 2374 2376 2379
6.6 2381 2384 2387 2390 2392 ‘ 2.595 2397 2.600 2.603 2.605
6.7 2.608 2.610 2.613 2.615 2.618 2.620 2.623 2.625 2.627 2.630
6.8 2.633 2.635 2.637 2.640 2.643 2.645 2.648 2.650 2.653 2.655
6.9 2.658 2.660 ' 2.663 2.665 2.668 2.670 2.673 2.675' 2378 2380
1-480 Cellulose 2016
M.
n* 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
7.0 2.683 2.685 2.687 2.690 2.693 2.695 2.698 2.700 2.702 2.705
7.1 2.707 2.710 2.712 2.714 2.717 2.719 2.721 2.724 2.726 2.729
72 2.731 2.733 2.736 2.738 2.740 2.743 2.745 2.748 2.750 2.752
7.3 2.755 2.757 2.760 2.762 2.764 2.767 2.769 2.771 2.774 2.776
7.4 2.779 2.781 2.783 2.786 2.788 2.790 2.793 2.795 2.798 2.800
73 2.802 2.805 2.807 2.809 2.812 2.814 2.816 2.819 2.821 2.823
7.6 2.826 2.828 2.830 2.833 2.835 2.837 2.840 2.842 2.844 2.847
7.7 2.849 2.851 2.854 2.856 2.858 2.860 2.863 2.865 2.868 2.870
7.8 2.873 2.875 2.877 2.879 2.881 2.884 2.887 2.889 2.891 2.893
7.9 2.895 2.898 2.900 2.902 2.905 2.907 2.909 2.911 2.913 2.915
8.0 2.918 2.920 2.922 2.924 2.926 2.928 2.931 2.933 2.935 2.937
8.1 2.939 2.942 2.944 2.946 2.948 2.950 2.952 2.955 2.957 2.959
8.2 2.961 2.963 2.966 2.968 2.970 2.972 2.974 2.976 2.979 2.981
8.3 2.983 2.985 2.987 2.990 2.992 2.994 2.996 2.998 3.000 3.002
8.4 3.004 3.006 3.008 3.010 3.012 3.015 3.017 3.019 3.021 3.023
8.5 3.025 3.027 3.029 3.031 3.033 3.035 3.037 3.040 3.042 3.044
8.6 3.046 3.048 3.050 3.052 3.054 3.056 3.058 3.060 3.062 3.064
8.7 3.067 3.069 3.071 3.073 3.075 3.077 3.079 3.081 3.083 3.085
8.8 3.087 3.089 3.092 3.094 3.096 3.098 3.100 3.102 3.104 3.106
8.9 3.108 3.110 3.112 3.114 3.116 3.118 3.120 3.122 3.124 3.126
9.0 3.128 3.130 3.132 3.134 3.136 3.138 3.140 3.142 3.144 3.146
9.1 3.148 3.150 3.152 3.154 3.156 3.158 3.160 3.162 3.164 3.166
92 3.168 3.170 3.172 3.174 3.176 3.178 3.180 3.182 3.184 3.186
93 3.188 3.190 3.192 3.194 3.196 3.198 3.200 3.202 3-204 3.206
9.4 3.208 3.210 3-212 3.214 3-215 3.217 3-219 3-221 3.223 3.225
9.5 3.227 3229 3-231 3.233 3.235 3.237 3J239 3.241 3-242 3.244
9.6 3.246 3.248 3.250 3252 3.254 3.256 3258 3260 3.262 3.264
9.7 3.266 3.268 3-269 3211 3273 3275 3277 3279 3.281 3-283
9.8 3.285 3.287 3-289 3-291 3.293 3295 3297 3298 3-300 3-302
9.9 3.304 3.305 3307 3.309 3.311 3.313 3.316 3.318 3.320 3-321
seconds between the 2 marks on the viscometer. Calculate Determine the relative viscosity (rjrJ) of die substance to be
the kinematic viscosity (vx) of the solution using the following examined using the following expression:
expression:
vi/v?
t\ (kx)
¿i is the viscometer constant.
Determine the intrinsic viscosity (faJJ by interpolation, using
Dilute a suitable volume of cupriethylenediamine hydroxide the intrinsic viscosity table (Table 0316.-1).
solution R with an equal volume of water R and measure the
flow time fe) using a suitable capillary viscometer. Calculate Calculate the degree of polymerisation (P) using the
the kinematic viscosity (V2) of the solvent using the following following expression:
expression:
95 [t?]c
ta (fca) m [(100 - 6 ) /100]
M,
*1* 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
10 332 3.34 3.36 3.37 3.39 3.41 3.43 3.45 3.46 3.48
11 330 3.52 3.53 3.55 3.56 3.58 3.60 3.61 3.63 3.64
12 3.66 3.68 3.69 3.71 3.72 3.74 3.76 3.77 3.79 3.80
13 3.80 3.83 3.85 3.86 3.88 3.89 3.90 3.92 3.93 3.95
14 3.96 3.97 3.99 400 4.02 403 404 406 4.07 4.09
15 4.10 4.11 4.13 414 4.15 417 418 4.19 4.20 4.22
16 423 4-24 425 427 4.28 429 4.30 4.31 4.33 4.34
17 4.35 4.36 4.37 4.38 439 441 442 443 444 4.45
18 4.46 4.47 448 449 4.50 4.52 4.53 4.54 4.55 456
19 4.57 458 459 460 4.61 4.62 4.63 4.64 4.65 466
M.
1m 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
1.1 0.098 0.106 0.115 0.125 0.134 0.143 0.152 0.161 0.170 0.180
1.2 0.189 0.198 0.207 0.216 0.225 0.233 0.242 0.250 0-259 0.268
1.3 0.276 0.285 0.293 0302 0310 0.318 0326 0.334 0342 0350
1.4 0.358 0.367 0.375 0383 0391 0.399 0.407 0.414 0.422 0.430
1.5 0.437 0.445 0.453 0.460 0.468 0.476 0.484 0.491 0.499 0.507
1.6 0.515 0.522 0.529 0.536 0344 0.551 0.558 0.566 0373 0380
1.7 0.587 0.595 0.602 0.608 0.615 0.622 0.629 0.636 0.642 0.649
1.8 0.656 0.663 0.670 0.677 0.683 0.690 0.697 0.704 0.710 0.717
1.9 0.723 0.730 0.736 0.743 0.749 0.756 0.762 0.769 0.775 0.782
2.0 0.788 0.795 0.802 0.809 0.815 0.821 0.827 0.833 0.840 0.846
2.1 0.852 0.858 0.864 0.870 0.876 0.882 0.888 0.894 0.900 0.906
2.2 0.912 0.918 0.924 0.929 0.935 0.941 0.948 0.953 0.959 0.965
2.3 0.971 0.976 0.983 0.988 0.994 1.000 1.006 1.011 1.017 1.022
2.4 1.028 1.033 1.039 1.044 1.050 1.056 1.061 1.067 1.072 1.078
2.5 1.083 1.089 1.094 1.100 1.105 1.111 1.116 1.121 1.126 1.131
2.6 1.137 1.142 1.147 1.153 1.158 1.163 1.169 1.174 1.179 1.184
2.7 1.190 1.195 1.200 1.205 U 10 1.215 1.220 1-225 1-230 1.235
2.8 1.240 1.245 1.250 1.255 1.260 1.265 1.270 1.275 1.280 1-285
2.9 1.290 1.295 1300 1305 1310 1.314 1319 1324 1329 1.333
3.0 1.338 1.343 1.348 1352 1357 1.362 1.367 1.371 1376 1381
3.1 1386 1390 1395 1.400 1.405 1.409 1.414 1.418 1.423 1.427
3.2 1.432 1.436 1.441 1.446 1.450 1.455 1.459 1.464 1.468 1.473
3-3 1.477 1.482 1.486 1.491 1.496 1.500 1.504 1308 1313 1317
3.4 1321 1.525 1.529 1333 1337 1.542 1.546 1.550 1354 1358
3.5 1.562 1.566 1.570 1375 1.579 1.583 1.587 1.591 1395 1.600
3.6 1.604 1.608 1.612 1.617 1.621 1.625 1.629 1.633 1.637 1.642
3.7 1.646 1.650 1.654 1.658 1.662 1.666 1.671 1.675 1.679 1.683
3.8 1.687 1.691 1.695 1.700 1.704 1.708 1.712 1.715 1.719 1.723
3.9 1.727 1.731 1.735 1.739 1.742 1.746 1.750 1.754 1.758 1.762
seconds between the 2 marks on the viscometer. Calculate where k2 is the viscometer constant.
the kinematic viscosity (vj) of the solution using the following Determine the relative viscosity of the substance to be
expression: examined using the following expression:
t \ (k i ) V\/V2
where kx is the viscometer constant. Determine the intrinsic viscosity by interpolation, using
Dilute a suitable volume of citprietkylenediamine hydroxide the intrinsic viscosity table (Table 0315.-1).
solution R with an equal volume of water R and measure the Calculate the degree of polymerisation (P) vising the
flow time (¿2) using a suitable capillary viscometer. Calculate following expression:
the kinematic viscosity (va) of the solvent using the following
expression: 95 [ij]c
m [(100 - b) /100]
£2 (fea)
1-484 Cellulose 2016
M.
0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
n*
40 1.765 1.769 1.773 1.777 1.781 1.785 1.789 1.792 1.796 1.800
4.1 1.804 1.808 1.811 1.815 1.819 1.822 1.826 1.830 1.833 1.837
42 1.841 1.845 1.848 1.852 1.856 1.859 1.863 1.867 1.870 1.874
43 1.878 1.882 1.885 1.889 1.893 1.896 1.900 1.904 1.907 1.911
4.4 1.914 1.918 1.921 1.925 1.929 1.932 1.936 1.939 1.943 1.946
4.5 1.950 1.954 1.957 1.961 1.964 1.968 1.971 1.975 1.979 1.982
4.6 1.986 1.989 1.993 1.996 2.000 2.003 2.007 2.010 2.013 2.017
4.7 2.020 2.023 2.027 2.030 2.033 2.037 2.040 2.043 2.047 2.050
48 2.053 2.057 2.060 2.063 2.067 2.070 2.073 2.077 2.080 2.083
4.9 2.087 2.090 2.093 2.097 2.100 2.103 2.107 2.110 2.113 2.116
5.0 2.119 2.122 2.125 2.129 2.132 2.135 2.139 2.142 2.145 2.148
5.1 2.151 2.154 2.158 2.160 2.164 2.167 2.170 2.173 2.176 2.180
5.2 2.183 2.186 2.190 2.192 2.195 2.197 2.200 2.203 2.206 2.209
5.3 2.212 2J215 2.218 2.221 2^24 2.227 2.230 2.233 2236 2.240
5.4 2.243 2-246 2.249 2252 2.255 2.258 2.261 2.264 2.267 2.270
5.5 ’ 2.273 2.276 2279 2282 2285 2.288 2.291 2.294 2297 2.300
5.6 2.303 2.306 2.309 2.312 2.315 2.318 2.320 2.324 2.326 2.329
5.7 2.332 2.335 2.338 2.341 2.344 2.347 2.350 2.353 2.355 2.358
5.8 2.361 2.364 2.367 2.370 2.373 2.376 2.379 2.382 2.384 2.387
5.9 2.390 2.393 2.396 2.400 2.403 2.405 2.408 2.411 2.414 2.417
6.0 2.419 2.422 2.425 2.428 2.431 2.433 2.436 2.439 2.442 2.444
6.1 2.447 2.450 2.453 2.456 2.458 2.461 2.464 2.467 2.470 2.472
62 2.475 2.478 2.481 2.483 2.486 2.489 2.492 2.494 2.497 2.500
6.3 2.503 2.505 2308 2.511 2.513 2.516 2.518 2.521 2.524 2.526
6.4 2.529 2.532 2.534 2.537 2.540 2.542 2.545 2.547 2.550 2.553
6.5 2.555 2.558 2.561 2.563 2.566 2.568 2.571 2.574 2.576 2.579
6.6 2.581 2.584 2.587 2.590 2.592 2.595 2.597 2.600 2.603 2.605
6.7 2.608 2.610 2.613 2.615 2.618 2.620 2.623 2.625 2.627 2.630
6.8 2.633 2.635 2.637 2.640 2.643 2.645 2.648 2.650 2.653 2.655
6.9 2.658 2.660 2.663 2.665 2.668 2.670 2.673 2.675 2.678 2.680
7.0 2.683 2.685 2.687 2.690 2.693 2.695 2.698 2.700 2.702 2.705
7.1 2.707 2.710 2.712 2.714 2.717 2.719 2.721 2.724 2.726 2.729
72 2.731 2.733 2.736 2.738 2.740 2.743 2.745 2.748 2.750 2.752
7.3 2.755 2.757 2.760 2.762 2.764 2.767 1769 2.771 2.774 2.776
7.4 2.779 2.781 2.783 2.786 2.788 2.790 2.793 2.795 2.798 2.800
7.5 2.802 2.805 2.807 2.809 2.812 2.814 2.816 2.819 2.821 2.823
7.6 2.826 2.828 2.830 2.833 2.835 2.837 2.840 2.842 2.844 2.847
7.7 2.849 2.851 2.854 2.856 2.858 2.860 2.863 2.865 2.868 2.870
7.8 2.873 2.875 2.877 2.879 2.881 2.884 2.887 2.889 2.891 2.893
7.9 2.895 2.898 2.900 2.902 2.905 2.907 2.909 2.911 2.913 2.915
2016 Cellulose 1-485
w,
1* 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
8.0 2.918 2.920 2.922 1924 1926 1928 1931 1933 2.935 1937
8.1 2.939 2.942 2.944 2.946 1948 2.950 1952 1955 2.957 2.959
8.2 2.961 2.963 2.966 1968 1970 2.972 1974 1976 2.979 2.981
8.3 2.983 2.985 2.987 1990 1992 2.994 1996 1998 3.000 3.002
8.4 3.004 3.006 3.008 3.010 3.012 3.015 3.017 3.019 3.021 3.023
8.5 3.025 3.027 3.029 3.031 3.033 3.035 3.037 3.040 3.042 3.044
8.6 3.046 3.048 3.050 3.052 3.054 3.056 3.058 3.060 3.062 3.064
8.7 3.067 3.069 3.071 3.073 3.075 3.077 3.079 3.081 3.083 3.085
8.8 3.087 3.089 3.092 3.094 3.096 3.098 3.100 3.102 3.104 3.106
8.9 3.108 3.110 3.112 3.114 3.116 3.118 3.120 3.122 3.124 3.126
9.0 3.128 3.130 3.132 3.134 3.136 3.138 3.140 3.142 3.144 3.146
9.1 3.148 3.150 3.152 3.154 3.156 3.158 3.160 3.162 3.164 3.166
9.2 3.168 3.170 3.172 3.174 3.176 3.178 3.180 3.182 3.184 3.186
9.3 3.188 3.190 3.192 3.194 3.196 3.198 3.200 3.202 3.204 3.206
9.4 3.208 3.210 3.212 3.214 3-215 3.217 3.219 3.221 3.223 3.225
9.5 3227 3.229 3.231 3.233 3-235 3237 3.239 3.241 3-242 3.244
9.6 3.246 3.248 3.250 3.252 3.254 3256 3.258 3.260 3.262 3-264
9.7 - 3.266 3.268 3.269 3-271 3273 3.275 3277 3279 3-281 3.283
9.8 3.285 3.287 3.289 3.291 3293 3.295 3.297 3298 3300 3.302
9.9 3.304 3.305 3.307 3.309 3.311 3.313 3.316 3.318 3320 3321
Me
0.0 0.1 02 03 0.4 0.5 0.6 0.7 0.8 0.9
10 3.32 3.34 3.36 3.37 3.39 3.41 3.43 3.45 3.46 3.48
11 3.50 3.52 3.53 355 3.56 3.58 3.60 3.61 3.63 3.64
12 3.66 3.68 3.69 3.71 3.72 3.74 3.76 3.77 3.79 3.80
13 3.80 3.83 3.85 3.86 3.88 3.89 3.90 3.92 3.93 3.95
14 3.96 3.97 3.99 400 4.02 403 404 406 407 4.09
15 410 4.11 413 414 4.15 417 418 419 4.20 422
16 4.23 4.24 4.25 427 4.28 429 430 431 433 4.34
17 435 436 437 438 439 441 442 443 4.44 445
18 446 447 4.48 4.49 430 452 453 454 4.55 456
19 4.57 458 459 4.60 461 462 4.63 464 4.65 466
O__ COsH
CHARACTERS
Appearance
White or pale yellow, wax-like mass, plates, flakes or
granules.
and enantiomer Solubility
Practically insoluble in water, soluble in ethanol (96 per cent)
C. (RS)-2- [2- [4- [(2-chlorophenyl)phenylmethyl]piperazin-1- and in light petroleum. When mdted, it is misdble with fatty
yl] ethoxy] acetic acid, oils, with liquid paraffin and with mdted wool fat.
IDENTIFICATION
Examine the chromatograms obtained in the assay.
Results The 2 principal peaks in the chromatogram obtained
with the test solution are similar in retention time to the
prindpal peaks in die chromatogram obtained with the
reference solution.
TESTS
Appearance of solution
The solution is clear (2.2.1) and not more intensely coloured
D. 1,4-bis [(4-chlorophenyl)phenylmethyI] piperazine, than reference solution B6 (2.2.2, Method II).
Civ. Dissolve 0.50 g in 20 mL of boiling ethanol (96 per cent) R.
' 0 ' 'COzH Allow to cool.
Melting point (2.2.14)
49 °C to 56 °C.
Acid value (2.5.1)
and enantiomer
Maximum 1.0.
E. (i?S)-2-[2-[2-[4-[(4-chlorophenyI)phenylmethyl]piperazin- Hydroxyl value (2.5.3, Method A)
1-yl] ethoxy] ethoxy]acetic add (ethoxycetirizine), 208 to 228.
ov , co 2h Iodine value (2.5.4, Method A)
Maximum 2.0.
Dissolve 2.00 g in methylene chloride R and dilute to 25 mL
with the same solvent
Saponification value (2.5.6)
Maximum 2.0.
ASSAY
F. 2- [2- [4-(diphenylmethyl)piperazin- 1-yl] ethoxy] acetic add,
Gas chromatography (2.2.28): use the normalisation
CU ^ /-v. /-v .OH procedure.
Test sdudon Dissolve 0.100 g of the substance to be
'N \ / examined in ethanol (96 per cent) R and dilute to 10.0 mL
HW , with the same solvent.
\ — v and enantiomer Reference sdution Dissolve 60 mg of cetyl alcohd CRS and
40 mg of stearyl alcohol CRS in ethanol (96 per cent) R and
G. (RS)-2- [4- [(4-chlorophenyl)phenylmethyl]piperazin-l - dilute to 10 mL with the same solvent. Dilute 1 mL of this
yl]ethan-l-ol. solution to 10 mL with ethanol (96 per cent) R.
PhEur Column:
— sizer. I = 30 m, 0 = 0.32 mm,
— stationary phase, poly(dimethyl)sUoxane R (1 |im).
Carrier gas heliumfor chromatography R.
Content
— stearyl alcohol: m in im u m 40.0 per cent, Detection Flame ionisation.
— sum of the contents of stearyl alcohol and cetyl alcohol:
Irqection 1 jiL.
minimum 90.0 per cent.
2016 Cetostearyl Alcohol 1-491
System suitability: reference solution: to the principal spots in the chromatogram obtained
— resolution: minimum 5.0 between the peaks due to cetyl with reference solution (a);
alcohol and stearyl alcohol. — 2 of the spots in the chromatogram obtained with test
Calculate the percentage contents of C 16H 340 and Ci8H380. solution (b) are similar in position and colour to the
principal spots in the chromatogram obtained with
__________________________________________________________ PhEur
reference solution (b).
6 . Examine the chromatograms obtained in the assay of
cetostearyl alcohol.
Results The 2 principal peaks in the chromatogram obtained
Emulsifying Cetostearyl Alcohol ***** with the test solution are similar in retention time to the
2 principal peaks in the Chromatograms obtained with
(Type A) ***** reference solutions (a) and (b).
(Ph. Ettr. monograph 0801)
C. It gives a yellow colour to a non-luminous flame.
Action and use D. To 0.3 g add 20 mL of anhydrous ethanol R and heat to
Excipient. boiling on a water-bath with shaking. Filter the mixture
immediately, evaporate to dryness and take up the residue in
PhEir__________________________________________________________ 7 mL of water R. To 1 mL of the solution add 0.1 mL of a
DEFINITION 1 g/L solution of methylene blue R, 2 mL of dilute sulfuric
Mixture of cetostearyl alcohol and sodium cetostearyl sulfate. add R and 2 mL of methylene chloride R and shake. A blue
A suitable buffer may be added. colour develops in the lower layer.
Content TESTS
— cetostearyl alcohol: minimum 80.0 per cent (anhydrous Acid value (2.5.1)
substance); Maximum 2.0.
— sodium cetostearyl sulfate: minimum 7.0 per cent Iodine value (2.5.4, Method A)
(anhydrous substance). Maximum 3.0.
CHARACTERS Dissolve 2.00 g in 25 mL of methylene chloride R.
Appearance Saponification value (2.5.6)
White or pale yellow, waxy mass, plates, flakes or granules. Maximum 2.0.
Solubility Water (2.5.12)
Soluble in hot water giving an opalescent solution, practically Maximum 3.0 per cent, determined on 2.50 g.
insoluble in cold water, slightly soluble in ethanol
ASSAY
(96 per cent).
Cetostearyl alcohol
IDENTIFICATION Gas chromatography (2.2.28).
First identification B, C, D \ Internal standard solution Dissolve 0.200 g of
Second identification A, C, D. 1-nonadecanol CRS in anhydrous ethanol R and dilute to
A. Thin-layer chromatography (2.2.27). 100.0 mL with the same solvent.
Test solution (a) Dissolve 0.1 g of the substance to be Test solution Dissolve 0.200 g of the substance to be
examined in 10 mL of trimethylpentane R, heating on a water- examined in 25.0 mL of the internal standard solution.
bath. Shake with 2 mL of ethanol (70 per cent VIV) R and Add 25 mL of water R and shake with 4 quantities, each of
allow to separate. Use the lower layer as test solution (b). 25 mL, of pentane R, adding sodium chloride R , if necessary,
Dilute 1 mL of the upper layer to 8 mL with to facilitate the separation of the layers. Combine the upper
trimethylpentane R. layers, wash with 2 quantities, each of 30 mL, of water R, dry
Test solution (b) Use the lower layer obtained in the over anhydrous sodium sulfate R and filter.
preparation of test solution (a). Reference solution (a) Dissolve 0.100 g of cetyl alcohol CRS in
Reference solution (a) Dissolve 24 mg of cetyl alcohol CRS and 25.0 mL of the internal standard solution. Add 25 mL of
16 mg of stearyl alcohol CRS in 10 mL of trimethylpentane R. water R and shake with 4 quantities, each of 25 m l, of
pentane R, adding sodium chloride R, if necessary, to facilitate
Reference solution (b) Dissolve 20 mg of sodium cetostearyl
the separation of the layers. Combine the upper layers, wash
sulfate R in 10 mL of ethanol (70 per cent VIV) R, heating on
with 2 quantities, each of 30 mL, of water R, dry over
a water-bath.
anhydrous sodium sulfate R and filter.
Plate TLC octadecylsHyl silica gel F 254 plate R.
Reference solution (b) Dissolve 0.100 g of stearyl alcohol CRS
Mobile phase water R, acetone R, methanol R in 25.0 mL of the internal standard solution. Add 25 mL of
(20:40:40 V/VIV). water R and shake with 4 quantities, each of 25 mL, of
Application 10 jxL. pentane R, adding sodium chloride R, if necessary, to facilitate
Development Over 2/3 of the plate. the separation of the layers. Combine the upper layers, wash
Drying In air. with 2 quantities, each of 30 mL, of water R, dry over
anhydrous sodium sulfate R and filter.
Detection Spray with a 50 g/L solution of phosphomolybdic
Column:
acid R in ethanol (96 per cent) R; heat at 120 °C until spots
appear (about 5 min). — material: fused silica;
— size: I = 25 m, 0 = 0.25 mm;
Results:
— stationary phaser. poly(dimethyl)süoxane R (film thickness
— the 2 principal spots in the chromatogram obtained
0.25 pm).
with test solution (a) are similar in position and colour
1-492 Cetostearyl Alcohol 2016
j A3 n ij y 1 Second identification A , C, D.
Ax x —— x x — x 100
A\ A z,y m A. Thin-layer chromatography (2.2.27).
Test solution (a) Dissolve 0.1 g of the substance to be
Az = area of the peak due to stearyl alcohol in the examined in 10 ml. of trimethylpentane R, heating on a water-
chromatogram obtained with the test solution; bath. Shake with 2 mL of ethanol (70 per cent V/V) R and
A Ziy = area of the peak due to stearyl alcohol CRS in the allow to separate. Use the lower layer as test solution (b).
chromatogram obtained with reference solution Dilute 1 mL of the upper layer to 8 mL with
(b); trimethylpentane R.
A\ = area of the peak due to the internal standard in Test solution (b) Use the lower layer obtained in the
the chromatogram obtained with the test preparation of test solution (a).
solution; Reference solution (a) Dissolve 24 mg of cetyl alcohol CRS and
Aj = area of the peak due to the internal standard in 16 mg of stearyl alcohol CRS in 10 mL of trimethylpentane R.
the chromatogram obtained with reference
Reference solution (b) Dissolve 20 mg of sodium
solution (b);
laurilsulfate CRS in 10 mL of ethanol (70 per cent V/V) R,
m = mass of the substance to be examined in the test
solution, in milligrams; heating on a water-bath.
mZjy = mass of stearyl alcohol CRS in reference solution Plate TLC octadecylsUyl silica gel F 254 phue &
(b), in milligrams. Mobile phase water R, acetone R, methanol R
m — mass of the substance to be examined in the test Iodine value (2.5.4, Method A)
solution, in milligrams; Maximum 1.0.
mZiy = mass of stearyl alcohol CRS in reference solution Saponification value (2.5.6)
(b), in milligrams. 135 to 148, determined on 1.0 g.
The percentage content of cetostearyl alcohol corresponds to Heavy metals (2.4.8)
the sum of the percentage contents of cetyl alcohol and Maximum 10 ppm.
stearyl alcohol.
2.0 g complies with test D. Prepare the reference solution
Sodium laurilsulfate using 2 mL of lead standard solution (10 ppm Pb) R
Disperse 0.300 g in 25 mL of methylene chloride R
W ater (2.5.12)
Add 50 mL of water R and 10 mL of dimidium bromide-sulfan
Maximum 0.2 per cent, determined on 10.0 g.
blue mixed solution R. Titrate with 0.004 M benzethonium
chloride, using sonication, heating, and allowing the layers to Total ash (2.4.16)
separate before each addition, until the colour of the lower Maximum 0.2 per cent, determined on 2.0 g.
layer changes from pink to grey. _____________ '____________________________________________ PhEur
1 mL of 0.004 M benzethonium chloride is equivalent to
1.154 mg of sodium laurilsulfate.
LABELLING
The label states, where applicable, the name and Cetrlmide *****
* ★
concentration of any added buffer. (Ph. Eur. monograph 0378) *
__________________________________________________________ PhEur
H3C + CH3
H3C j > N n Br
K , ch3
suitable) into a tared flask and remove the solvent using a ★.**★ ★
rotary evaporator at 40° and then at room temperature at a
Cetyl Alcohol ★ ★
pressure not exceeding 0.7 kPa for 2 hours. The residue (Ph Eur. monograph 0540) *****
weighs not more than 0.4 g.
Non-quatem ised am ines Action and use
To a volume of the solution containing 10 g of cetrimide add Exdpient.
a mixture of 100 mL of propan-2 -ol, 0.1 mL of hydrochloric PhEur.
acid and 20 mL of methanoL Titrate with
0 .1 m tetrabutylammonium hydroxide VS passing a slow current DEFINITION
of nitrogen through the solution and determining the end Mixture of solid alcohols, mainly hexadecan-l-ol (C 16H 34O;
point potentiometrically using a platinum-glass electrode Afr 242.4), of animal or vegetable origin.
system. Inflections in the titration curve indicate (A) Content
neutralisation of excess hydrochloric add and (5) Minimum 95.0 per cent of C 16H 340 .
neutralisation of non-quatemised amine salts. The difference
CHARACTERS
between the volumes corresponding to A and B is not more
than 10 mL (2.4%, calculated as Ci6H 35N). Appearance
White or almost white, unctuous mass, powder, flakes or
Ethanol; Isopropyl alcohol granules.
Carry out one or both of the following methods according to
the dedared alcohol content of die solution being examined. Solubility
Practically insoluble in water, fredy soluble or sparingly
Ethanol Not more than 10.0% v/v, by the method for the
soluble in ethanol (96 per cent). When mdted, it is misdble
determination of ethanol, Appendix VH3 F. Use on-column
with vegetable and animal oils, with liquid paraffin and with
injection and do not heat the injection port. mdted wool fat.
Isopropyl alcohol Not more than 10.0% v/v, by the method
for the determination of ethanol, Appendix VIH F, with the IDENTIFICATION
following modifications. For solution (1) use a solution Examine the chromatograms obtained in the assay.
containing 5.0% v/v of propan-2-ol and 5.0% v/v of propan-1- Results The principal peak in the chromatogram obtained
ol (internal standard). For solution (2 ) vise the solution being with the test solution is similar in retention time to the
examined, diluted with water, if necessary, to contain about prindpal peak in the chromatogram obtained with reference
5.0% v/v of isopropyl alcohol. Maintain the colu m n solution (a).
temperature at 170°, use o n -co lu m n injection and do not TESTS
heat the injection port. Appearance of solution
ASSAY The solution is clear (2.2.1) and not more intensely coloured
Dilute a volume containing 4 g of cetrimide with suffident than reference solution B6 (2.2.2, Method II).
water to produce 100 m L Transfer 25 mL of the solution to Dissolve 0.50 g in 20 mL of boiling ethanol (96 per cent) R.
a separating funnel and add 25 mL of chloroform, 10 mL of Allow to cool.
0.1m sodium hydroxide and 10 mL of a freshly prepared Melting point (2.2.14)
8.0% w/v solution of potassium iodide. Shake well, allow to 46 °C to 52 °C.
separate and discard the chloroform layer. Wash the aqueous
layer with three 10 mL quantities of chloroform and discard Acid value (2.5.1)
the washings. Add 40 mL of hydrochloric acid, cool and titrate Maximum 1.0.
with 0.05m potassium iodate VS until the deep brown colour is Hydroxyl value (2.5.3, Method A)
almost discharged. Add 2 ml, of chloroform and continue the 218 to 238.
titration, with shaking, until the chloroform layer becomes Iodine value (2.5.4, Method A)
colourless. Carry out a blank titration on a mixture of 10 mT. Maximum 2.0.
of the freshly prepared potassium iodide solution, 20 mL of Dissolve 2.00 g in methylene chloride R and dilute to 25 mL
water and 40 mL of hydrochloric acid. The difference between
with the same solvent.
the titrations represents the amount of potassium iodate
required. Each mL of 0.05m potassium iodate FS is equivalent Saponification value (2.5.6)
to 33.64 mg of Ci 7H 38BrN. M axim u m 2.0.
STORAGE ASSAY
Strong Cetrimide Solution should be stored at a temperature Gas chromatography (2.2.28): use the normalisation
above 15°. procedure.
Test solution Dissolve 0.100 g of the substance to be
LABELLING
exam in ed in ethanol (96 per cent) R and dilute to 10.0 mL
The labd states whether Ethanol, Isopropyl Alcohol or both
with the same solvent.
are present and the percentage of cetrimide, wdght in
volume. Reference solution (a) Dissolve 50 mg of cetyl alcohol CRS in
ethanol (96 per cent) R and dilute to 5 mL with the same
solvent.
Reference solution (b) Dissolve 50 mg of stearyl alcohol R in
ethanol (96 per cent) R and dilute to 10 mL with the same
solvent.
Reference solution (c) Mix 1 mL of reference solution (a) and
1 mL of reference solution (b) and dilute to 10 mL with
ethanol (96 per cent) R.
2016 Cetyl Palmitate 1-497
PhEur__________________________________________________________
ASSAY
Dissolve 2.00 g in water R and dilute to 100.0 mL with the
DEFINITION same solvent. Transfer 25.0 mL of the solution to a
Cetylpyridinium chloride contains not less than 96.0 per cent separating funnel, add 25 mL of chloroform R, 10 mL of
and not more than the equivalent of 101.0 per cent of 0.1 M sodium hydroxide and 10.0 mL of a freshly prepared
1-hexadecylpyridinium chloride, calculated with reference to 50 g/L solution of potassium iodide R. Shake well, allow to
the anhydrous substance. separate and discard the chloroform layer. Shake the aqueous
CHARACTERS layer with three quantities, each of 10 mL, of chloroform R
A white or almost white powder, slightly soapy to the touch, and discard the chloroform layers. To the aqueous layer add
soluble in water and in alcohol. An aqueous solution froths 40 mL of hydrochloric add R, allow to cool and titrate with
copiously when shaken. 0.05 M potassium iodaie until the deep-brown colour is almost
discharged. Add 2 mL of chloroform R and continue the
IDENTIFICATION titration, shaking vigorously, until the chloroform layer no
First identification B, D longer changes colour. Carry out a blank titration on a
Second identification A , C, D mixture of 10.0 mL of the freshly prepared 50 g/L solution
A. Dissolve 0.10 g in water R and dilute to 100.0 mL with of potassium iodide R, 20 mL of water R and 40 mL of
the same solvent. Dilute 5.0 mL of this solution to 100.0 mL hydrochloric add R.
with water R. Examined between 240 nm and 300 nm 1 mL of 0.05 M potassium iodate is equivalent to 34.0 mg of
(2.2.25), the solution shows an absorption maxim um at C21H 38C1N.
259 nm and 2 shoulders at about 254 nm and at about __________________________________________________________ PhEur
265 nm. The specific absorbance at the maxim um is 126 to
134, calculated with reference to the anhydrous substance.
B. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with Chalk
cetylpyridinium chloride CRS. Examine the substances in the
Prepared Chalk
solid state.
C. To 5 mL of dilute sodium hydroxide solution R add 0.1 mL CaC03 100.1
of bromophenol blue solution R1 and 5 mL of chloroform R and
Action and use
shake. The chloroform layer is colourless. Add 0.1 mL of
Antacid.
solution S (see Tests) and shake. The chloroform layer
becomes blue. DEFINITION
D. Solution S gives reaction (a) of chlorides (2.3.1). Chalk is a native form of calcium carbonate freed from most
TESTS of its impurities by élutriation and dried. It contains not less
Solution S than 97.0% and not more than 100.5% of CaC03,
Dissolve 1.0 g in carbon dioxide-free water R and dilute to calculated with reference to the dried substance.
100 mL with the same solvent. CHARACTERISTICS
Appearance of solution Chalk absorbs water readily.
Solution S is not more opalescent than reference Practically insoluble in water, slightly soluble in water
suspension 11*(2.2.1) and is colourless (2.2.2, Method II). containing carbon dioxide.
2016 Charcoal 1-499
Arsenic condenser for 1 h, filter and wash the filter with dilute
hydrochloric add R. Evaporate the combined filtrate and
Dissolve 0 .5 g in 5 mL of brominated hydrochloric acid and
dilute to 5 0 mL with water. 25 mL of the resulting solution washings to dryness on a water-bath, dissolve the residue in
0.1 M hydrochloric acid and dilute to 5 0 .0 ml, with the same
complies with the limit testfor arsenic, Appendix VH (4 ppm).
acid.
Heavy metals
Dissolve 1.0 g in 10 mL of 2m hydrochloric acid, add 0.1 mL A ddity or alkalinity
of nitric acid and boil to remove carbon dioxide. Cool, make To 2 .0 g add 4 0 mL of water R and boil for 5 min. Cool,
alkaline with 5m ammonia, filter and wash the precipitate with restore to the original mass with carbon dioxide-free water R
water. Pass 5 mL of hot 2m hydrochloric acid through the
and filter. Reject the first 2 0 mL of the filtrate. To 10 mL of
filter, cool the filtrate, add 0 .5 g of ammonium tkiocyanate and the filtrate add 0 .2 5 ml, of bromothymol blue solution R1 and
0 .2 5 mL of 0.02 M sodium hydroxide. The solution is blue.
extract with two 5 mL quantities of a mixture of equal
volumes of isoamyl alcohol and ether. To the aqueous layer Not more than 0 .7 5 mL of 0.02 M hydrochloric acid is
add 0 .5 g of citric add and dilute to 2 0 mL with water. required to change the colour of the indicator to yellow.
12 mL of the resulting solution complies with limit test A for Add-soluble substances
heavy metals, Appendix VH. Use lead standard solution Maximum 3 per cent.
(2 ppm Pb) to prepare the standard (40 ppm). To 1.0 g add 25 mL of dilute nitric acid R and boil for 5 min.
Chloride Filter whilst hot through a sintered-glass filter (10) (2.1.2)
Dissolve 0.3 g in 2 mL of nitric acid and 10 mL of water, and wash with 10 mL of hot water R. Evaporate the
filter and dilute the filtrate to 30 mL with water. 15 mL of combined filtrate and washings to dryness on a water-bath,
the resulting solution complies with the limit testfor chlorides, add to the residue 1 mL of hydrochloric acid R, evaporate to
Appendix VII (330 ppm). dryness again and dry the residue to constant mass at
1 0 0 -1 0 5 °C. The residue weighs a maximum of 3 0 mg.
Sulfate
Dissolve 0 .2 5 g in 5 .5 mL of 2m hydrochloric add, dilute to Alkali-soluble coloured substances
3 0 mL with water and filter. 15 mL of the resulting solution To 0 .2 5 g add 10 mL of dilute sodium hydroxide solution R
complies with the limit testfor sulfates, Appendix VH (0 .1 2 % ). and boil for 1 min. Cool, filter and dilute the filtrate to
10 mL with water R. The solution is not more intensely
Loss on drying
coloured than reference solution GY4 (2.2.2, Method II).
When dried to constant weight at 105°, loses not more than
1.0% of its weight. Use 1 g. Ethanol (96 per cent) soluble substances
Maximum 0 .5 per cent.
1-500 Chenodeoxycholic Acid 2016
To 2.0 g add 50 mL of ethanol (96 per cent) R and boil under To 10.0 mL of the filtrate add 1.0 g of potassium bromide R
a reflux condenser for 10 min. Filter immediately, cool, and and 20 mL of dilute hydrochloric acid R. Using 0.1 mT. of
dilute to 50 mL with ethanol (96 per cent) R. The filtrate is methyl red solution R as indicator, titrate with 0.0167 M
not more intensely coloured than reference solution Y6 or potassium bromate until the red colour is discharged. Titrate
BY6 (2.2.2, Method II). Evaporate 40 mL of the filtrate to slowly (1 drop every 15 s) towards the end of the titration.
dryness and dry to constant mass at 100-105 °C. The residue Carry out a blank titration using 10.0 mL of the phenazone
weighs a maximum of 8 mg. solution.
Fluorescent substances Calculate the quantity of phenazone adsorbed per 100 g of
In an intermittent-extraction apparatus, treat 10.0 g with activated charcoal from the following expression:
100 mL of cychhexane R1 for 2 h. Collect the liquid and
dilute to 100 mL with cychhexane R l. Examine in ultraviolet 2.353 (a - b)
light at 365 nm. The fluorescence of the solution is not more m
intense than that of a solution of 83 ng of quinine R in
1000 mL of 0.005 M sulfuric acid examined under the same a = number of millilitres of 0.0167 M potassium bromate
conditions. used for the blank;
b = number of millilitres of 0.0167 M potassium bromate
Sulfides
used for the test;
To 1.0 g in a conical flask add 5 mL of hydrochloric acid R l
m = mass in grams of the substance to be examined.
and 20 mL of water R. Heat to boiling. The fumes released
do not turn lead acetate paper R brown. Minimum 40 g of phenazone is adsorbed per 100 g of
activated charcoal, calculated with reference to the dried
Copper
substance.
Maximum 25 ppm.
Atomic absorption spectrometry (2.2.23, Method I). M icrobial contamination
TAMC: acceptance criterion 103 CFU/g (2.6.12).
Test solution Use solution S.
TYMC: acceptance criterion 102 CFU/g (2.6.12).
Reference solutions Prepare the reference solutions using copper
standard solution (0.1 per cent Cu) R and diluting with 0.1 M STORAGE
hydrochloric acid. In an airtight container.
Source Copper hollow-cathode lamp. ___________________________________________________________ PhEur
Wavelength 325.0 nm.
Atomisation device Air-acetylene flame.
Lead
M ax im u m 10 ppm. Chenodeoxycholic Acid /* * %
*. *
Atomic absorption spectrometry (2.2.23, Method I). (Ph. Eur. monograph 1189) *
Test solution Use solution S.
Reference solutions Prepare the reference solutions using lead
standard solution (100 ppm Pb) R and diluting with 0.1 M
hydrochloric acid.
Source Lead hollow-cathode lamp.
Wavelength 283.3 nm; 217.0 nm may be used depending on
the apparatus.
Atomisation device Air-acetylene flame.
Zinc C24H40O4 392.6 474-25-9
Maximum 25 ppm.
Action and use
Atomic absorption spectrometry (2.2.23, Method I). Bile add; treatment of gallstones.
Test solution Use solution S.
PhEur___________________________________________________________-
Reference solutions Prepare the reference solutions using zinc
standard solution (100 ppm Zn) R and diluting with 0.1 M DEFINITION
hydrochloric acid. Chenodeoxycholic add contains not less than 99.0 per cent
Source Zinc hollow-cathode lamp. and not more than the equivalent of 101.0 per cent of 3a,7a-
Wavelength 214.0 nm. dihydroxy-5 ^-cholan-24-oic add, calculated with reference to
the dried substance.
Atomisation device Air-acetylene flame.
Loss on drying (2.2.32) CHARACTERS
Maximum 15 per cent, determined on 1.00 g by drying in an A white or almost white powder, very slightly soluble in
oven at 120 °C for 4 h. water, freely soluble in alcohol, soluble in acetone, slightly
soluble in methylene chloride.
Sulfated ash (2.4.14)
Maximum 5.0 per cent, determined on 1.0 g. IDENTIFICATION
First identification A
Adsorption power
To 0.300 g in a 100 mL ground-glass-stoppered conical flask Second identification B, C
add 25.0 mL of a freshly prepared solution of 0.5 g of A. Examine by infrared absorption spectrophotometry
phenazone R in 50 mL of water R. Shake thoroughly for (2.2.24), comparing with the spectrum obtained with
15 min . Filter and reject the first 5 m l. of filtrate. chenodeoxycholic acid CRS. Examine the substances prepared
as discs using potassium bromide R.
2016 Chitosan Hydrochloride 1-501
B. Examine the chromatograms obtained in the test for (0.5 per cent); any spot apart from the principal spot and any
related substances. The principal spot in the chromatogram spots corresponding to lithocholic aad 5 ursodeoxycholic acid
obtained with test solution (b) is similar in positionj colour and cholic add, is not more intense than the principal spot in
and size to the principal spot in the chromatogram obtained the chromatogram obtained with reference solution (e)
with reference solution (a). (0.25 per cent). The test is not valid unless the
C. Dissolve about 10 mg in 1 mL of sulfuric acid R. chromatogram obtained with reference solution (f) shows two
Add 0.1 mL of formaldehyde solution R and allow to stand for clearly separated prindpal spots.
5 min. Add 5 mL of water R. The suspension obtained is Heavy metals (2.4.8)
greenish-blue. 1.0 g complies with test C for heavy metals (20 ppm).
TESTS Prepare the reference solution using 2 mL of lead standard
solution (10 ppm Pb) R.
Specific optical rotation (2.2.7)
Dissolve 0.500 g in methanol R and dilute to 25.0 mL with Loss on drying (2.2.32)
the same solvent. The specific optical rotation is + 11.0 to Not more than 1.5 per cent, determined on 1.000 g by
+ 13.0j calculated with reference to the dried substance. drying in an oven at 105 °C.
Related substances Sulfated ash (2.4.14)
Examine by thin-layer chromatography (2.2.27), using a Not more than 0.1 per cent, determined on 1.0 g.
suitable silica gel as the coating substance. ASSAY
Test solution (a) Dissolve 0.40 g of the substance to be Dissolve 0.350 g in 50 mL of alcohol R, previously
examined in a mixture of 1 volume of water R and 9 volumes neutralised to 0.2 mL of phenolphthalem solution R.
of acetone R and dilute to 10 mL with the same mixture of Add 50 mL of water R and titrate with 0.1 M sodium
solvents. hydroxide until a pink colour is obtained.
Test solution (b) Dilute 1 mL of test solution (a) to 10 mL 1 mL of 0.1 M sodium hydroxide is equivalent to 39.26 mg of
with a mixture of 1 volume of water R and 9 volumes of C24H 40O 4.
acetone R.
IMPURITIES
Reference solution (a) Dissolve 40 mg of chenodeoxycholic
add CRS in a mixture of 1 volume of water R and 9 volumes
of acetone R and dilute to 10 mL with the same mixture of
solvents.
Reference solution (b) Dissolve 20 mg of lithocholic add CRS in
a mixture of 1 volume of water R and 9 volumes of acetone R
and dilute to 10 mL with the same mixture of solvents.
Dilute 2 mL of the solution to 100 mL with a mixture of
1 volume of water R and 9 volumes of acetone R.
Reference solution (c) Dissolve 20 mg of ursodeoxycholic A. R = H, Rl = OH, R2 = H, R3 = H: 3a-7 ^-dihydroxy-
acid CRS in a mixture of 1 volume of water R and 9 volumes 5P-cholan-24-oic add (ursodeoxycholic add),
of acetone R and dilute to 50 mL with the same mixture of B. R = H, Rl = H, R2 = OH, R3 = OH: 3o,7a,12a-
solvents. trihydroxy-5P-cholan-24-oic add (cholic add),
Reference solution (d) Dissolve 20 mg of cholic add CRS in a
C. R = H, Rl = H, R2 = H, R3 = H: 3a-hydroxy-5p-
mixture of 1 volume of water R and 9 volumes of acetone R cholan-24-oic add (lithocholic add),
and dilute to 100 mL with the same mixture of solvents.
D. R = H, Rl = OH, R2 = H, R3 = OH: 3a,7frl2a-
Reference solution (e) Dilute 0.5 mL of test solution (a) to
trihydroxy-5 (J-cholan-24-oic add (ursocholic add),
20 mL with a mixture of 1 volume of water R and 9 volumes
of acetone R. Dilute 1 mL of the solution to 10 mL with a E. R = H, Rl = H, R2 = H, R3 = OH: 3a,12a-dihydroxy-
mixture of 1 volume of water R and 9 volumes of acetone R. 5P-cholan-24-oic add (deoxycholic add),
Reference solution (f) Dissolve 10 mg of chenodeoxycholic
F. R = H, R1+R2 = = O, R3 = H: 3a-hydroxy-7-oxo-5p-
add CRS in reference solution (c) and dilute to 25 mL with
cholan-24-oic add,
the same solution. G. R = CH3, Rl = OH, R2 = H, R3 = H: methyl 3a,7p-
Apply separately to the plate 5 |iL of each solution. Develop dihydroxy-5 P-cholan-24-oate.
in an unsaturated tank over a path of 15 cm using a mixture __________________________________________________________ PhEur
of 1 volume of glacial acetic add R, 30 volumes of acetone R
and 60 volumes of methylene chloride R. Dry the plate at
120 °C for 10 min. Spray the plate immediately with a
47.6 g/L solution of phosphomofybdic acid R in a mixture of
1 volume of sulfuric add R and 20 volumes of glacial acetic Chitosan Hydrochloride *****
*. *
add R and heat again at 120 °C until blue spots appear on a (Ph. Eur. monograph 1774) *
lighter background. In the chromatogram obtained with test
PhEur__________________________________________________________
solution (a): any spot corresponding to lithocholic acid is not
more intense than the principal spot in the chromatogram DEFINITION
obtained with reference solution (b) (0.1 per cent); any spot Chitosan hydrochloride is the chloride salt of an unbranched
corresponding to ursodeoxycholic acid is not more intense binary heteropolysaccharide consisting of the two units
than the principal spot in the chromatogram obtained with iV-acetyl-D-glucosamine and D-glucosamine, obtained by
reference solution (c) (1 per cent); any spot corresponding to partial deacetylation of chitin normally leading to a degree of
cholic acid is not more intense than the principal spot in the deacetylation of 70.0 per cent to 95.0 per cent Chitin is
chromatogram obtained with reference solution (d) extracted from the shells of shrimp and crab.
1-502 Chitosan Hydrochloride 2016
PhEur___________________________
DEFINITION
2 ,2 ,2 -Trichloroethane-1,1 -diol. ★*★
Chlorambucil ★ ★
Content ★ ★
98.5 per cent to 101.0 per cent. (Ph. Eur. monograph 0137) *****
CHARACTERS
Appearance co 2h
/**★ ★
★
Chloramphenicol ★ ★
(Ph. Eur. monograph 0071) *****
OH
? ' H
D. 2-chloroethyl 4-[4-
[bis(2 -chloroethyl)amino] phenyl] butanoate, H H OH
TESTS ★★*★★
Chloramphenicol Palmitate ★ ★
Acidity or alkalinity
To 0.1 g add 20 mL of carbon dioxide-free water R, shake and (Ph. Eur. monograph 0473) *****
add 0.1 mL of bromothymol blue solution R l. Not more rhan
0.1 mL of 0.02 M hydrochloric add or 0.02 M sodium
hydroxide is required to ch an ge the colour of the indicator.
Specific optical rotation (2.2.7)
Dissolve 1.50 g in ethanol R and dilute to 25.0 mL with the
same solvent. The specific optical rotation is + 18.5 to H OH
+ 20.5.
Related substances C27H 42Q 2N 2O6 561.6 530-43-8
Examine by thin-layer chromatography (2.2.27), using silica
gel G F 254 R as the coating substance. Action and use
Test solution Dissolve 0.10 g of the substance to be examined Antibacterial.
in acetone R and dilute to 10 mL with the same solvent. PhEur.
Reference solution (a) Dissolve 0.10 g of chloramphenicol CRS
in acetone R and dilute to 10 mL with the same solvent. DEFINITION
Chloramphenicol palmitate contains not less than
Reference solution (b) Dilute 0.5 mL of reference solution (a)
98.0 per cent and not more than the equivalent of
to 100 mL with acetone R.
102.0 per cent of (2R,3R)-2-[(dichloroacetyl)amino]-3-
Apply separately to the plate 1 (iL and 20 (iL of the test hydroxy-3-(4-nitrophenyl)propyl hexadecanoate, calculated
solution, 1 (iL of reference solution (a) and 20 fiL of with reference to the dried substance.
reference solution (b). Develop over a path of 15 cm using a
Semi-synthetic product derived from a fermentation product.
mixture of 1 volume of water R, 10 volumes of methanol R
and 90 volumes of chloroform R. Allow the plate to dry in air CHARACTERS
and examine in ultraviolet light at 254 nm. Any spot in the A white or almost white, fine, unctuous powder, practically
chromatogram obtained with 20 pL of the test solution, apart insoluble in water, freely soluble in acetone, sparingly soluble
from the principal spot, is not more intense rhan the spot in in ethanol (96 per cent), very slightly soluble in hexane.
the chromatogram obtained with reference solution (b) It melts at 87 °C to 95 °C.
(0.5 per cent). It shows polymorphism (5.9). The thermodynamically stable
Chlorides (2.4.4) form has low bioavailability following oral administration.
To 1.00 g add 20 m l. of water R and 10 m l. of nitric add R
IDENTIFICATION
and shake for 5 min. Filter through a filter paper previously
A. Examine by thin-layer chromatography (2.2.27), using
washed by filtering 5 mL portions of water R until 5 mL of
TLC sUanised silica gel plate R.
filtrate no longer becomes opalescent on addition of 0.1 mL
of nitric acid R and 0.1 mL of silver nitrate solution R l. 15 m l. Test solution Dissolve 50 mg of the substance to be examined
of the filtrate complies with the limit test for chlorides in a mixture of 1 ml. of 1 M sodium hydroxide and 5 ml. of
(100 ppm). acetone R and allow to stand for 30 min. Add 1.1 mL of 1 M
hydrochloric add and 3 mL of acetone R.
Loss on drying (2.2.32)
Reference solution (a) Dissolve 10 mg of chloramphenicol CRS
Not more than 0.5 per cent, determined on 1.000 g by
drying in an oven at 105 °C. in acetone R and dilute to 5 mL with the same solvent.
Reference solution (b) Dissolve 10 mg of palmitic add R in
Sulfated ash (2.4.14)
acetone R and dilute to 5 mL with the same solvent.
Not more than 0.1 per cent, determined on 2.0 g.
Reference solution (c) Dissolve 10 mg of the substance to be
Pyrogens (2.6.8)
examined in acetone R and dilute to 5 mL with the same
If intended for use in the manufacture of parenteral
solvent.
preparations without a further appropriate procedure for the
removal of pyrogens, it complies with the test for pyrogens. Apply to the plate 4 pL of each solution. Develop over a
Inject per kilogram of the rabbit's mass 2.5 mL of a solution path of 15 cm using a mixture of 30 volumes of a 100 g/L
containing per millilitre 2 mg of the substance to be solution of ammonium acetate R and 70 volumes of ethanol
examined. (96 per cent) R. Allow the plate to dry in air and spray with a
solution containing 0.2 g/L of dichlorcfluorescein R and
ASSAY 0.1 g/L of rhodamine B R in ethanol (96 per cent) R. Allow the
Dissolve 0.100 g in water R and dilute to 500.0 mL with the plate to dry in air and examine in ultraviolet light at 254 nm.
same solvent. Dilute 10.0 mL of this solution to 100.0 mL The chromatogram obtained with the test solution shows
with water R. Measure the absorbance (2.2.25) at the 3 spots corresponding in position to the principal spots in the
maximum at 278 nm. chromatograms obtained with reference solutions (a), (b) and
Calculate the content of C n H ^ C y ^ O s taking the specific (c).
absorbance to be 297. B. Dissolve 0.2 g in 2 mL of pyridine R, add 2 mL of a
STORAGE 100 g/L solution of potassium hydroxide R and heat on a
Store protected from light. If the substance is sterile, store in water-bath. A red colour is produced.
a sterile, airtight, tamper-proof container. C. Dissolve about 10 mg in 5 mL of ethanol (96 per cent) R
_____________________________________ PhEur and add 4.5 mL of dihite sulfuric add R and 50 mg of zinc
powder R. Allow to stand for 10 min and if necessary decant
the supernatant or filter. Cool the solution in iced water and
add 0.5 mL of sodium nitrite solution R. Allow to stand for
2016 Chloramphenicol Sodium Succinate 1-507
2 min and add 1 g of urea R, 2 ml. of strong sodium hydroxide Sulfated ash (2.4.14)
solution Rand 1 ml, of fi-naphthol solution R. A red colour Maximum 0.1 per cent, determined on 1.0 g.
develops. ASSAY
TESTS Dissolve 90.0 mg in ethanol (96 per cent) R and dilute to
Acidity 100.0 mL with the same solvent Dilute 10.0 mL of this
Dissolve 1.0 g in 5 mL of a mixture of equal volumes of solution to 250.0 mL with ethanol (96 per cent) R. Measure
ethanol (96 per cent) R and ether R, warming to 35 °C. the absorbance (2.2.25) of the solution at the m axim u m at
Add 0.2 mL of phertalphthalein solution R. Not more than 271 nm.
0.4 tnl. oi 0.1 M sodmm hydroxide is required to produce a Calculate the content of CjTliæCk^Oôtaking the specific
pink colour persisting for 30 s. absorbance to be 178.
Specific optical rotation (2.2.7) STORAGE
Dissolve 1.25 g in anhydrous ethanol R and dilute to 25.0 mL Protected from light
with the same solvent. The specific optical rotation is
+ 22.5 to + 25.5. IMPURITIES
Free chloramphenicol
Maximum 450 ppm. Dissolve 1.0 g, with gentle heating, in
80 mL of xylene R. Cool and shake with 3 quantities, each of
15 mL, of water R. Dilute the combined aqueous extracts to
50mL with water R and shake with 10 mL of toluene R.
Allow to separate and discard the toluene layer. Centrifuge a
portion of the aqueous layer and measure the absorbance (A)
(.2.2.25) at the maximum at 278 nm using as the A. ( li?,2i?)-2-[(dichloroacetyl)amino]-3-hydroxy-1-
compensation liquid a blank solution having an absorbance (4-nitrophenyl)propyl hexadecanoate (chloramphenicol
not greater than 0.05. palmitate isomer),
Calculate the content of free chloramphenicol in parts per
m illion from the expression:
A x 104
5.96
Related substances
Examine by thin-layer chromatography (2.2.27), using silica
gel G F254 R as the coating substance.
B. ( 1R,2R)-2-[(dichloroacetyl)amino]-1-
Test solution Dissolve 0.1 g of the substance to be examined
(4-nitrophenyl)propane-1,3-diyl bishexadecanoate
in acetone R and dilute to 10 mL with the same solvent. (chloramphenicol dipalmitate).
Reference solution (a) Dissolve 20 mg of chloramphenicol
Ri Eur
palmitate isomer CRS in acetone R and dilute to 10 mL with
the same solvent. Dilute 1 mL of this solution to 10 mL with
acetone R
Reference solution (b) Dissolve 20 mg of chloramphenicol * **
dipalmitate CRS in acetone R and dilute to 10 mL with the Chloramphenicol Sodium ★
★ ★
*
Related substances
Liquid chromatography (2.2.29). Cany out thefollowing
operations protectedfrom bright light and prepare the solutions
immediately before use.
Test solution Dissolve 20.0 mg of the substance to be
examined in the mobile phase and dilute to 100.0 mL with
the mobile phase.
C. (2 -amino-5-chlorophenyl)phenylmethanone
Reference solution (a) Dilute 1.0 mL of the test solution to
(aminochlorobenzophenone).
100.0 mL with the mobile phase. Dilute 2.0 mL of this
_____________________________________________________________ PhEur solution to 10.0 mL with the mobile phase.
Reference solution (b) Dissolve 5 mg of chlordiazepoxide
impurity A CRS in the mobile phase, add 25.0 mL of the test
solution and dilute to 100.0 mL with the mobile phase.
Chlordiazepoxide Hydrochloride ****** Dilute 2.0 mL of this solution to 50.0 mL with the mobile
phase.
*+
(Ph. Eur. monograph 0474) * Reference solution (c) Dissolve 4.0 mg of
aminochlorobenzophenone R in the mobile phase and dilute to
100.0 mL with die mobile phase. Dilute 1.0 mL of this
solution to 100.0 mL with the mobile phase.
Column:
— size. I — 0.15 m, 0 = 4.6 mm,
— stationary phase: octadecylsUyl silica gel for chromatography R
(5 pm).
Mobile phase acetonitrUe R, water R (50:50 V/V).
Flow rate 1.0 mL/min.
C 16H 15C12N30 336.2 438-41-5 Detection Spectrophotometer at 254 nm.
Injection 10 |jL.
Action and use
Run tone 6 times the retention time of chlordiazepoxide.
Benzodiazepine.
Relative retention With reference to chlordiazepoxide
PhEur __ __________________________________________________________ (retention time = about 3.6 min): impurity A = about 0.7;
DEFINITION impurity B = about 2.3; impurity C = about 3.9.
7-Chloro-iV-methyl-5-phenyl-3.fi-1,4-benzodiazepin-2-amine System suitability: reference solution (b):
4-oxide hydrochloride. — resolution: minimum 5.0 between the peaks due to
impurity A and chlordiazepoxide.
Content
99.0 per cent to 101.0 per cent (dried substance). Limits:
— impurities A, B: for each impurity, not more than the area
CHARACTERS of the principal peak in the chromatogram obtained with
Appearance reference solution (a) (0.2 per cent),
White or slightly yellow, crystalline powder. — impurity C: not more than the area of the principal peak
Solubility in the chromatogram obtained with reference solution (c)
Soluble in water, sparingly soluble in ethanol (96 per cent). (0.2 per cent),
It shows polymorphism (5.9). — unspecified impurities: for each impurity, not more than
0.5 times the area of the principal peak in the
IDENTIFICATION chromatogram obtained with reference solution (a)
A. Infrared absorption spectrophotometry (2.2.24). (0.10 per cent),
Comparison chlordiazepoxide hydrochloride CRS. — total: not more than 2.5 times the area of the principal
If the spectra obtained in the solid state show differences, peak in the chromatogram obtained with reference
dissolve 100 mg in 9 mT. of water R and add 1 mL of dilute solution (a) (0.5 per cent),
sodium hydroxide solution R. Extract with 10 mL of methylene — disregard limit: 0.25 times the area of the principal peak in
chloride R in a separating funnel. Evaporate the organic layer the chromatogram obtained with reference solution (a)
and dry the residue obtained at 100-105 °C. Proceed in the (0.05 per cent).
same way with the reference substance. Record new spectra Loss on drying (2.2.32)
using the residues. Maximum 0.5 per cent, determined on 1.000 g by drying
B. Dissolve 50 mg in 5 mL of water R, add 1 mL of dilute in vacuo at 60 °C for 4 h.
ammonia R1, mix, allow to stand for 5 min and filter. Acidify Sulfated ash (2.4.14)
the filtrate with dilute nitric acid R. The solution gives Maximum 0.1 per cent, determined on 1.0 g.
reaction (a) of chlorides (2.3.1).
ASSAY
TESTS Dissolve 0.250 g in 50 mL of water R. Titrate with
Appearance of solution 0.1 M silver nitrate, determining the end-point
The solution is clear (2.2.1) and not more intensely coloured potentiometrically (2.2.20).
than reference solution GY6 (2.2.2, Method IT). 1 mL of 0.1 M silver nitrate is equivalent to 33.62 mg
Dissolve 2.5 g in water R and dilute to 25 mL with the same of C 16H 15C12N 30 .
solvent.
1-512 Chlorhexidine Acetate 2016
STORAGE CHARACTERS
Protected from light. Appearance
IMPURITIES W hite or alm ost white, microcrystalline powder.
Specified impurities: A, B, C. Solubility
Sparingly soluble in water, soluble in ethanol (96 p er cent),
slightly soluble in glycerol and in propylene glycol.
IDENTIFICATION
First identification A.
Second identification B, C, D.
A. Infrared absorption spectrophotom etry (2.2.24).
Comparison chlorhexidine diacetate CRS.
B. Dissolve about 5 mg in 5 m L o f a w arm 10 g/L solution
A. 7-chloro-5-phenyl-1,3-dihydro-2//-1,4-benzodiazepin-2- o f cetrimide R and add 1 m L o f strong sodium hydroxide
one 4-oxide, solution R and 1 m L o f bromine water R. A deep red colour is
produced.
C. Dissolve 0.3 g in 10 m L o f a m ixture o f equal volumes o f
hydrochloric acid R and water R. A dd 40 m L o f water R, filter
if necessary and cool in iced w ater. M ake alkaline to titan
yellow paper R by adding dropwise, and w ith stirring, strong
sodium hydroxide solution R and ad d 1 m l. in excess. Filter,
wash the precipitate with water R until the washings are free
from alkali and recrystallise from ethanol (70 per cent V/V) R.
B. 6-chloro-2-(chloromethyI)-4-phenylquinazoline 3-oxide, D iy at 100-105 °C. T h e residue m elts (2.2.14) a t 132 °C to
136 °C.
D . It gives reaction (a) o f acetates (2.3.1).
TESTS
Impurity P (chloroaniline)
M axim um 500 ppm.
Test solution Dissolve 0.20 g in 25 m L o f water R with
shaking if necessary. Add 1 m L o f hydrochloric add R and
dilute to 30 m L with water R. A dd rapidly and w ith thorough
C. (2-amino-5-chlorophenyl)phenylmethanone mixing after each addition: 5 m L o f a 103 g/L solution o f
(am inochlorobenzophenone). hydrochloric add R, 0.35 m L o f sodium nitrite solution R, 2 m l.
PhEur o f a 50 g/L solution of ammonium sulfamate R, 5 m L o f a
1.0 g/L solution o f naphxhylethylenediamine dihydrochloride R
and 1 m L o f ethanol (96 per cent) R; transfer quantitatively to
a volumetric flask, dilute to 50.0 m L with water R and allow
** + to stand for 30 min.
★ ★
Chlorhexidine Acetate ★ ★ Reference solutions Prepare reference solutions representing
(Chlorhexidine Diacetate, Ph Eicr monograph 0657) ***** respectively 50 ppm , 100 ppm , 200 ppm , 500 ppm and
600 ppm o f chloroaniline R (im purity P) in the test sample as
follows: dilute 1.0 ml-, 2.0 ml-, 4.0 m l-, 10.0 m L and
12.0 mT. o f a solution containing 0.010 g/L o f chloroamline R
(impurity P) in dilute hydrochloric add R to 20 m L with
water R. T h en , add 10 m L o f water R. A dd rapidly and with
- C O jH
thorough mixing after each addition: 5 m L o f a 103 g/L
solution o f hydrochloric add R, 0.35 m L o f sodium nitrite
solution R, 2 m L o f a 50 g/L solution o f ammonium
sulfamate R, 5 m L o f a 1 g/L solution o f
naphxhykthylenediarmne dihydrochloride R and 1 m L o f ethanol
C26H38CI2N10O4 625.6 56-95-1 (96 per cent) R; transfer each solution quantitatively to a
volumetric flask, dilute to 50.0 m L with water R and allow to
Action and use stand for 30 min.
Antiseptic.
M easure th e absorbance (2.2.25) o f each reference solution
Preparation at 556 rnn and plot a calibration curve.
Chlorhexidine Irrigation Solution M easure th e absorbance (2.2.25) o f the test solution at
PhEur.
556 nm . D eterm ine the concentration of chloroaniline from
the calibration curve.
DEFINITION
Related substances
1j 1 '-(H exane-1,6-diyl)bis [5-(4-chlorophenyl)biguanide] Liquid chrom atography (2.2.29). Store the solutions at a
diacetate.
temperature not exceeding 12 °C.
Content
98.0 per cent to 101.0 per cent (dried substance).
2016 Chlorhexidine Acetate 1-513
Test solution Dissolve 0.140 g of the substance to be chromatogram obtained with reference solution (b)
examined in mobile phase A and dilute to 100.0 m L with (0.10 p er cent);
mobile phase A. — total: not more than 0.8 times the area of the principal
Reference solution (a) Dissolve the contents o f a vial of peak in the chromatogram obtained with reference
Chlorhexidine for system suitability CRS (containing solution (b) (0.8 per cent);
impurities A, B, H , I, K, N and O) in 1.0 m L of mobile — disregard limit: 0.05 times the area of the principal peak in
phase A. the chromatogram obtained with reference solution (b)
(0.05 p er cent).
Reference solution (b) Dilute 1.0 m L o f the test solution to
100.0 m L with mobile phase A. L oss on d ry in g (2.2.52)
Column: M aximum 3.5 p er cent, determined on 1.000 g by drying in
— size: I — 0.25 m , 0 = 4.6 mm; an oven at 105 °C.
— stationary phase: end-capped octadecylsüyl silica gel for Sulfated ash (2.4.14)
chromatography R (5 pm); Maximum 0.15 per cent, determined on 1.0 g.
— temperature: 30 °C. ASSAY
Mobile phase: — Dissolve 0.140 g in 100 m L o f anhydrous acetic acid R and
mobile phase A: mix 20 volumes of a 0.1 per cent V/V titrate with 0.1 M perchloric acid. Determine the end-point
solution of trifluoroacetic acid R in acetonitrUe R and potentiometrically (2.2.20).
80 volumes of a 0.1 per cent V/V solution of trifluoroacetic
1 m L of 0.1 M perchloric acid is equivalent to 15.64 mg
add R in water R,
Of C 26H 38CI2N 10O 4.
— mobile phase B: mix 10 volumes o f a 0.1 per cent V/V
solution of trifluoroacetic acid R in water R and 90 volumes IM P U R IT IE S
o f a 0.1 per cent V/V solution of trifluoroacetic acid R in Specified impurities A, H, I, K, N, O, P
acetomtrüe R', Other detectable impurities (the following substances would, if
Time Mobile phase A Mobile phase B present at a sufficient level, be detected by one or other of
(min) (per cent V/V) (per cent V/V) the tests in the monograph. They are limited by the general
0- 2 100 0 acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
2 -3 2 100*80 0 * 20
(2034). It is therefore n o t necessary to identify these
3 2 -3 7 80 20 impurities for demonstration o f compliance. See also 5.10.
37 - 47 80 -»70 2 0 * 30 Control of impurities in substances for pharmaceutical use): B, E,
F, G, M.
4 7 -5 4 70 30
H H
Flow rate 1.0 mL/min.
Detection Spectrophotom eter at 254 nm . TNH
Injection 10 |iL.
Identification of impurities Use the chromatogram supplied Cl
NH NH
with chlorhexidine for system suitability CRS and the
chromatogram obtained with reference solution (a) to
X X
identify the peaks due to impurities A, B, H , I, K, N and O.
Relative retention W ith reference to chlorhexidine (retention
A. l-(4-chlorophenyl)-5-[6-
time = about 35 min): impurity N = about 0.35;
[(cyanocarbamimidoyl) amino]hexyl] biguanide,
impurity B = about 0.36; impurity A = about 0.6;
impurity H = 0.85; impurity O = about 0.90;
impurity I = about 0.91; impurity K = about 1.4. H2N ^ / N N,
System suitability: reference solution (a): TO TNH
— peak-to-valUy ratio: m inim um 2.0, where Hp — height
above the baseline o f the peak due to impurity B and Cl
NH NH
Hv = height above the baseline o f the lowest point o f the
curve separating this peak from the peak due to l X »N x Nx
impurity N . H H
Limits:
— sum cf impurities I and O: not more than 0.4 times the B. 1- [ [6- [ [ [(4-chlorophenyi) carbamimidoyl] carbamimidoyl]
area o f the principal peak in the chromatogram obtained amino]hexyl] carbamimidoyl]urea,
with reference solution (b) (0.4 per cent);
— impurity K: n o t more than 0.3 times the area of the
principal peak in the chromatogram obtained with
reference solution (b) (0.3 per cent);
— impurities A , H, N: for each impurity, n o t more than "X x
H
a
2
0.15 times the area o f the principal peak in the
chrom atogram obtained with reference solution (b)
(0.15 per cent); E. l-( 4-chlorophenyl)guanidine,
— unspecified impurities: for each impurity, not more than
0.1 tim es the area of the principal peak in the
1-514 Chlorhexidine Gluconate Solution 2016
ci H H H
N
H
xo
NH,
*
F. l-(4-chlorophenyi)urea,
H,N.
★
Chlorhexidine Gluconate Solution ★
H H H
H. 1 ,1 '-[iminobis(carbonimidoyliininohexane- 6 , 1-diyl)]bis [5-
(4-chlorophenyl)biguanide], Y y "'
NH NH
I. unknow n structure.
NH NH
H H H
T T H H
O NH
Cl
C 3 4 H 5 4 C I2 N 10 O 14 898 18472-51-0
CI' ' Y ^ S NH NH
Action and use
N N N Antiseptic.
H H H
Preparations
K. 1-(4-chlorophenyl)-3- [[6 - [[ [(4-chlorophenyl) Chlorhexidine G luconate Eye D rops
carbamimidoyl] carbamimidoyi] amino]hexyl] Chlorhexidine Gluconate Gel
carbamimidoyl] urea. Chlorhexidine Irrigation Solution
Chlorhexidine Mouthwash
H H H
Lidocaine and Chlorhexidine Gel
ny ny
NH NH
n'
PhEur.
Cl DEFINITION
NH NH
Aqueous solution o f l,l'-(hexane-l,6-diyl)bis[5-
LX na na n. (4-chlorophenyl)biguanide] di-D-gluconate.
H H H
Content
190 g/L to 210 g/L.
M. 5-(4-chlorophenyl)-5 '-phenyl- 1,1 '-(hexane-1,6-
diyl) dibiguanide, CHARACTERS
Appearance
Almost colourless or pale-yellowish liquid.
Solubility
Miscible with water, with no t m ore than 3 parts o f acetone
and with n o t more than 5 parts o f ethanol (96 per cent).
IDENTIFICATION
First identification A , B.
Second identification B, C, D.
N . l-[6-(carbamimidoylam ino)hexyl]-5- A. Infrared absorption spectrophotom etry (2.2.24).
(4-chlorophenyl)biguanide, Preparation T o 1 m L add 40 m L o f water R, cool in iced
water, make alkaline to titan yellow paper R by adding
dropwise, and with stirring, strong sodium hydroxide solution R
2016 Chlorhexidine Gluconate Solution 1-515
and add 1 m L in excess. Filter, wash the precipitate with naphthylethylenediamine dihydrochloride R and 1 m L of ethanol
water R until the washings are free from alkali and (96 per cent) R] transfer each solution quantitatively to a
recrystallise from ethanol (70 per cent V/V) R. D ry at volumetric flask, dilute to 50.0 m L with water R and allow to
100-105 °C. Examine the residue. stand for 30 min.
Comparison chlorhexidine CRS. M easure the absorbance (2.2.25) o f each reference solution
B. Thin-layer chromatography (2.2.27). at 556 nm and plot a calibration curve.
Test solution Dilute 10.0 m L o f the preparation to be M easure the absorbance (2.2.25) o f the test solution at
examined to 50 m L with water R. 556 nm. D eterm ine the concentration of chloroaniline from
the calibration curve.
Reference solution Dissolve 25 mg of calcium gluconate CRS in
1 m L of water R. R e la te d su b sta n c e s
Plate TLC silica gel plate R. Liquid chromatography (2.2.29). Store the solutions at a
temperature not exceeding 12 °C.
Mobile phase concentrated ammonia R, ethyl acetate R, water R,
ethanol (96 per cent) R (10:10:30:50 VIVIVIV). Test solution Dilute 1.0 m L of the preparation to be examined
to 100.0 m L with mobile phase A.
Application 5 jiL.
Reference solution (a) Dissolve the contents of a vial of
Development Over 1/2 of the plate. chlorhexidine for system suitability CRS (co n taining
Drying At 100 °C for 20 m in and allow to cool. impurities A, B, F, G , H , I, J, K, L, N and O) in 1.0 m L of
Detection Spray with a solution containing 25 g/L of mobile phase A.
ammonium molybdate R and 10 g/L o f cerium sulfate R in dilute Reference solution (b) Dilute 1.0 m L of the test solution to
sulfuric acid R, and heat at 110 °C for about 10 min. 100.0 m L with mobile phase A.
Results T he principal spot in the chromatogram obtained with Column:
the test solution is similar in position, colour and size to the — size. I = 0.25 m , 0 = 4.6 mm;
principal spot in the chromatogram obtained with the — stationary phase: end-capped octadecylsUyl silica gel for
reference solution. chromatography R (5 |im);
C. T o 1 m L add 40 m L o f water R, cool in iced water, make — temperature: 30 °C.
alkaline to titan yellow paper R by adding dropwise, and with Mobile phase:
stirring, strong sodium hydroxide solution R and add 1 m L in — mobile phase A: mix 20 volumes of a 0.1 per cent VIV
excess. Filter, wash the precipitate with water R until the solution of trifluoroacetic acid R in acetomtrUe R and
washings are free from alkali and recrystallise from ethanol 80 volumes of a 0.1 per cent VIV solution of trifluoroacetic
(70 per cent VIV) R. Dry at 100-105 °C. T h e residue melts acid R in water R ;
(2.2.14) at 132 °C to 136 °C. — mobile phase B: mix 10 volumes of a 0.1 per cent VIV
D. T o 0.05 m L add 5 m L of a 10 g/L solution of cetrimide R, solution of trifluoroacetic acid R in water R and 90 volumes
1 m L o f strong sodium hydroxide solution R and 1 m L o f of a 0.1 per cent VIV solution of trifluoroacetic acid R in
bromine water R; a deep red colour is produced. acetomtrUe R-,
TESTS
R elativ e d e n sity (2.2.5) Tune Mobile phase A Mobile phase B
1.06 to 1.07. (min) (per cent V/V) (per cent V/V)
0 -2 100 0
p H (2.2.3)
5.5 to 7.0. 2 - 32 10 0 * 80 0*20
H H
— peak-to-vaRey ratio: minim um 2.0, where Hp = height
above the baseline of the peak due to impurity B and n^ Y '
NH
H v = height above the baseline of the lowest point o f the
curve separating this peak from the peak due to Cl
im purity N . NH NH
Limits: N N N'
— impurity N: not m ore than the area o f the principal peak H H H
in the chrom atogram obtained with reference solution (b)
( 1.0 per cent); A. 1-(4-chlorophenyl)-5- [6-
— impurity H: not more than 0.5 times the area o f the [(cyanocarbamimidoyl)amino]hexyl] biguanide,
principal peak in the chromatogram obtained with
reference solution (b) (0.5 per cent);
h 2n ^ n ^ n ,
— impurities A , J ,K for each impurity, not more than
T T
0.4 times the area of the principal peak in the O NH
chrom atogram obtained with reference solution (b)
(0.4 per cent); Cl
NH NH
— sum of impurities I and O. n o t more than 0.4 times the
area of the principal peak in the chromatogram obtained H H H
w ith reference solution (b) (0.4 per cent);
— impurity G: not m ore than 0.3 times the area of the
B . 1- [ [6- [ [ [(4-chlorophenyl) carbamimidoyi] carbamimidoyl]
principal peak in the chromatogram obtained with
amino]hexyl] carbamimidoyl]urea,
reference solution (b) (0.3 per cent);
— impurities B, F, L, Q. for each impurity, n o t more than
0.2 times the area o f the principal peak in the NH
chrom atogram obtained with reference solution (b)
( 0.2 per cent); N
X NH2
— unspecified impurities: for each impurity, not more than
0.1 times the area of the principal peak in the E. l-(4-chlorophenyi)guanidine,
chrom atogram obtained with reference solution (b)
( 0.10 per cent);
— total: not m ore than 3 times the area o f the principal peak
in the chrom atogram obtained with reference solution (b)
N NH,
(3.0 per cent); H
— disregard limit: 0.05 times the area o f the principal peak in
the chrom atogram obtained with reference solution (b) F . l-(4-chlorophenyi)urea,
(0.05 per cent).
A SSA Y H,N.
H H H
J. l-(4-chlorophenyl)-5-[6-[[4-[(4-chlorophenyl)amino]-6-
nh2
[(lS,2Æ ,3Æ ,4/Q-l,2,3,4,5-pentahydroxypentyl]-l,3,5-triazin-
2-yl] amino] hexyl]biguanide,
P. 4-chloroaniline,
H H H Q. unknown structure.
r r NT NY N' PhEur
o NH
NH NH
Chlorhexidine Hydrochloride ★ ★
U ^H AH -H ★ ★
★ft + ★
(Chlorhexidine Dihydrochloride, Ph. Eur. monograph *
K 1-(4-chlorophenyl)-3- [[6- [[ [(4-chlorophenyl) 0659)
carbamimidoyl] carbamimidoyl] amino] hexyl]
H H
carbamimidoyl] urea,
ny t
NH NH
2HCI
NH NH
I A N
a „
N
a
H H
a V ^ l NH NH
Content
98.0 per cent to 101.0 per cent (dried substance).
U ^ na na n
H H H CHARACTERS
Appearance
M. 5-(4-chlorophenyl)-5 '-phenyl-1 , 1 '-(hexane-1,6- W hite or almost white, crystalline powder.
diyl)dibiguanide, Solubility
very slightly soluble in water, slightly soluble in propylene
H glycol, very slightly soluble in ethanol (96 per cent).
h 2n n ,
YNH IDENTIFICATION
First identification: A, D.
NH NH
Second identification B, C, D
A. Infrared absorption spectrophotometry (2.2.24).
U ^ V l Comparison chlorhexidine dihydrochloride CRS.
B. Dissolve about 5 mg in 5 m L o f a warm 10 g/L solution
N . l-[6-(carbamimidoylamino)hexyl]-5- o f cetrimide R and add 1 m L o f strong sodium hydroxide
(4-chlorophenyl)biguanide, solution R and 1 m L o f bromine water R. A dark red colour is
produced.
C. Dissolve 0.3 g in 10 m L o f a mixture o f equal volumes of
hydrochloric acid R and water R. A dd 40 m L of water R, filter
if necessary and cool in iced water. M ake alkaline to titan
yellow paper R by adding dropwise,* and with stirring, strong
1-518 Chlorhexidine Hydrochloride 2016
sodium hydroxide solution R and add 1 m L in excess. Filter, o f a 0.1 per cent VIV solution o f trifluoroacetic add R in
wash the precipitate with water R until the washings are free acetonitrUe R',
from alkali and recrystallise from ethanol (70 per cent VIV) R.
D ry at 100-105 °C. T h e residue melts (2.2.14) at 132 °C to
Time Mobile phase A Mobile phase B
136 °C.
(min) (per cent V/V) (per cent V/V)
D . It gives reaction (a) of chlorides (2.3.1). 0 -2 100 0
TESTS 2 - 32 100*80 0 *20
Impurity P (chloroanfline)
32 - 37 80 20
M axim um 300 ppm.
Test solution T o 0.20 g add 1 m L of hydrochloric add R, shake 3 7 -4 7 80*70 20*30
for about 30 s, dilute to 30 m L with water R and shake until 47 - 54 70 30
a clear solution is obtained. A dd rapidly and with thorough
mixing after each addition: 5 m L o f a 103 g/L solution o f
hydrochloric acid R, 0.35 m L o f sodium nitrite solution R, 2 m L Flow rate 1.0 mL/min.
of a 50 g/L solution o f ammonium sidfamate R, 5 m l. o f a Detection Spectrophotom eter at 254 nm .
1 g/L solution of naphthykthylenediamine dihydrochloride R and Injection 10 |iL.
1 m L of ethanol (96 per cent) R; transfer quantitatively to a
Identification of impurities Use the chrom atogram supplied
volumetric flask, dilute to 50.0 m L with water R and allow to
w ith chlorhexidine for system suitability CRS and the
stand for 30 min.
chrom atogram obtained with reference solution (a) to
Reference solutions Prepare reference solutions representing identify the peaks due to impurities A, B, H , I, K , N and O.
respectively 50 ppm , 100 ppm , 200 ppm , 300 ppm. and
Relative retention W ith reference to chlorhexidine (retention
500 ppm o f chloroanUme R (impurity P) in the test sample as
tim e = about 35 min): im purity N = about 0.35;
follows: dilute 1.0 m L, 2.0 m L, 4.0 m L, 6.0 m L and
impurity B = about 0.36; impurity A = about 0.6;
10.0 m L of a solution containing 0.010 g/L of chloroani!ine R
impurity H = about 0.85; impurity O = about 0.90;
(impurity P) in dilute hydrochloric acid R to 20 m L with
im purity I = about 0.91; impurity K = about 1.4.
water R. T h en , add 10 m L o f water R. Add rapidly and with
thorough mixing after each addition: 5 m L o f a 103 g/L System suitability: reference solution (a):
solution o f hydrochloric acid i?, 0.35 mT. of sodium nitrite — peak-to-vaUey ratio: minim um 2.0, where Hp — height
solution R, 2 m L o f a 50 g/L solution of ammonium above the baseline o f the peak due to im purity B and
sidfamate R, 5 m L o f a 1 g/L solution of Hv — height above the baseline o f the lowest point o f the
naphthykthylenediamine dihydrochloride R and 1 m L o f ethanol curve separating this peak from the peak due to
(96 per cent) R; transfer each solution quantitatively to a impurity N .
volumetric flask, dilute to 50.0 m L with water R and allow to Limits:
stand for 30 min. — impurity H: no t more than 0.5 times the area of the
principal peak in the chrom atogram obtained with
M easure the absorbance (2.2.25) o f each reference solution
reference solution (b) (0.5 p er cent);
at 556 nm and plot a calibration curve.
— impurity K. not more than 0.4 times the area of the
M easure the absorbance (2.2.25) of the test solution at
principal peak in the chromatogram obtained with
556 nm . D eterm ine the concentration o f chloroaniline from
reference solution (b) (0.4 p er cent);
the calibration curve.
— sum of impurities I and O: not m ore than 0.4 times the
Related substances area o f the principal peak in the chrom atogram obtained
Liquid chrom atography (2.2.29). Store the solutions at a with reference solution (b) (0.4 per cent);
temperature not exceeding 12 °C. — impurities A , N: for each impurity, not m ore than
Test solution Dissolve 0.130 g o f the substance to be 0.15 times the area o f the principal peak in the
examined in mobile phase A and dilute to 100.0 mT. with chrom atogram obtained with reference solution (b)
m obile phase A. (0.15 per cent);
Reference solution (a) Dissolve the contents of a vial o f — unspecified impurities: for each impurity, n o t m ore than
chlorhexidine for system suitability CRS (containing 0.1 times the area o f the principal peak in the
impurities A, B, H , I, K, N and O) in 1.0 m L of mobile chrom atogram obtained with reference solution (b)
phase A. (0.10 per cent);
— total: n o t m ore than the area o f the principal peak in the
Reference solution (b) D ilute 1.0 m L o f the test solution to
chrom atogram obtained with reference solution (b)
100.0 m L w ith mobile phase A.
( 1.0 p er cent);
Column: — disregard limit. 0.05 times the area of the principal peak in
— sizer. I = 0.25 m , 0 = 4.6 mm;
the chrom atogram obtained with reference solution (b)
— stationary phase: end-capped octadecylsUyl silica gel for
(0.05 p er cent).
chromatography R (5 fim);
— temperature: 30 °C. L o ss o n d ry in g (2.2.32)
Maximum 1.0 p er cent, determ ined on 1.000 g by drying in
Mobile phase:
an oven at 105 °C.
— mobile phase A: mix 20 volumes of a 0.1 per cent VIV
solution o f trifluoroacetic acid R in acetonitrUe R and S u lfa te d a s h (2.414)
80 volumes o f a 0.1 per cent VIV solution of trifluoroacetic M aximum 0.1 per cent, determ ined on 1.0 g.
add R in water R; A SSA Y
— mobile phase B: mix 10 volumes o f a 0.1 per cent VIV Dissolve 100.0 m g in 5 m L o f anhydrous formic add R and
solution o f trifluoroacetic add R in water R and 90 volumes add 70 mT. of acetic anhydride R. T itrate with 0.1 Mperchloric
addj determining the end-point potentiometrically (2.2.20).
2016 Chlorhexidine Hydrochloride 1-519
NH
■h « hK Kh
NH
X A „A
N
„A
N N
H H H
H H H
N N N
h 2n ^
TO TNH
n n.
YNH YNH •
Cl
NH NH
NH NH
x A uA 1
N N N
‘N N H H H
H H
^^N^NHz
H 2 CIY '5S NH NH
I A N
a „Na N
E. l-(4-chlorophenyl)guanidine, H H H
N . l-[6-(carbamimidoylamino)hexyI]-5-
0 (4-chlorophenyl)biguanide,
1
N NH2
H 2 H H H
.N ^ ^N,
F. l-(4-chlorophenyl)urea, NH NH
Cl NH NH
I A „NA NA N
H H H
NH NH
N
A..A
N
0.5-(2-chlorophenyl)-5'-(4-chlorophenyl)-l,l '-(hexane-1, 6-
diyl)dibiguanide,
H H
G. 1-(6-aminohexyl)-5-(4-chlorophenyl)biguanide,
^^N H ,
P. 4-chloroaniline.
1-520 Chlorinated Lime 2016
8 - 30 60* 5 40 * 95
3 0 -3 3 5 95
S o lu b ility
System suitability:
— signal-to-noise ratio: minim um 35 for the principal peak in
Practically insoluble in water, soluble in acetonitrile, slightly
the chrom atogram obtained w ith reference solution (b);
soluble in ethanol (96 p er cent).
— peak-to-vaUey ratio: m inim um 1.6, where Hp = height
ID E N T IF IC A T IO N above the baseline of th e peak due to impurity E and
Infrared absorption spectrophotom etry (2.2.24). Hv = height above the baseline o f the lowest point o f the
2016 Chlormadinone Acetate 1-521
ASSAY
Liquid chrom atography (2.2.29) as described in the test for
related substances with the following modifications.
Mobile phase Mobile phase B, mobile phase A (45:55 V/V).
Injection T est solution (b) and reference solution (d).
E. 6-bromo-3,20-dioxopregna-4,6-dien-17-yl acetate,
Run time Twice the retention time of chlormadinone acetate.
Retention time Chlormadinone acetate = about 12 min.
Calculate the percentage content of C 23H 29C 104 taking into
account the assigned content of chlormadinone acetate CRS.
IM P U R IT IE S
Specified impurities A, B, E, G , K
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one o r other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or F. 6^-methyl-3,20-dioxopregn-4-en-17-yl acetate,
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities m substances for pharmaceutical use): C, D,
F ,H ,I ,J ,L
G . 3,20-dioxopregn-4-en-17-yl acetate,
A. 6a-chloro-3,20-dioxopregn-4-en-17-yl acetate,
H . 3-methoxy-20-oxopregna-3,5-dien-17-yl acetate,
B. 2-bromo-6-chloro-3,20-dioxopregna- 1,4,6-trien-17-yl
acetate,
1-522 Chlormethine Hydrochloride 2016
ID E N T IF IC A T IO N
A. Dissolve 50 m g in 5 m L o f water and add 1 m L o f
5m sodium hydroxide. Oily globules are produced which
dissolve on warming.
B. Dissolve 50 m g in 5 m L o f water and add 0.02 m L o f
potassium tetraiodomercurate solution. A cream precipitate is
produced.
C. Melting point, about 108°, A ppendix V A.
I. 6-chloro-3-ethoxy-20-oxopregna-3,5-dien-l 7-yl acetate,
A SSA Y
T o 0.2 g add 15 m L o f I m ethanolic potassium hydroxide and
15 m L o f water and boil under a reflux condenser for
2 hours. Evaporate the solution to h alf its volum e on a w ater
bath, dilute to 150 m L with water, add 3 m l. o f nitric add
and 50 m L o f 0.1m silver nitrate KS, shake vigorously and
filter. W ash the residue with water and titrate the excess of
silver nitrate in the combined filtrate and washings with
0.1m ammonium thiocyanate VS using 1 m L o f ammonium
J. 6-chloro-17-hydroxypregna-4,6-diene-3,20-dione, iron(m) surate solution R2 as indicator. E ach m L o f 0.1m silver
nitrate VS is equivalent to 6.418 m g o f C 5H n C l 2N ,H C l.
o. STO RA G E
Chlorm ethine Hydrochloride should be stored at a
tem perature o f 8 ° to 15°.
L A B E L L IN G
T h e label states th at the contents o f the container are
strongly vesicant.
K. 3>20-dioxopregna-4,6-dien-17-yl acetate,
★*★
★ ★
Chlorobutanol ★ ★
(Chlorobutanol Hemihydrate, Ph. Eux. monograph *****
0383)
h 3c oh
X . ’/.HjO
H3C CCta
L. 6 ß-chloro-3,20-dioxopregn-4-en-17-yl acetate.
Ph Eur GjHyC^OjVi^O 186.5 6001-64-5
A c tio n a n d u se
Disinfectant preservative.
PhEur.
Chlormethine Hydrochloride
D E F IN IT IO N
1, 1,1 -Trichloro- 2-m ethylpropan- 2-ol hem ihydrate.
N' ,HCI
C o n te n t
Me 98.0 per cent to 101.0 per cent (anhydrous substance).
CHARACTERS
C sH nC ljN ^iC l 192.5 55-86-7
A p p e a ra n c e
A c tio n a n d u se W hite or almost white, crystalline pow der or colourless
Cytotoxic alkylating agent. crystals, sublimes readily.
P r e p a r a tio n S o lu b ility
Chlorm ethine Injection Slightly soluble in water, very soluble in ethanol
(96 per cent), soluble in glycerol (85 p er cent).
D E F IN IT IO N mp
Chlorm ethine Hydrochloride is A bout 78 °C (without previous drying).
bis(2-chloroethyl)methylamine hydrochloride. It contains n o t
ID E N T IF IC A T IO N
less than 98.0% and n o t m ore than 101.0% o f
A. Add about 20 m g to a mixture o f 1 m L o f pyridine R and
C s H n C y s m C l.
2 m L o f strong sodium hydroxide solution R. H eat in a water-
C H A R A C T E R IS T IC S bath and shake. Allow to stand. T h e pyridine layer becomes
A white or almost white crystalline powder or mass; red.
hygroscopic; vesicant. B. A dd about 20 m g to 5 m L o f ammoniacal silver nitrate
Very soluble in water. solution R and warm slightly. A black precipitate is formed.
2016 Chlorobutanol 1-523
Column: DEFINITION
— material: glass; Chloroquine phosphate contains n o t less than 98.5 per cent
— size. I = 1.80 m , 0 = 3-4 mm ; and not more than the equivalent o f 101.0 p er cent o f
2016 Chloroquine Sulfate 1-525
iV4-(7-chloroquinolin-4-yl)-N/>N’i-diethylpentane-1,4-diamine obtained w ith the test solution, apart from the principal spot,
bis(dihydrogen phosphate), calculated with reference to the is no t more intense than the spot in the chromatogram
dried substance. obtained with reference solution (a) ( 1.0 p er cent) and not
m ore than one such spot is more intense than the spot in the
CHARACTERS
chromatogram obtained with reference solution (b)
A white or almost white, crystalline powder, hygroscopic,
(0.5 per cent).
freely soluble in water, very slightly soluble in alcohol and in
methanol. H eav y m e ta ls (2.48)
Dissolve 2.0 gin 10 m L o f water R. Add 5 m L of concentrated
I t exists in 2 forms, one o f which melts at about 195 °C and
ammonia R and shake with 40 m L of methylene chloride R.
the other at about 218 °C.
Filter the aqueous layer and neutralise the filtrate with glacial
ID E N T IF IC A T IO N acetic acid R. H eat on a water-bath to elim in ate methylene
First identification B, D. chloride, allow to cool and dilute to 20.0 m L with water R.
Second identification A , C, D. 12 m L o f this solution complies with test A for heavy metals
A. Dissolve 0.100 g in water R and dilute to 100.0 m L with (20 ppm). Prepare the reference solution using lead standard
the same solvent. Dilute 1.0 m l. o f this solution to 100.0 m L solution (2 ppm Pb) R.
with water R. Examined between 210 nm and 370 n m L o ss on d ry in g (2.2.32)
(2.2.25), the solution shows absorption maxima at 220 nm , Maximum 2.0 per cent, determined on 1.000 g by drying in
235 nm , 256 nm , 329 n m and 342 nm . T he specific an oven at 105 °C.
absorbances at the maxima are respectively 600 to 660, ASSAY
350 to 390, 300 to 330, 325 to 355 and 360 to 390.
Dissolve 0.200 g in 50 m L o f anhydrous acetic acid R. T itrate
B. Examine by infrared absorption spectrophotometry with 0.1 M perchloric add determining the end-point
(2.2.24), comparing with the spectrum obtained w ith the potentiometrically (2.2.20).
base isolated from chloroquine sulfate CRS. Record the spectra
1 m L o f 0.1 M perchloric acid is equivalent to 25.79 mg o f
using solutions prepared as follows: dissolve separately 0.1 g
C \ 8H 32CIN 3O 3P 2-
of the substance to be examined and 80 mg of the reference
substance in 10 m L o f water R, add 2 m L o f dilute sodium STORA GE
hydroxide solution R and shake with 2 quantities, each of In an airtight container, protected from light.
20 mT, o f methylene chloride R; combine the organic layers, __________________________________________________________ PhEur
wash with water R, dry over anhydrous sodium sulfate R,
evaporate to dryness and dissolve the residues separately,
each in 2 m L of methylene chloride R.
C. Dissolve 25 m g in 20 m L o f water R and add 8 m L o f ★ ★
picric acid solution R l. T h e precipitate, washed with water R, Chloroquine Sulfate ★ ★
with alcohol R and finally w ith methylene chloride R, melts * * * * *
Chloroquine Sulphate
(2.2.14) at 206-209 °C.
(Ph. Eur. monograph 0545)
D . Dissolve 0.1 g in 10 m L o f water R, add 2 m L o f dilute
sodium hydroxide solution R and shake with 2 quantities, each
of 20 m l , of methylene chloride R. T h e aqueous layer,
acidified by the addition o f nitric acid R, gives reaction (b) of
phosphates (2.3.1). , H2SO4 , h2o
TESTS
S o lu tio n S
Dissolve 2.5 g in carbon dioxide-free water R and dilute to
25 m L with the same solvent.
C w H aO N aO A H aO 436.0 132-73-0
A p p e a ra n c e o f so lu tio n
Solution S is clear (2.2.1) and not more intensely coloured A ctio n a n d u se
than reference solution BY 5 or GY 5 (2.2.2, Method IT). Antiprotozoal (malaria).
p H (2.2.3) P re p a r a tio n s
T he p H o f solution S is 3.8 to 4.3. Chloroquine Sulfate Injection
R elated su b s ta n c e s Chloroquine Sulfate Tablets
Examine by thin-layer chromatography (2.2.27), using silica
gel GF254 R as the coating substance. PhEir.
Test solution Dissolve 0.50 g o f the substance to be examined D E F IN IT IO N
in water R and dilute to 10 m L with the same solvent. Chloroquine sulfate contains n o t less than 98.5 per cent and
Reference solution (a) D ilute 1 m L o f die test solution to n o t more than the equivalent of 101.0 p er cent of
100 m L with water R. iST,-(7-<diloroquinolin-4-yl)-Ari>ZV/-diethylpentane-l,4-diamine
Reference solution (b) D ilute 5 m L o f reference solution (a) to sulfate, calculated with reference to the anhydrous substance.
10 m L with water R. CHARACTERS
Apply to the plate 2 pL o f each solution. Develop over a A white or alm ost white, crystalline powder, freely soluble in
path of 12 cm using a m ixture of 10 volumes of water and in methanol, very slightly soluble in ethanol
diethylamine R, 40 volumes o f cydohexane R and 50 volumes (96 per cent).
of chloroform R. Allow the plate to dry in air. Examine in It melts at about 208 °C (instantaneous method).
ultraviolet light at 254 nm . Any spot in the chromatogram
1-526 Chloroxylenol 2016
ID E N T IF IC A T IO N W a te r (2.5.12)
First identification B, D. 3.0 p er cent to 5.0 per cent, determined on 0.500 g.
Second identification A, C, D. S u lfa te d a s h (2.414)
A. Dissolve 0.100 g in water R and dilute to 100.0 m L with N o t more than 0.1 per cent, determined on 1.0 g.
the same solvent. Dilute 1.0 m L of this solution to 100.0 m L A SSA Y
with water R. Exam ined between 210 nm and 370 nm Dissolve 0.400 g in 50 m L o f anhydrous acetic add R T itrate
(2.2.25), the solution shows absorption maxima at 220 nm , with 0.1 M perchloric acid determining the end-point
235 nm , 256 nm , 329 nm and 342 nm . T he specific potentiometrically (2.2.20).
absorbances a t the maxima are respectively 730 to 810,
1 m L of 0.1 M perchloric add is equivalent to 41.8 mg o f
430 to 470, 370 to 410, 400 to 440 and 430 to 470.
C 18H 28CIN 3O 4S.
B. Examine b y infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with the STO R A G E
base isolated from chloroqume sulfate CRS. Record the spectra Store in an airtight container, protected from light.
using solutions prepared as follows: dissolve separately 0.1 g _________________________________________________________ _ PhEur
of the substance to be examined and of the reference
substance in 10 m L of water R, add 2 m L of dilute sodium
hydroxide solution R and shake with 2 quantities, each of
20 m L, of methylene chloride R", combine the organic layers,
wash with water R, dry over anhydrous sodium sulfate R, Chloroxylenol
evaporate to dryness and dissolve the residues separately each
in 2 m L of methylene chloride R. OH
C. Dissolve 25 m g in 20 m L of water R and add 8 m L of
picric add solution R l. T he precipitate, washed with water R,
with ethanol (96 per cent) R and finally with ether R, melts
(2.2.14) at 206 °C to 209 °C.
D. It gives reaction (a) of sulfates (2.3.1). Cl
TESTS
S o lu tio n S CgHoClO 156.6 88-04-0
Dissolve 2.0 g in carbon dioxide-free water R and dilute to
A ctio n a n d u se
25 m L with the same solvent.
Antiseptic.
A p p e a ra n c e o f so lu tio n
P re p a r a tio n
Solution S is clear (2.2.1) and no t more intensely coloured
Chloroxylenol Solution
than reference solution BY5 or GY 5 (2.2.2, Method II).
p H (2.2.3) D E F IN IT IO N
T he p H of solution S is 4.0 to 5.0. Chloroxylenol is 4-chloro-3,5-xylenoL It contains not less
R e la te d su b s ta n c e s than 98.0% and no t more than 103.0% of CsHgClO.
Examine by thin-layer chromatography (2.2.27), using silica C H A R A C T E R IS T IC S
gel GF254 R as the coating substance. W hite or cream crystals or crystalline powder. It is volatile in
Test solution Dissolve 0.50 g o f the substance to be examined steam.
in water R and dilute to 10 m L with the same solvent. Very slightly soluble in water, freely soluble in ethanol (96%)\
Reference solution (a) Dilute 1 m l. o f the test solution to soluble in ether, in terpenes and in fixed oils. It dissolves in
100 m L with water R. solutions o f the alkali hydroxides.
Reference solution (b) Dilute 5 m L of reference solution (a) to ID E N T IF IC A T IO N
10 m L with water R. T h e infrared absorption spectrum, Appendix II A, is concordant
Apply separately to the plate 2 nL o f each solution. Develop with the reference spectrum of chloroxylenol (RS 055).
over a path of 12 cm using a mixture of 10 volumes of
TESTS
diethylamine R, 40 volumes of cydohexane R and 50 volumes
of methylene chloride R. Allow the plate to dry in air. E xam in e M e ltin g p o in t
114° to 116°, Appendix V A.
in ultraviolet light at 254 nm. Any spot in the chromatogram
obtained with the test solution, apart from the principal spot, T e tra c h lo ro e th y ie n e
is n o t more intense than the spot in the chromatogram Carry out the m ethod for gas chromatography,
obtained with reference solution (a) ( 1.0 per cent) and not Appendix HI B, using the following solutions. Prepare a
more than one such spot is more intense than the spot in the 0 .2 % v/v solution of butanol (internal standard) in methanol
chrom atogram obtained with reference solution (b) (solution A).
(0.5 per cent). ( 1) T o 4 g of the substance being exam ined , add 5 m l . o f
H eav y m e ta ls (2.48) solution A and dilute to 25 m L with methanol.
Dissolve 2.0 g in 10 m L of water R. Add 5 m l, of (2) T o 5 m L of a 0.2% v/v solution of tetrachloroethylene in
concentrated ammonia R and shake with 40 m L of ether R. methanol, add 5 m L o f solution A and dilute to 25 m L with
Filter the aqueous layer and neutralise the filtrate with glacial methanol (equivalent to 0.06488% w/v of tetrachloroethylene in
acetic add R. H eat on a water-bath to elim inate ether, allow methanol).
to cool and dilute to 20.0 m l. with water R. 12 m L o f this
solution complies with test A (20 ppm ). Prepare the
reference solution using lead standard solution (2 ppm Pb) R.
2016 Chlorphenamine Maleate 1-527
CHROMATOGRAPHIC CONDITIONS the flask and allow to stand protected from light for
(a) U se a fused silica capillary column (30 m x 0.53 mm) 15 minutes. A dd 1 g of potassium iodide and 100 m L of water
bonded with a 1 |im film of polyethylene glycol 20,000 (RH- and titrate w ith 0.1m sodium thiosulfate PIS, shaking vigorously
W ax is suitable). and using 1 m L o f starch solution, added towards the end of
(b) U se hydrogen as the carrier gas at 2 m L per m inute. the titration, as indicator. Repeat the operation without the
substance being examined. T he difference between the
(c) Use the gradient conditions described below.
titrations represents the amount o f potassium bromate
(d) Use an inlet temperature o f 240°. required. Each m L o f 0.0167m potassium bromate KS is
(e) Use a flame ionisation detector at a tem perature of 280°. equivalent to 3.915 m g of CgHgClO.
(f) Inject 0.5 |iL o f each solution.
(g) Use a split ratio o f 1:20.
0 -4 isothermal
CO2H
4 -5 70°-»210° linear increase and enantiomer , |
isothermal
n ^CH3 CO2H
5 -1 5 210
i
CH3
15-18 210°->70° linear gradient
oo
re-equilibration
-j
18-20
CaoH^ClNaOi 390.9 113-92-8
A ctio n a n d u se
SYSTEM SUITABILITY Histamine H I receptor antagonist; antihistamine
T he test is n o t valid unless, in the chromatogram obtained
P re p a ra tio n s
with solution ( 2 ), the resolution between the peaks due to
Chlorphenamine Injection
tetrachloroethylene and the internal standard is at least 1.5.
Chlorphenamine Oral Solution
LIMITS
Chlorphenamine Tablets
In the chrom atogram obtained with solution (1), the ratio of
any peak due to tetrachloroethylene to th a t of the internal PhEtr___________________________________________________
standard is n o t greater than the corresponding ratio obtained D E F IN IT IO N
in the chrom atogram obtained with solution (2) (0.4%).
(3/?5)-3-(4-Chlorophenyl)-Ny^-dimethyl-3-(pyridin-2-
R e la te d su b s ta n c e s yI)propan-1-am ine hydrogen (Z)-butenedioate.
Carry out the m ethod for gas chromatography,
C o n te n t
Appendix HI B, using the following solutions in chloroform.
98.0 per cent to 101.0 per cent (dried substance).
( 1) 2 .0 % w/v o f th e substance being examined.
CHARACTERS
(2) 2 .0 % w/v o f the substance being examined and
A p p e a ra n c e
0.040% w/v o f 4-chloro-o-crescl (internal standard).
W hite or alm ost white, crystalline powder.
CHROMATOGRAPHIC CONDITIONS
S olu b ility
(a) Use a glass c o lu m n (1.5 m x 4 mm ) packed with acid- Freely soluble in water, soluble in ethanol (96 per cent).
washed diatomaceous support (80 to 100 mesh) coated with
3% w/w o f polyethylene glycol (Carbowax 20M is suitable). ID E N T IF IC A T IO N
A. Melting point (2.2.14): 130 °C to 135 °C.
(b) Use nitrogen as the carrier gas at 40 m L per minute.
B. Infrared absorption spectrophotometry (2.2.24).
(c) Use isotherm al conditions maintained at 160°.
Comparison chlorphenamine maleate CRS.
(d) Use an inlet tem perature of 200°.
C. Optical rotation (see Tests).
(e) Use a flame ionisation detector at a tem perature of 300°.
(f) Inject 1 |iL o f each solution. TESTS
S o lu tio n S
SYSTEM SUITABILITY
Dissolve 2.0 g in water R and dilute to 20.0 m L with the
T he test is no t valid unless, in the chromatogram obtained same solvent.
w ith solution (2 ), the resolution between th e peaks due to
A p p e a ra n c e o f so lu tio n
chloroxylenol an d 4-chloro-o-cresol is at least 1.5
Solution S is clear (2.2.1) and n o t more intensely coloured
LIMITS than reference solution BY 6 (2.2.2, Method II).
In the chrom atogram obtained with solution (2) the sum o f O p tic a l r o ta tio n (2.2.7)
the areas o f any secondary peaks is not greater than the area of - 0 . 10° to + 0 . 10°, determined on solution S.
the peak due to th e internal standard.
R e la te d su b sta n c e s
A SSAY Liquid chromatography (2.2.29).
Dissolve 70 m g in 30 m L o f glacial acetic acid, add 25 m L of Test solution Dissolve 0.100 g o f the substance to be
0.0167m potassium bromate VS, 20 m L o f a 15% w/v solution examined in the mobile phase and dilute to 100.0 m L with
o f potassium bromide and 10 m L of hydrochloric add, stopper the mobile phase.
1-528 Chlorphenamine Maleate 2016
Reference solution (a) Dilute 0.5 m L o f the test solution to S u lfa te d a s h (2.4.14)
100.0 m L with die mobile phase. M aximum 0.1 per cent, determined on 1.0 g.
Reference solution (b) Dilute 1.0 m L o f reference solution (a) ASSAY
to 10.0 m L with the mobile phase. Dissolve 0.150 g in 25 m L o f anhydrous acetic arid R. T itrate
Reference solution (c) Dissolve 5 mg o f chlorphenamine with 0.1 M perchloric acid, determining the end-point
impurity C CRS in 5 m L of the test solution and dilute to potentiometrically (2.2.20).
50.0 m L with the mobile phase. Dilute 2 m L of this solution 1 m L of 0.1 M perchloric acid is equivalent to 19.54 m g
to 20 m L with the mobile phase. o f C 20H 23C IN 2O 4.
Reference solution (d) Dissolve 5 mg o f 2,2'-dipyridylantine R
STO RA G E
(impurity 8 ) in the mobile phase and dilute to 100 m L with
Protected from light.
the mobile phase.
Reference solution (e) Dissolve the contents of a vial of IM P U R IT IE S
chlorphenamine impurity A CRS in 2 m L of the test solution. Specified impurities: A, B, C, D .
Sonicate for 5 min.
Column:
— size. I = 0.30 m , 0 = 3.9 mm;
— stationary phase. octadecylsUyl silica gel for chromatography R
(10 pm).
^CH3
Mobile phase M ix 20 volumes o f aeetonitrile R and 80 volumes
of a 8.57 g/L solution o f ammonium dihydrogen phosphate R
previously adjusted to p H 3.0 with phosphoric acid R.
Flow rate 1.2 mL/min. A. 2-(4-dilorophenyi)-4-(dimethyiamino)-2-[2-
Detection Spectrophotom eter at 225 nm . (dimediyiamino)ethyl]butanenitrile,
Injection 20 pL.
Run time 3.5 times die retention time o f chlorphenamine.
Relative retention W ith reference to chlorphenamine (retention
time = about 11 min): maleic a d d = about 0 .2 ;
im purity A = about 0.3; impurity B = about 0.4;
(Vo
im purity C = about 0.9; im purity D = about 3.0. B. N-(pyridin- 2-yl)pyridin-2-amine (2,2/-dipyridylamine),
System suitability: reference solution (c):
— resolution: minim nm 1.5 between the peaks due to
impurity C and chlorphenamine.
''= ( H and enantiomer
Limits: „CH3
— correction factors: for the calculation o f contents, multiply
the peak areas o f the following impurities by the
corresponding correction factor impurity A = 1.5;
impurity B = 1.4;
C. (3i?S)-3-(4-chJorophenyi)-AT-methyi-3-(pyridin-2-
— impurity A: not m ore than 0.4 times the area of the
yl)propan - 1-amine,
principal peak in die chromatogram obtained with
reference solution (a) (0.2 per cent);
— impurities B, C, D: for each impurity, not more than
0.2 times the area o f the principal peak in the and enantiomer
chrom atogram obtained with reference solution (a)
(0.1 per cent);
— unspecified impurities: for each impurity, not more than
0.2 times the area o f the principal peak in the
chrom atogram obtained with reference solution (a)
D . (2i?S)-2-(4-chlorophenyl)-4-(dimethylamino)-2-(pyridin-
( 0.10 per cent);
2-yl)butanenitrile.
— total: not m ore than the area of the principal peak in the
chrom atogram obtained with reference solution (a) PhEur
(0.5 per cent);
— disregard limit, the area o f the principal peak in the
chrom atogram obtained with reference solution (b)
(0.05 per cent); disregard the peaks due to the blank and
m aldc ad d .
H eav y m e ta ls (2.4.8)
M aximum 20 ppm .
1.0 g complies with test C. Prepare the reference solution
using 2 m L o f lead standard solution (10 ppm Pb) R.
L oss o n d ry in g (2.2.32)
M áximum 0.5 per cent, determ ined on 1.000 g by drying in
an oven at 105 °C for 4 h.
2016 Chlorpromazine Hydrochloride 1-529
*** ★
★
Chlorpromazine Chlorpromazine Hydrochloride ★ ★
(Fh Eur. monograph 0475) *****
ch3
N
I HCI
CH3
A c tio n a n d u se 69-09-0
D opam ine receptor antagonist; neuroleptic.
A ctio n a n d u se
P re p a r a tio n
D opamine receptor antagonist; neuroleptic.
Chlorpromazine Suppositories
P re p a ra tio n s
D E F IN IT IO N Chlorpromazine Injection
Chlorpromazine is [3-(2-chlorophenothiazin-10-yl)propyl]- Chlorpromazine Oral Solution
dimethylamine. It contains not less than 99.0% and n o t m ore Chlorpromazine Tablets
than 101.0 % o f C 17HX9CIN 2S, calculated with reference to
the dried substance. PhEur_______________________________________
C H A R A C T E R IS T IC S D E F IN IT IO N
A white or creamy white powder or waxy solid. 3-(2-Chloro-l OH-phenothiazin-10-yO -N ^-dim ethylprop an-
Practically insoluble in water, freely soluble in ethanol (96%) 1-amine hydrochloride.
and in ether. C o n te n t
ID E N T IF IC A T IO N 99.0 per cent to 101.0 p er cent (dried substance).
A. T he infrared absorption spectrum, Appendix II A, is CHARACTERS
concordant with the reference spectrum of chlorpromazine A p p e a ra n c e
(RS 056). White or almost white, crystalline powder.
B. Complies with the test for identification ofphenothiazines, S olu b ility
Appendix lH A, using chlorpromazine hydrochloride BPCRS to Very soluble in water, freely soluble in ethanol (96 per cent).
prepare reference solution. It decomposes on exposure to air and light.
TESTS It shows polymorphism (5.9).
M eltin g p o in t
ID E N T IF IC A T IO N
56° to 58°, Appendix V A.
First identification B, D.
R e la te d su b sta n c e s
Second identification A, C, D.
Complies with the test for related substances in phenothiazines,
Appendix IE A, using mobile phase A. A. Ultraviolet and visible absorption spectrophotometry
(2.2.25). Prepare the solutions protected from bright light and
L oss o n d ry in g measure the absorbances immediately.
W hen dried to constant weight over phosphorus pentoxide at a
Test solution Dissolve 50.0 mg in a 10.3 g/L solution of
pressure n o t exceeding 0.7 kPa, loses n o t more than 0.5% o f
hydrochloric add R and dilute to 500.0 m L with the same
its weight. Use 1 g.
solution. Dilute 5.0 m L of the solution to 100.0 m L with a
S u lfa te d a sh 10.3 g/L solution of hydrochloric acid R.
N ot more than 0.1%, Appendix IX A.
Spectral range 230-340 nm.
A SSA Y Absorption maxima At 254 nm and 306 nm.
Dissolve 0.8 g in 300 m L of acetone and carry out M ethod I Specific absorbance at the absorption maximum at 254 nm
for rum-aqueous titration, Appendix VIII A, using 3 m L o f a
890 to 960.
saturated solution of methyl orange in acetone as indicator.
B. Infrared absorption spectrophotometry (2.2.24).
Each m L of 0.1m perchloric acid K5 is equivalent to 31.89 m g
of C 17H 19CIN 2S. Preparation 60 g/L solutions in methylene chloride R using a
0.1 m m cell.
STORAGE
Comparison chlorpromazine hydrochloride CRS.
Chlorpromazine should be protected from light.
C. Identification o f phenothiazines by thin-layer
chromatography (2.3.3): use chlorpromazine hydrochloride CRS
to prepare the reference solution.
D . It gives reaction (b) o f chlorides (2.3.1).
TESTS
p H (2.2.3)
3.5 to 4.5. Carry out the test protected from light and use freshly
prepared solutions.
Dissolve 1.0 g in carbon dioxide-free water R and dilute to
10 m L with the same solvent.
1-530 Chlorpromazine Hydrochloride 2016
* *★
Chlorpropamide ★ ★
★ ★
(Ph. Eur. monograph 1087) *****
° ° ?
i^ V s ' n - ^ n ' ^ - ch’
Cl X J H H
A. 3-(2-chloro-1Oif-phenothiazin-10-yl)-N,N-
dim ethylpropan- 1-amine 5-oxide (chlorpromazine sulfoxide).
A ctio n a n d u se
Inhibition o f A TP-dependent potassium channels
(sulfonylurea); treatm ent of diabetes mellitus.
P re p a r a tio n
Chlorpropamide Tablets
B. N - [3-(2-chloro-lOH-phenothiazin- 10-yl)propyl]-AyV'^N 7 PhEur.
trim ethylpropane-1,3-diamine,
D E F IN IT IO N
Chlorpropamide contains not less than 99.0 per cent and not
m ore than the equivalent of 101.0 per cent of
^ch 3 1 - [(4-chlorophenyl) sulfonyl] -3-propylurea, calculated with
reference to the dried substance.
CHARACTERS
A white o r almost white, crystalline powder, practically
insoluble in water, freely soluble in acetone and in methylene
C . 3-( 1Oif-phenothiazin-10-yl)-N>N'-dimethylpropan-1-amine chloride, soluble in alcohol. It dissolves in dilute solutions of
(promazine), alkali hydroxides.
It shows polymorphism (5.9).
ID E N T IF IC A T IO N
First identification C, D
Second identification A , B, D
A. Melting point (2.2.14): 126 °C to 130 °C.
B. Dissolve 0.10 g in methanolR and dilute to 50.0 m l. with
the same solvent. Dilute 5.0 mL o f the solution to 100.0 m L
D . 3-(2-chloro-1Oif-phenothiazin-10-yl)-N’- m ethylpropan-1- with 0.01 M hydrochloric acid. Dilute 10.0 m l. of the solution
amine (desmethylchlorpromazme). to 100.0 m L w ith 0.01 M hydrochloric acid. Examined
between 220 n m and 350 nm (2.2.25), the solution shows an
absorption maximum at 232 nm. T h e specific absorption at
the maximum is 570 to 630.
C. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
chlorpropamide CRS. Examine the substances prepared as
discs. If the spectra obtained show differences, dissolve the
substance to be examined and the reference substance in
E. 2-chloro - 1OH-phenothiazine,
methylene chloride R, evaporate to dryness and record the new
spectra using the residues.
D . H eat 0.1 g with 2 g of anhydrous sodium carbonate R until
a dull red colour appears for 10 min. Allow to cool, extract
the residue with about 5 m L of water R, dilute to 10 m L
w ith water R and filter. T he solution gives the reaction (a) of
chloride (2.3.1).
TESTS
R e la te d su b sta n c e s
F . 3-(4-chloro - 1OH-phenothiazin-10-yI)-N,iV- Examine by thin-layer chromatography (2.2.27), using a
dimethylpropan - 1-amine. suitable silica gel as the coating substance.
PhEur Test solution Dissolve 0.50 g of the substance to be examined
in acetone R and dilute to 10 m L with the same solvent.
Reference solution (a) Dissolve 15 mg o f
4-chIorobenzenesidfonamide R (chlorpropamide im purity A) in
acetone R and dilute to 100 m L with the same solvent.
1-532 Chlorprothixene Hydrochloride 2016
Reference solution (b) Dissolve 15 m g o f chlorpropamide 1 m L o f 0.1 M sodium hydroxide is equivalent to 27.67 m g of
impurity B CRS in acetone R and dilute to 100 m L with the C 10H 13C lN 2O 3S.
sam e solvent. STO R A G E
Reference solution (c) Dilute 0.3 m l, o f the test solution to Store protected from light.
100 m L with acetone R.
IM P U R IT IE S
Reference solution (d) Dilute 5 m l, o f reference solution (c) to
15 m L with acetone R. o o
* //
Reference solution (e) Dissolve 0.10 g o f the substance to be Ss ,R
N
examined, 5 mg of 4-chlorobenzenesulfonamide R and 5 m g of H
chlorpropamide impurity B CRS in acetone R and dilute to
10 m L with the same solvent.
Apply to the plate 5 jxL of each solution. Develop over a A. R = H : 4-chlorobenzenesulfonamide,
path o f 15 cm using a mixture o f 11.5 volumes of C. R = C O -N H 2: [(4-chlorophenyl)sulfonyl]urea.
concentrated ammonia R, 30 volumes o f cydohexane R,
50 volumes o f methanol R and 100 volumes o f methylene O
chloride R. Allow the plate to dry in a current o f cold air, heat HaC„ A
at 110 °C for 10 min. At the bottom o f a chromatographic
tank, place an evaporating dish containing a mixture of
1 volum e o f hydrochloric add R, 1 volume of water R and B. 1,3-dipropylurea,
2 volumes of a 50 g/L solution of potassium permanganate R, PhEur
close the tank and allow to stand for 15 min. Place the dried
hot plate in the tank and close the tank. Leave the plate in
contact with the chlorine vapour for 2 min. W ithdraw the
plate and place it in a current o f cold air until the excess of J r *★
★ ★
chlorine is removed and an area of coating below the points Chlorprothixene Hydrochloride ★ ★
of application does n o t give a blue colour with a drop of *****
(Ph. Eur. monograph 0815)
potassium iodide and starch sdution R. Spray with potassium
iodide and starch solution R. In the chromatogram obtained
with the test solution: any spot corresponding to im purity A
is n o t more intense than the spot in the chromatogram
obtained with reference solution (a) (0.3 per cent); any spot HCI
corresponding to im purity B is n o t m ore intense than the
spot in the chromatogram obtained with reference
solution (b) (0.3 per cent); any spot, apart from the principal
spot and any spot corresponding to impurity A and B, is not
m ore intense than the spot in the chromatogram obtained C 18H 19C12NS 352.3 6469-93-8
with reference solution (c) (0.3 per cent); not more than two
A c tio n a n d u se
such spots are more intense than the spot in the
Dopam ine receptor antagonist; neuroleptic.
chrom atogram obtained with reference solution (d)
(0.1 per cent). T he test is not valid unless the chromatogram PhEur________________________________________
obtained with reference solution (e) shows three clearly
D E F IN IT IO N
separated spots with approximate Rp values o f 0.4, 0.6 and
(Z)-3-(2-Chloro-9H-thioxanthen-9-ylidene)-NJiV-
0.9 corresponding to chlorpropamide, impurity A and
dim ethylpropan- 1-amine hydrochloride.
im purity B respectively.
H e a v y m e ta ls (2.4.8) C o n te n t
99.0 per cent to 101.0 per cent (dried substance).
Dissolve 2.0 g in a mixture o f 15 volumes of water R and
85 volumes o f acetone R and dilute to 20 m L with the same CHARACTERS
m ixture of solvents. 12 m L o f the solution complies with A p p e a ra n c e
test B for heavy metals (20 ppm ). Prepare the reference W hite or almost white, crystalline powder.
solution using lead standard solution (2 ppm Pb) prepared S o lu b ility
by diluting lead standard solution (100 ppm Pb) R with a Soluble in water and in ethanol (96 per cent), slightly soluble
m ixture of 15 volumes of water R and 85 volumes of
in methylene chloride,
acetone R.
mp: about 220 °C.
L o ss o n d ry in g (2.2.32)
N ot m ore than 0.5 per cent, d eterm ined on 1.000 g by ID E N T IF IC A T IO N
drying in an oven at 100 °C to 105 °C. First identification A , E
S u lfa te d a s h (2.4.14) Second identification B} C, D, E
N o t m ore than 0.1 per cent, determ ined on 1.0 g. A. Infrared absorption spectrophotom etry (2.2.24).
A SSA Y Preparation Dissolve 25 m g in 1 m L of water R. A dd 0.1 m L
Dissolve 0.250 g in 50 m L o f alcohol R previously neutralised o f dilute sodium hydroxide solution R. Shake with 2 m L of
using phenolphthalein solution R1 as indicator and add 25 m L
methylene chloride R. Separate the organic layer and wash with
0.5 m L o f water R. Evaporate the organic layer to dryness
o f water R. T itrate with 0.1 M sodium hydroxide until a pink
colour is obtained. and dry the residue at 40-50 °C. Examine the residues
prepared as discs.
Comparison chlorprothixene hydrochloride CRS.
2016 Chlorprothixene Hydrochloride 1-533
CHARACTERS
.c h 3 Appearance
W hite or yellowish-white powder.
Solubility
Very slightly soluble in water, soluble in acetone and in
methanol, practically insoluble in methylene chloride.
It dissolves in dilute solutions o f alkali hydroxides.
C. (Z)-3-(2-chloro-9i/-thioxanthen-9-ylidene)-N-
methylpropan - 1-amine, It shows polymorphism (5.9).
IDENTIFICATION
Infrared absorption spectrophotom etry (2.2.24).
Comparison chlortalidone CRS.
If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and the reference
substance separately in methanol R, evaporate to dryness and
record new spectra using the residues.
D. (2)-3-(4-chloro-9i/-thioxanthen-9-ylidene)-iy,N- TESTS
dimethylpropan - 1-amine, Acidity
Dissolve 1.0 g with heating in a mixture o f 25 m L of
acetone R and 25 m L of carbon dioxide-free water R. Cool.
Titrate with 0.1 M sodium hydroxide, determ ining the
end-point potentiometrically (2.2.20). N o t more than
0.75 m L o f 0.1 M sodium hydroxide is required.
Related substances
Liquid chromatography (2.2.29).
Solvent mixture Mix 2 volumes o f a 2 g/L solution o f sodium
E. 2-chloro-9i/-thioxanthen-9-one, hydroxide R, 48 volumes of mobile phase B and 50 volumes
o f mobile phase A.
Test solution (a) Dissolve 50.0 m g of the substance to be
examined in the solvent mixture and dilute to 50.0 m L with
the solvent mixture.
Test solution (b) Dilute 10.0 m L of test solution (a) to
100.0 m L with the solvent mixture.
Reference solution (a) Dilute 1.0 m L o f test solution (a) to
F. (£)-3-(2-chloro-9f/-thioxanthen-9-ylidene)-N>iS/’- 100.0 m L with the solvent mixture. D ilute 1.0 m L of this
dim ethylpropan- 1-amine ((£)-isomer). solution to 10.0 m L with the solvent mixture.
PhEur Reference solution (b) Dissolve the contents o f a vial of
chlortalidone for peak identification CRS (containing
impurities B, G and J) in 1 m L of the solvent mixture.
Reference solution (c) Dissolve 50.0 m g o f chlortalidone CRS in
Chlortalidone ★ * the solvent m ixture and dilute to 50.0 m L with the solvent
★ ★ mixture. Dilute 10.0 m L o f this solution to 100.0 m L with
(Ph. Eur. monograph 0546) ***** the solvent mixture.
Column:
— sizer. I = 0.25 m , 0 = 4.6 mm;
— stationary phase: octylsQyl silica gel for chromatography R
and enantiomer
(5 pm);
— temperature: 40 °C.
Mobile phase.
C 14H n a N 204 S — mobile phase A: dissolve 1.32 g of ammonium phosphate R
338.8 77-36-1
in about 900 m L of water R and adjust to p H 5.5 with
Action and use dilute phosphoric add R; dilute to 1000 m L with water R;
Thiazide-like diuretic. — mobile phase B: methanol R2;
Preparations
Chlortalidone Tablets Time Mobile phase A Mobile phase B
Co-tenidone Tablets (min) (per cent V7V) (per cent V/V)
0 - 16 65 35
PhEur.
16-21 65*50 35*50
DEFINITION 2 1 -3 5 50 50
2-Chloro-5- [(1RS)-1-hydroxy-3-oxo-2,3-dihydro-1H-isoindol-
3 5 -4 5 50*65 50*35
1-yl] benzenesulfonamide.
Content
97.0 per cent to 102.0 per cent (dried substance). Flow rate 1.4 mL/min.
2016 Chlortetracycline Hydrochloride 1-535
Relative retention W ith reference to chlortetracycline (retention C 22H 25CIN 2O 8 using the chromatogram obtained with
time = about 26 min): impurity D = about 0.5; reference solution (e) and taking into account the assigned
tetracycline = about 0.6; impurity E = about 0.7; content of tetracycline hydrochloride CRS.
impurity B = about 0.8, impurity A = about 0.86; STORA GE
impurity G = about 0.9; impurity H = about 1.1;
Protected from light. If the substance is sterile, store in a
impurity J = about 1 .4, impurity K = about 1.67;
sterile, airtight, tam per-proof container.
impurity L = about 1.71.
System suitability Reference solution (d): IM P U R IT IE S
— resolution, minimum 1.5 between the peaks due to Specified impurities A, B, D , E, G , H , J, K, L
tetracycline and impurity E; m inim u m 1.5 between the Other detectable impurities (the following substances would, if
peaks due to impurities A and G; m in im u m 1.5 between present at a sufficient level, be detected by one or other of
the peaks due to impurities K and L; if necessary, adjust the tests in the monograph. They are limited by the general
the concentration o f dimethyl sulfoxide in mobile acceptance criterion for other/unspecified impurities and/or
phase A. by die general monograph Substances for pharmaceutical use
Limits. (2034). It is therefore n o t necessary to identify these
— correction factors: for the calculation o f content, multiply impurities for demonstration of compliance. See also 5.10.
the peak areas o f the following impurities by the Control of impurities in substances for pharmaceutical use): C, F,
corresponding correction factor im purity G = 1.4; I.
— impurity J = 0.3; impurity K = 0.4; impurity L = 0.4;
— impurity A: no t m ore than 4 times the area of the OH O OH__ O O
principal peak in the chromatogram obtained with
NHj
reference solution (b) (4.0 per cent);
— impurities B, E: for each impurity, not m ore than the area
of the principal peak in the chromatogram obtained with
reference solution (b) ( 1.0 per cent);
— impurity J: n o t more than 3 times the area o f the principal
peak in the chromatogram obtained with reference A. (4i?,4aS,5aS,6S,12a5)-7-chloro-4-(dimethylamino)-
solution (c) (0.3 per cent); 3,6,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-
— impurities D, G, H, L: for each impurity, n o t m ore than l,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carboxamide
twice the area o f the principal peak in the chromatogram (4-epichlortetracycline),
obtained with reference solution (c) (0.2 per cent);
— impurity K. n o t m ore than 1.5 times the area of the oh o oh o o
principal peak in the chromatogram obtained with
reference solution (c) (0.15 per cent); nh2
— unspecified impurities: for each impurity, no t more than the
1
area o f the principal peak in the chromatogram obtained
a HO H H N—c h 3
with reference solution (c) (0.10 per cent);
h3c
— sum of impurities other than A: n o t more than twice the
area of the principal peak in the chromatogram obtained
with reference solution (b) (2.0 per cent); B. (45,4a5,5a5,65,12a5)-7-chloro-4-(dimethylamino)-
— disregard limit. 0.5 times the area o f the principal peak in 3,6,10,12,12a-pentahydroxy-l, 1 1-dioxo-1,4,4a,5 ,5a, 6,11,12a-
the chromatogram obtained with reference solution (c) octahydrotetracene- 2-carboxamide (demeclocycline),
(0.05 per cent).
oh o oh o o
Heavy metals (2.4.8)
M aximum 50 ppm . nh2
0.5 g complies with test C. Prepare the reference solution
using 2.5 m L o f lead standard solution (10 ppm Pb) R.
Water (2.5.12) h3c
M aximum 2.0 per cent, determined on 0.300 g.
Sulfated ash (2.4.14) C. (4R,4a5,5a5,65,12aS)-4-(dimethylamino)-3,6,10,12,12a-
M axim u m 0.5 p er cent, determined on 1.0 g. pentahydroxy- 1,1 l-dioxo-l,4,4a,5,5a,6,l 1, 12a-
Bacterial endotoxins (2.6.14) octahydrotetracene- 2-carboxamide
Less than 1 IU/m g, if intended for use in the manufacture of (4-epidemethyltetracycline),
parenteral preparations without a further appropriate
procedure for the removal of bacterial endotoxins. OH o 0H...0 o
ASSAY NHj
Liquid chromatography (2.2.29) as described in the test for
1
related substances with the following modification.
HO CH3 H N—CH3
Injection 10 nL o f the test solution and reference solutions (a) h3c
and (e).
Calculate the percentage content of C 22H 24CI2N 2O 8 using D . (4i?,4aS,5aS,6S, 12aS)-4-(dimethylamino)-3,6, 10, 12, 12a-
the chrom atogram obtained with reference solution (a) and pentahydroxy- 6-m eth y l-l,l l-dioxo-l,4,4a,5,5a,6, 11, 12a-
taking into account the assigned content o f chlortetracycline octahydrotetracene-2-carboxamide (4-epitetracycline),
hydrochloride CRS. Calculate the percentage ,content of
1-538 Cholesterol 2016
OH O OH O
OH I
Cl HO H H N-CH 3
h3c
a ch 3 h n - ch 3
h3c
K. (4R,4aS, 12aS)-7-chloro-4-(dimethylamino)-3,10,12,12a-
tetrahydroxy-6-m ethyi-l, 11 -dioxo-1,4,4a,5 ,1 1,12a-
F. (4/?,4aS,6S,8aS)-6-[(lJ?)-7-chloro-4-hydroxy-l-methyl-3- hexahydrotetracene-2-caiboxamide
oxo-1,3-dihydro-2-benzofuran-1-yl]-4-(dimethylamino)-3,8a-
(4-epianhydrochlortetracycline)j
dihydroxy-1, 8-dioxo-1,4,4a,5 ,6,7,8,8a-octahydronaphthalene-
2-carboxamide (4-epiisochlortetracycline)j
Cl CH3 H N-CHs
h, c
ch 3 h n —c h 3
h3c
L. (4-S,4aS, 12aS)-7-chloro-4-(dimethylamino)-3,10,12,1 2 a-
tetrahydroxy-6-methyl- 1,1 l-d io x o -l,4 ,4 a,5 ,l 1, 12a-
G. (45,4a«S36S,8aS)-6-[(lK)-7-chloro-4-hydroxy-1 -methyl-3- hexahydrotetrac ene-2-carboxamide
oxo-1,3-dihydro-2-benzofuran-1-yl] -4-(dimethylamino)-3,8a- (anhydrochlortetracycline).
dihydroxy- 1, 8-dioxo - 1,4,4a,5,6,7,8,8a-octahydronaphthalene- __________________________________________________________ PhEur
2-carboxamide (isochlortetracycline),
OH O OH O
OH I
★*★
★ ★
Cholesterol ★ ★
(Ph. Eur. monograph 0993) *****
d HO CH3 H N—CH3
h3c
H . (45,4a5,5a5,65,12a5)-2-acetyl-7-chloro-4-
(dimethylamino)-3j6,10,12,12a-pentahydroxy-6-methyl-
4a,5a,6,l 2a-tetrahydrotetracene-l, 11 (4//,5/i)-dione (2-acetyl-
2-decarboxamidochlortetracycline),
OH O OH o
OH I
C27H 44O 386.7 57-88-5
I. (4i?,4a£,12aS)-4-(dimethylamino)-3,10,12,12a- DEFINITION
tetrahydroxy- 6 -m ethyl- 1,1 l-dioxo-l,4,4a,5,l 1,12a- Cholest-5-en-3p-ol.
hexahydrotetracene-2-carboxamide Content
(4-epianhydrotetracycline), — cholesterol: m inim um 95.0 per cent (dried substance);
— total sterols: 97.0 per cent to 103.0 per cent (dried
substance).
CHARACTERS
Appearance
White or almost white, crystalline powder.
2016 Cholesterol 1-539
S o lu b ility Column:
Practically insoluble in water, sparingly soluble in acetone — material: fused silica;
and in ethanol (96 per cent). — size. I = 30 m , 0 = 0.25 mm;
It is sensitive to light. — stationary phase, poly (dimethyl)siloxane R (film thickness
0.25 Jim).
ID E N T IF IC A T IO N
Carrier gas helium for chromatography R.
A. M elting point (2.2.14): 147 °C to 150 °C.
Flow rate 2 mL/min.
B. Thin-layer chromatography (2.2.27). Prepare the solutions
immediately before use. Split ratio 1:25.
Test solution Dissolve 10 mg of the substance to be examined Temperature:
in ethylene chloride R and dilute to 5 m L with the same — column: 275 °C;
— injection pore. 285 °C;
solvent.
— detector. 300 °C.
Reference solution Dissolve 10 mg o f cholesterol CRS in ethylene
chloride R and dilute to 5 m L with the same solvent. Detection Flam e ionisation.
Plate TLC silica gel G plate R. Irgecdon 1.0 (iL.
Mobile phase ethyl acetate R, toluene R (33:66 ViV). System suitability: reference solution:
— resolution: minimum 10.0 between the peaks due to
Application 20 jiL. pregnenolone isobutyrate and cholesterol;
Development Immediately, protected from light, over a path of — symmetry factor, m inim u m 0.6 for the peak due to
15 cm. cholesterol.
Drying In air. Calculate the percentage content of cholesterol taking into
Detection Spray 3 times with antimony trichloride solution R; account the assigned content of cholesterol CRS. Calculate the
examine within 3-4 min. percentage content of total sterols by adding together the
Results T h e principal spot in the chromatogram obtained with contents o f cholesterol and other substances with a retention
the test solution is similar in position, colour and si2e to the time less than or equal to 1.5 times the retention time of
principal spot in the chromatogram obtained with the cholesterol. Disregard the peaks due to the internal standard
reference solution. and the solvent.
C . Dissolve about 5 mg in 2 mL of methylene chloride R. STORAGE
A dd 1 m L of acetic anhydride R, 0.01 m L o f sulfuric acid R Protected from light.
and shake. A pink colour is produced which rapidly changes
L A B E L L IN G
to red, then to blue and finally to brilliant green.
T h e label states the source material for the production of
TESTS cholesterol (for example bovine brain and spinal cord, wool
S o lu b ility in e th a n o l (96 p e r c en t) fat or chicken eggs).
In a stoppered flask, dissolve 0.5 g in 50 m L o f ethanol
IM P U R IT IE S
(96 per cent) R at 50 °C. Allow to stand for 2 h. N o deposit
or turbidity is formed.
A cid ity
Dissolve 1.0 g in 10 m L o f ether R, add 10.0 m L o f 0.1 M
sodium hydroxide and shake for about 1 min. H eat gently to
eliminate ether and then boil for 5 min. Cool, add 10 m L of
water R and 0.1 m L o f phenolphthalem solution R as indicator
and titrate with 0.1 M hydrochloric acid until the pink colour
just disappears, stirring the solution vigorously throughout
the titration. Carry out a blank titration. T he difference A. 5a-cholest-7-en-3 (i-ol (lathosterol),
between the volumes of 0.1 M hydrochloric acid required to
change the colour of the indicator in the blank and in the test
is not m ore than 0.3 mL.
L oss o n d ry in g (2.2.32)
Maximum 0.3 per cent, determined on 1.000 g by drying
in vacuo at 60 °C for 4 h.
S u lfa te d a s h (2.4.14)
M axim um 0.1 p er cent, determined on 1.0 g.
A SSA Y B. cholesta-5,24-dien-3|3-ol (desmosterol),
Gas chrom atography (2.2.28).
Internal standard solution Dissolve 0.100 g of pregnenolone
isobutyrate CRS in heptane R and dilute to 100.0 m L with the
same solvent.
Test solution Dissolve 25.0 mg of the substance to be
examined in the internal standard solution and dilute to
25.0 m L w ith the same solution.
Reference solution Dissolve 25.0 m g of cholesterol CRS in the
internal standard solution and dilute to 25.0 m L with the C. 5a-cholesta-7,24-dien-3f}-ol.
same solution.
___________________________________________________ BhEur
1-540 Cholesterol for Parenteral Use 2016
D epending on the animal species of origin, it shows different viscosity range = 1 - 5 m m 2/s, internal diameter of
proportions of 4-sulfate and 6-sulfate groups. tube R = 0.53 m m , volume of bulb C = 5.6 m l , internal
C o n te n t diam eter of tube N — 2.8-3.2 mm) with a funnel-shaped
95 per cent to 105 per cent (dried substance). lower capillary end. Use the same viscometer for all
measurements; measure all outflow times in triplicate.
P R O D U C T IO N T h e test is not valid unless the results do not differ by m ore
T he animals from which chondroitin sulfate sodium is than 0.35 per cent from the mean and if the flow time t\ is
derived m ust fulfil the requirements for the health of animals n o t less than 1.6 x to and not more than 1.8 x to. If this is
suitable for hum an consumption. n o t the case, adjust the concentration of test solution (a) and
CHARACTERS repeat the procedure.
A p p e a ra n c e Calculation of the relative viscosities
W hite or almost white, hygroscopic powder. Since the densities of the chondroitin sulfate solutions and o f
S o lu b ility the solvent are almost equal, the relative viscosities Tin (being
Freely soluble in water, practically insoluble in acetone and T|ri, Tlr2311,3 and 1^ 4) can be calculated from the ratio of the
in ethanol (96 per cent). flow times for the respective solutions (being t1} t^ t3 and
14) to the flow time of the solvent to, but taking into account
ID E N T IF IC A T IO N
the kinetic energy correction factor for the capillary
A. Infrared absorption spectrophotometry (2.2.24). (B = 30 800 s3), as shown below:
Preparation Discs o f potassium bromide R.
Comparison For chondroitin sulfate sodium o f terrestrial
origin use chondroitin sutfate sodium CRS and for chondroitin
sulfate sodium o f marine origine use chondroitin sulfate sodium
id
(marine) CRS. *° .2
co
B. Solution S I (see Tests) gives reaction (b) o f sodium
(2.3.1). Calculation of the concentrations
C. Examine the electropherograms obtained in the test for Calculate the concentration cx (expressed in kg/m3) of
related substances. chondroitin sulfate sodium in test solution (a) using the
Results T h e principal band in the electropherogram obtained following expression:
with the test solution is similar in position to the principal
band in the electropherogram obtained with reference x 100 - h
solution (a). m o p x 10 0 * - ¡o r x 1 0
TESTS
x = percentage content of chondroitin sulfate sodium as
S o lu tio n S I
determined in the assay;
. Dissolve 2.500 g in 50.0 m L of carbon dioxide-free water R.
h = loss on drying as a percentage.
S o lu tio n S2
Calculate the concentration c2 (expressed in kg/m3) of
D ilute 1.0 m L o f solution SI to 10.0 m L with water R.
chondroitin sulfate sodium in test solution (b) using the
p H (2.2.3) following expression:
5.5 to 7.5 for solution S i.
S p ecific o p tic a l ro ta tio n (2.2.7) ci x 0.75
—20 to —30 (terrestrial origin) or - 1 2 to - 1 9 (marine
origin) (dried substance), determined on solution S I. Calculate the concentration c3 (expressed in kg/m3) of
In trin s ic v isco sity chondroitin sulfate sodium in test solution (c) using the
0.01 m 3/kg to 0.15 m 3/kg. following expression:
Test solution (a) Weigh 5.000 g (mop) of the substance to be
examined and add about 80 m l. o f an 11.7 g/L solution of ci x 0.50
sodium chloride R at room temperature. Dissolve by shaking at
room tem perature for 30 min. Dilute to 100.0 m L with an Calculate the concentration c4 (expressed in kg/m3) of
11.7 g/L solution o f sodium chloride R. Filter through a chondroitin'sulfate sodium in test solution (d) using the
m em brane filter (nominal pore size 0.45 pm) and discard the following expression:
first 10 m L. T h e concentration of test solution (a) is only
indicative and m ust be adjusted after an initial measurement ci x 0.25
o f the viscosity o f test solution (a).
Calculation of the intrinsic viscosity
Test solution (b) T o 15.0 m L o f test solution (a) add 5.0 m L
o f an 11.7 g/L solution of sodium chloride R. T h e specific viscosity ti* of the test solution (being r[8l, r\s2i
and t | s4) is calculated from the relative viscosities T|n-
Test solution (c) T o 10.0 m L of test solution (a) add 10.0 m L
(being T|r l , T)r3 and Tir4) according to the following
o f an 1 1.7 g/L solution of sodium chloride R.
expression:
Test solution (d) T o 5.0 m L of test solution (a) add 15.0 m L
o f an 11.7 g/L solution o f sodium chloride R.
Vti - 1
D eterm ine the flow-time (2.2.9) for an 11.7 g/L solution o f
sodium chloride R (to) and the flow times for the 4 test T h e intrinsic viscosity [rj], defined as
solutions (rx, t^ r3 and *4), at 25.00 ± 0.03 °C. Use an
appropriate suspended level viscometer (specifications:
viscometer constant = about 0.005 m m 2/s2, kinematic ( 7)
1-544 Chondroitin Sulfate Sodium 2016
is calculated by linear least-squares regression analysis using Results Any secondary band in the electropherogram obtained
the following equation: with the test solution is no t more intense than the band in
the electropherogram obtained with reference solution (b)
(2 per cent).
— = a x fcn + [17]
C% P ro te in (2.5.33, Method 2)
M aximum 3.0 p er cent (dried substance).
C{ = concentration o f the substance to be examined Test solution Dilute 1.0 m L o f solution S I to 50.0 m L w ith
expressed in kg/m3; 0.1 M sodium hydroxide.
&h = H uggins' constant. Reference sdudons Dissolve about 0.100 g o f bovine albumin R,
R e la te d s u b s ta n c e s accurately weighed, m 0.1 M sodium hydroxide and dilute to
Electrophoresis (2.2.31). 50.0 m L with the same solvent. C a n y out all additional
Buffer solution A (0.1 M barium acetate p H 5.0). Dissolve dilutions using 0.1 M sodium hydroxide.
25.54 g o f barium acetate R in 900 m L o f water R. Adjust to C h lo rid e s (2.4.4)
p H 5.0 with glacial acetic add R and dilute to 1000.0 m L M aximum 0.5 per c e n t
w ith water R. D ilute 1 m L o f solution S2 to 15 m L with water R. D o n o t
Buffer solution B (1 M barium acetate p H 5.0). Dissolve add diluted nitric ad d . Prepare the standard using 5 m L of
255.43 g o f barium acetate R in 900 m L o f water R. Adjust to chloride standard solution (5 ppm CO R and 10 m L of water R.
p H 5.0 with glacial acetic acid R and dilute to 1000.0 m L H eav y m e ta ls (2.4.8)
w ith water R. M aximum 20 ppm .
Staining solution Dissolve 1.0 g of tohddine blue R and 2.0 g o f 1.0 g complies w ith test C . Prepare th e reference solution
sodium chloride R in 1000 m L of 0.01 M hydrochloric add using 2 m L o f lead standard solution (10 ppm Pb) R.
Filter.
L oss o n d ry in g (2.2.32)
Test solution Prepare a 30 mg/mL solution of the substance to M aximum 12.0 p er cent, determ ined on 1.000 g by drying in
be examined in water R.
an oven at 105 °C for 4 h.
Reference solution (a) Prepare a 30 m g/m L solution o f
M ic ro b ia l c o n ta m in a tio n
chondroitin sulfate sodium CRS in water R.
TAM C: acceptance criterion 103 C FU /g (2.6.12).
Reference solution (b) D ilute 2.0 m L o f reference solution (a)
TY M C: acceptance criterion 102 C FU /g (2.6.12).
to 100.0 m L with water R.
Absence o f Staphylococcus aureus (2.6.13).
Reference solution (c) M ix equal volumes o f reference
solution (b) and water R. Absence o f Pseudomonas aeruginosa (2.6.13).
Procedure Allow the electrophoresis support to cool the plate Absence o f Escherichia cdi (2.6.13).
to 10 °C. Pre-equilibrate the agarose gel for 1 min in buffer Absence o f Salmonella (2.6.13).
solution A. Remove excess liquid by careful decanting. Absence o f bile-tolerant gram-negative bacteria (2.6.13).
D ry the gel for approximately 5 min. Place 400 m L o f buffer
A SSAY
solution B into each o f the containers o f the electrophoresis
equipm ent. Transfer 1 pL o f each solution to the slots of the Test solution (a) Weigh 0.100 g (m{) o f the substance to be
agarose gel. Pipette a few millilitres o f a 50 per cent V/V examined, dissolve in water R and dilute to 100.0 m L with
solution o f glycerol R onto the cooled plate of the the same solvent.
electrophoresis equipm ent and place the gel in the middle of Test solution (b) Dilute 5.0 m L o f test solution (a) to
the ceramic plate. Place a wick, saturated with buffer 50.0 m L with water R
solution B, at the positive and negative sides of the agarose Reference solution (a) Weigh 0.100 g (mo) o f chondroitin sulfate
gel. Ensure that there is good contact between the sodium CRS, previously dried as described in the test for loss
electrophoresis buffer and th e agarose gel. Perform the on drying, dissolve in water R and dilute to 100.0 m L with
electrophoresis under the following conditions: 75 mA/gel, the same solvent
resulting in a voltage o f 100-150 V (maximum 300-400 V) Reference solution (b) D ilute 5.0 m L o f reference solution (a)
for a gel o f about 12 cm x 10 cm. Carry out the to 50.0 m L with water R.
electrophoresis for 12 min. Place the gel in a mixture
Titrcmt solution (a) W dgh 4.000 g o f cetylpyridimurn chloride
consisting o f 10 volumes o f anhydrous ethand R and
monohydrate R and dilute to 1000 m L with water R.
90 volumes o f buffer solution A for 2 min. C any o u t the
electrophoresis for 20 min. Place the gel in a mixture Titrcmt sdution (b) Weigh 1.000 g o f cetylpyridxrdian chloride
consisting o f 30 volumes o f anhydrous ethand R and monohydrate R and dilute to 1000 m L with water R.
70 volumes o f buffer solution A for 2 min. Carry o u t the Perform either visual o r photom etric titration as follows:
electrophoresis for 20 m in. Stain the gel in the staining Visual titration Titrate 40.0 m L o f reference solution (a) and
solution for 10 min. D estain the gel for 15 min under 40.0 m L o f test solution (a) with titrant solution (a).
running tap water followed by 10-15 m in with wetter R until T h e solution becomes turbid. At the end point, the liquid
the band in the electropherogram obtained with reference appears d e ar, with an ahnost-white pred p itate in suspension.
solution (c) is visible. Allow the gel to dry. T h e predpitate is more apparent if 0.1 m L o f a 1 p er cent
System suitability: solution o f methylene blue R is added before starting the
— the electropherogram obtained w ith reference solution (c) titration. T h e predpitated partides are more apparent against
shows a visible band; the blue background.
— the band in the electropherogram obtained w ith reference Photometric titration T itrate 50.0 m L o f reference solution (b)
solution (b) is clearly visible and similar in position to the and 50.0 m L o f test solution (b) with titrant solution (b).
band in the electropherogram obtained with reference T o determine the end point, use a suitable autotitrator
solution (a).
2016 Chorionic Gonadotrophin 1-545
S p ecific a b so rb a n c e (2.2.25)
Chymotrypsin ?***
18.5 to 22.5, determined at the absorption maximum at
(Ph. Eitr. monograph 0476) * 281 nm ; maximum 8, determ ined at the absorption
minim um at 250 nm.
9004-07-3
Dissolve 30.0 m g in 0.001 M hydrochloric add and dilute to
A c tio n a n d u se 100.0 m L with the same acid.
Proteolytic enzyme. T ry p sin
Substrate solution T o 98.5 m g o f tosylargmine methyl ester
PhEur—_________________________________________
hydrochloride R, suitable for assaying trypsin, add 5 m L o f
D E F IN IT IO N tris(hydroxymethyl) aminomethane buffer solution pH 8.1 R and
Chymotrypsin is a proteolytic enzyme obtained by the swirl to dissolve. Add 2.5 m L of methyl red mixed solution R
activation o f chrymotrypsinogen extracted from the pancreas and dilute to 25.0 m L w ith water R.
of b eef (Bos taurus L .). It has an activity o f not less than Test solution T ransfer to a depression in a white spot-plate
5.0 microkatals per milligram. In solution it has maximal 0.01 m l, o f tris(kydroxymethyl)aminomethane buffer solution
enzymic activity at about p H 8; the activity is reversibly pH 8.1 R and 0.1 m L o f solution S. Add 0.2 m L o f the
inhibited at p H 3, the p H at which it is most stable. substrate solution.
P R O D U C T IO N Reference solution A t the sam e time and in the same m anner .
T he animals from which chymotrypsin is derived m ust fulfil as for the test solution, prepare a solution using the
the requirem ents for the health o f animals suitable for hum an substance to be examined to which n o t more than
consum ption. Furtherm ore, the tissues used shall n o t include 1 per cent m/m o f trypsin BRP has been added.
any specified risk material as defined by any relevant Start a timer. N o colour appears in the test solution within
international or, where appropriate, national legislation. 3-5 min after the addition o f the substrate solution. A purple
T he m ethod o f m anufacture is validated to demonstrate that colour is produced in the control solution.
the product, if tested, would comply with the following test. L oss o n d ry in g (2.2.32)
H is ta m in e C2.6.10) N o t m ore than 5.0 per cent, determ ined on 0.100 g by
N ot m ore than 1 |ig (calculated as histamine base) per drying at 60 °C at a pressure no t exceeding 0.7 kPa for 2 h.
5 microkatals o f chymotrypsin activity. Before carrying out
A SSAY
the test, heat the solution o f the substance to be examined on
T he activity o f chymotrypsin is determ ined by comparing the
a w ater-bath for 30 min.
rate at which it hydrolyses acetykyrosine ethyl ester R with the
CHARACTERS rate at which chymotrypsin BRP hydrolyses the same substrate
A p p e a ra n c e under the same conditions.
White or alm ost white, crystalline or amorphous powder, Apparatus U se a reaction vessel o f about 30 m L capacity
hygroscopic if amorphous. provided with:
S o lu b ility — a device that will maintain a tem perature o f
Sparingly soluble in water. 25.0 ± 0.1 °C;
— a stirring device, for example a magnetic stirrer;
ID E N T IF IC A T IO N
— a lid with holes for the insertion o f electrodes, the tip o f a
A. D ilute 1 m L of solution S (see Tests) to 10 m L with
burette, a tube for the admission o f nitrogen and the
water R .J n a depression in a white spot-plate, mix 0.05 m L introduction o f reagents.
of this solution with 0.2 m L of the substrate solution.
An autom atic or m anual titration apparatus may be used.
A purple colour develops.
F o r the latter, the burette is graduated in 0.005 m L and the
Substrate solution T o 24.0 m g of acetykyrosine ethyl ester R add p H m eter is provided with a wide scale and glass-calomel or
0.2 m L o f ethanol (96 per cent) R and swirl to dissolve.
glass-silver-silver chloride electrodes.
Add 2.0 m L o f 0.067 Mphosphate buffer solution pH 7.0 R
and 1 m L o f methyl red mixed solution R and dilute to Test solution Dissolve 25.0 m g o f the substance to be
10.0 m L w ith water R. examined in 0.001 M hydrochloric add and dilute to
250.0 m L with the same acid.
B. D ilute 0.5 m L of solution S to 5 m L with water R.
Add 0.10 m L o f a 20 g/L solution of Reference solution Dissolve 25.0 m g o f chymotrypsin BRP in
tosylphenylaUtnylchloromethane R in ethanol (96 per cent) R. 0.001 M hydrochloric acid and dilute to 250.0 m L with the
same ad d .
Adjust to p H 7.0 and shake for 2 h. In a depression in a
white spot-plate, mix 0.05 m L o f this solution with 0.2 m l. Store the solutions at 0-5 °C. W arm 1 m L o f each solution
o f the substrate solution (see Identification test A). N o colour to about 25 °C over 15 min and use 50 (iL of each solution
develops within 3 min o f mhring, (corresponding to about 25 nanokatals) for each titration.
Carry out the titration in an atmosphere o f nitrogen. Transfer
TESTS
10.0 m L o f 0.01 M caldum chloride solution R to the reaction
S o lu tio n S
vessel and, while stirring, add 0.35 m L o f 0.2 M acetykyrosine
Dissolve 0.10 g in carbon dioxide-free water R and dilute to ethyl ester R. W hen the tem perature is steady at
10.0 m L with the same solvent.
25.0 ± 0.1 °C (after about 5 m in), adjust to p H 8.0 exactly
A p p e a ra n c e o f so lu tio n with 0.02 M sodium hydroxide. A dd 50 (iL o f the test solution
Solution S is n o t m ore opalescent than reference (equivalent to about 5 pg o f the substance to be examined)
suspension II (2.2.1). and start a timer. M aintain at p H 8.0 by the addition o f
p H (2.2.3) 0.02 M sodium hydroxide, noting the volume added every
3.0 to 5.0 for solution S. 30 s. Calculate the volume o f 0.02 M sodium hydroxide used
per second between 30 s and 210 s. Carry out a titration in
2016 Ciclesonide 1-547
the sam e m anner u sin g the reference solution and calculate B. Examine the chromatograms obtained in the assay.
the volume of 0.02 M sodium hydroxide used per second. Results T h e principal peak in the chromatogram obtained
Calculate the activity in microkatals per milligram using the with the test solution is similar in retention time and size to
following expression: the principal peak in the chromatogram obtained with
reference solution (a).
m' x V TESTS
x A
m xV ’ Related substances
Liquid chromatography (2.2.29).
mass o f the substance to be examined, in
Test solution Dissolve 50.0 mg of die substance to be
m illigram s;
examined in anhydrous ethanol R and dilute to 50.0 m L with
m' mass o f chymotrypsin BRP, in milligrams;
the same solvent.
V volume of 0.02 M sodium hydroxide used per
second by the test solution; Reference solution (a) Dissolve 50.0 mg o f ciclesonide CRS in
V volume of 0.02 M sodium hydroxide used per anhydrous ethanol R and dilute to 50.0 m L with the same
second by the reference solution; solvent.
A activity o f chymotrypsin BRP, in microkatals per Reference solution (b) Dissolve 3 mg o f ciclesonide
milligram. impurity B CRS, 3 mg o f ciclesonide impurity C CRS and 5 mg
o f ciclesonide containing impurity A CRS in anhydrous ethanol R
STORAGE and dilute to 10.0 m L with the same solvent.
In an airtight container at 2 °C to 8 °C, protected from light.
Reference solution (c) Dissolve 50 m g o f the substance to be
L A B E L L IN G examined in anhydrous ethanol R, add 1.0 m L of reference
The label states: solution (b) and dilute to 50.0 m L with anhydrous ethanol R.
— the quantity o f chymotrypsin and the total activity in Reference solution (d) Dilute 1.0 m L of the test solution to
microkatals per container; 100.0 m L with anhydrous ethanol R. Dilute 1.0 m L of this
— for the amorphous substance, that it is hygroscopic. solution to 10.0 m L with anhydrous ethanol R.
PhEur Column:
— size. I = 0.25 m, 0 = 4.6 mm;
— stationary phase: phenylsQyl silica gel for chromatography R
(5 nm);
— temperature. 60 °C.
*****
Ciclesonide ★ ★ Mobile phase water R, anhydrous ethanol R (38:62 VIV).
(Ph. Eur. monograph 2703) **★** Flow rate 1.0 mL/min.
Detection Spectrophotom eter at 243 nm .
Injection 20 (xL of the test solution and reference solutions (c)
and (d).
Run time 2.2 times the retention time of ciclesonide.
Identification of impurities Use the chromatogram obtained
with reference solution (c) to identify the peaks due to
impurities A, B and C.
Relative retention W ith reference to ciclesonide (retention
time = about 16 min): impurity B = about 0.4;
impurity C = about 0.9; impurity A = about 1.4.
C 32H 44O7 540.7 12654447-6 System suitability, reference solution (c):
— resolution: m inim u m 1.5 between the peaks due to
A ctio n a n d u se impurity C and ciclesonide.
Glucocorticoid.
Calculation of percentage contents:
PhEur____________ — for each impurity, use the concentration of ciclesonide in
reference solution (d).
D E F IN IT IO N
Limits:
(2'R)-2 '-Cyclohexyl- 11 J}-hydroxy-3,20-dioxo-l 6$H- — impurity A: maximum 1.0 per cent;
[l,3]dioxolo[4',5/:16,17]pregna-l,4-dien-21-yl
— impurities B, C: for each impurity, maximum
2-methylpropanoate.
0.15 per cent;
C o n te n t — unspecified impurities:, for each impurity, maximum
98.0 per cent to 102.0 per cent (anhydrous substance). 0.10 per cent;
CHARACTERS — total of unspecified impurities:, maximum 0.2 per cent;
A p p e a ra n c e — total: maximum 1.2 per cent;
W hite or yellowish-white, crystalline powder. — reporting threshold: 0.05 per cent.
W a te r (2.5.12)
Ciclopirox ★**★ ★
M axim um 0.5 per cent, determined on 0.500 g. ★ ★
(Ph. Em. monograph 1407) *****
S u lfa te d a s h (2.4.14)
M axim um 0.1 per cent, determined on 1.0 g.
A SSA Y
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modifications.
Injection T est solution and reference solution (a).
Run time 1.6 times the retention time o f dclesonide.
System suitability: reference solution (a):
— symmetry factor, maximum 2.2 for the peak due to C 12H 17N O 2 207.3 29342-05-0
dclesonide.
A c tio n a n d u se
Calculate the percentage content o f C 32H 44O 7 taking into
Antifungal.
account the assigned content of dclesonide CRS.
IM P U R IT IE S PhEtr.
Specified impurities A, B, C. D E F IN IT IO N
6-Cyclohexyl-1-hydroxy-4-methylpyridin-2 (l/f)-o n e .
C o n te n t
98.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
A p p e a ra n c e
W hite o r yellowish-white, crystalline powder.
S o lu b ility
Slightly soluble in water, fredy soluble in anhydrous ethanol
and in methylene chloride.
ID E N T IF IC A T IO N
A. (2 'S)-2 '-cydohexyl -1 1^-hydroxy-3,20-dioxo-16 $H-
[1,3] dioxolo[4 ',5 ': 16,17]pregna-l,4-dien-21-yl First identification B.
2-m ethylpropanoate (S-epimer of dclesonide), Second identification A , C.
A. M d tin g point (2.2.14): 140 °C to 145 °C.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison ciclopirox CRS.
C. Thin-layer chromatography (2.2.27).
Test solution Dissolve 20 mg of the substance to be examined
in methanol R and dilute to 10 m L with the same solvent.
Reference solution Dissolve 20 m g o f ciclopirox CRS in
methanol R and dilute to 10 m L with the same solvent.
B. (2 'R)-2 '-cyclohexyl-1 113,21-dihydroxy- 16$H- Plate TLC silica gel F254 plate R.
[ 1,3] dioxolo [4 ',5 ' : 16 ,17] pregna-1,4-diene-3,20-dione,
Pretreatment Before use, predevelop with the mobile phase
until the solvent front has migrated to the top o f the plate.
Allow to dry in air for 5 min.
Mobile phase concentrated ammonia R, water R, ethanol
(96 per cent) R (10:15:75 V/V/V).
Application 10 (iL.
Development Over 2/3 o f the plate.
Drying In air for 10 mini
Detection A Examine in ultraviolet light at 254 nm .
Results A T h e prindpal spot in the chrom atogram obtained
C. (2'i?)-2'-[(l.RS)-cyclohex-3-enyi]-l 1 fi-hydroxy-3,20- w ith the test solution is sim ilar in position and size to the
dioxo-16 $H- [1,3] dioxolo [4 ',5 ' : 16,17] pregna-1,4-dien-21-yl principal spot in the chromatogram obtained with the
2-methylpropanoate. reference solution.
PhEtr
Detection B Spray with a 20 g/L solution of ferric chloride R in
anhydrous ethanol R.
Results B T h e prindpal spot in the chrom atogram obtained
with the test solution is similar in position, colour and size to
the principal spot in the chromatogram obtained with the
reference solution.
2016 Ciclopirox 1-549
ID E N T IF IC A T IO N — total: not m ore than 1.5 times the area o f the principal
A Infrared absorption spectrophotom etry (2.2.24). peak in the chrom atogram obtained with reference
Comparison ciclosporin CRS. solution (b) (1.5 p er cent);
— disregard limit. 0.05 times the area o f the principal peak in
8 . Examine the chromatograms obtained in the assay.
the chromatogram obtained with reference solution (b)
Results The principal peak in the chromatogram obtained (0.05 per cent).
with the test solution is similar in retention time to the
principal peak in the chrom atogram obtained with reference H e av y m e ta ls (2.48)
solution (a). M axim um 20 ppm .
T h e residue obtained in the test for loss on drying complies
TESTS
with test C. Prepare the reference solution using 2 m L of
A p p e a ra n c e o f so lu tio n lead standard solution (10 ppm Pb) R.
T he solution is clear (2.2.1) and n o t m ore intensely coloured
than reference solution Y5, BY 5 or R 7 (2.2.2, Method II). L oss o n d ry in g (2.2.32)
M axim um 2.0 per cent, determ ined on 1.000 g at 60 °C at a
Dissolve 1.5 g in anhydrous ethanol R and dilute to 15 m L
pressure not exceeding 15 P a for 3 h.
with the same solvent.
B a c te ria l e n d o to x in s (2.6.14)
S pecific o p tic a l ro ta tio n (2.2.7)
Less than 0.84 IU/m g, if intended for use in the m anufacture
- 1 9 3 to - 1 8 5 (dried substance).
of parenteral preparations w ithout a further appropriate
Dissolve 0.125 g in methanol R and dilute to 25.0 m L with procedure for the removal o f bacterial endotoxins. Dissolve
the same solvent. 50 m g of the substance to be examined in a mixture of
R e la te d su b s ta n c e s 280 m g of ethanol (96 per cent) R and 650 m g o f
Liquid chrom atography (2.2.29). polyoxyethylated castor où R and dilute to the required
Solvent mixture acetonitrUe R, water R (50:50 VIV). concentration using water for BET.
Test solution Dissolve 30.0 m g of the substance to be A SSA Y
examined in the solvent mixture and dilute to 25.0 m L with Liquid chromatography (2.2.29) as described in the test for
the solvent mixture. rd a te d substances with the following modifications.
Reference solution (a) Dissolve 30.0 m g of ciclosporin CRS in Injection T est solution and reference solution (a).
the solvent m ixture and dilute to 25.0 m L with the solvent System suitability: reference solution (a):
mixture. — repeatability, m axim u m rdative standard deviation o f
Reference solution (b) D ilute 2.0 m L o f reference solution (a) 1.0 p er cent after 6 injections.
to 200.0 m L with the solvent mixture. Calculate the percentage content o f C ^ H m N n O ^ taking
Reference solution (c) Dissolve the contents o f a vial of into account the assigned content o f ddosporin CRS.
ciclosporin for system suitability CRS in 5.0 m l. of the mobile STORAGE
phase.
In an airtight container, protected from light. If the substance
Column: is sterile, store in a sterile, airtight, tam per-proof container.
— size: I = 0.25 m , 0 = 4 mm ;
— stationary phase: octadecylsilyl silica gel for chromatography R IM P U R IT IE S
(3-5 pm);
R
— temperature: 80 °C.
r*- Ala—D-Ala—MeLeu—MeLeu—MeVal - N H CH3
T he column is connected to the injection port by a steel l 4 5 y *1 V .
capillary tube about 1 m long, having an internal diam eter o f *- MeLeu * Val-«- MeLeu * M e G l y * Abu—/ /> |
0.25 m m and m aintained at 80 °C. n 7 OH k,
Mobile phase phosphoric acid R, 1,1-dimethyleihyl methyl ether R,
acetonitrile R, water R (0.1:5:43:52 VIVIV/V).
Flow rate 1.5 mlVmin. A. different d dosporins [difference from d d o sp o rin
Detection Spectrophotom eter a t 210 nm . (R = C H 3: d d o sp o rin A)]: d d o sp o rin B [7-L-Ala];
Injection 20 p L o f the test solution and reference solutions (b) d d o sp o rin C [7-L-Thr]; d d o sp o rin D [7-L-VaI];
and (c). d d o sp o rin E [5-L-VaIJ; d d o sp o rin G [7-(l-2-
Run time 1.7 times the retention time o f ddosporin. aminopentanoyl)]; d d o sp o rin H [5-o-MeVal]; d d o sp o rin L
System suitability: reference solution (c): [R = H ]; d d o sp o rin T [4-L-Leu]; d d o sp o rin U [11-L-Leu];
— retention time, d d o sp o rin = 25 min to 30 min; d d o sp o rin V [1-L-Abu],
if necessary, adjust the ratio o f acetonitrile to water in the
ch3
mobile phase;
(-"-Ala—o-AJa— MeLeu— MeLeu— MeVal- N H CH3
— peak-to-vaUey ratio: minim um 1.4, where Hp = h d g h t
above the baseline o f the peak due to d d o sp o rin U and L
*- MeLeu * Val MeLeu * MeGly Abu -
Hv = hdgjht above the baseline of the lowest point o f the
curve separating this peak from the peak due to
ddosporin; if necessary, adjust the ratio of
1, 1-dimethylethyl methyl ether to acetonitrile in the
mobile phase. B. [6-[(2S,3£,4£)-3-hydroxy-4-m ethyl-2-
Limits: (methylamino)octanoic ad d ]]d d o sp o rin A,
any impurity: for each impurity, n o t m ore than 0.7 times C . isoddosporin A.
the area o f the principal peak in the chromatogram
PhEur
obtained with reference solution (b) (0.7 per cent);
2016 Cilastatin Sodium 1-553
S o lu b ility 78-88 30 70
Very soluble in w ater and in methanol, slightly soluble in
anhydrous ethanol, very slightly soluble in dimethyl sulfoxide,
practically insoluble in acetone and in methylene chloride. Flow rate 2.0 mL/min.
ID E N T IF IC A T IO N Detection Spectrophotometer at 210 nm.
A. Specific optical rotation (see Tests). Injection 20 jiL.
B. Infrared absorption spectrophotometry (2.2.24). Identification of impurities Use the chromatogram supplied
Comparison cilastatin sodium CRS. with cilastatin for system suitability 1 CRS and the
chromatogram obtained with reference solution (b) to
C. It gives reaction (a) of sodium (2.3.1).
identify the peaks due to impurities A, B, E, F, G (epimer 2)
TESTS and H ; use the chromatogram supplied with cilastatin for
S o lu tio n S system suitability 2 CRS and the chromatogram obtained with
Dissolve 1.0 g in carbon dioxide-free water R and dilute to reference solution (c) to identify the peaks due to impurities
100 m L with the same solvent. C and G (epimer 1); use the chromatogram obtained with
A p p e a ra n c e o f so lu tio n reference solution (d) to identify the peak due to impurity D .
Solution S is clear (2.2.1) and not more intensely coloured Relative retention W ith reference to cilastatin (retention
than reference solution Y 6 (2.2.2, Method II). time = about 50 min): impurity E = about 0 .2; impurity A
(epimer 1) = about 0.60; impurity A (epimer
p H (2.2.3)
2) = about 0.62; impurity D = about 0.9;
6.5 to 7.5 for solution S.
impurity F = about 0.98; impurity G (epimer 1) = about
S pecific o p tical ro ta tio n (2.2.7) 1.02; impurity G (epimer 2) = about 1.05;
+ 41.5 to + 44.5 (anhydrous substance). impurity H = about 1.06; impurity B = about 1.17;
Dissolve 0.250 g in a mixture of 1 volume o f hydrochloric impurity C = about 1.23.
acid R and 120 volumes of methanol R, then dilute to System suitability:
25.0 m L with the same mixture of solvents. — peak-to-vaUey ratio: minimum 10, where Hp = height
R e la te d su b sta n c e s above the baseline o f the peak due to impurity F and
Liquid chromatography (2.2.29). Prepare the solutions Hv = height above the baseline of the lowest point of the
immediately before use. curve separating this peak from the peak due to cilastatin
Test solution Dissolve 32 mg of the substance to be examined in the chromatogram obtained w ith reference solution (b);
in water R and dilute to 20.0 m L with the same solvent — peak-to-vaUey ratio: minimum 2.5, where Hp = height
above the baseline of the peak due to impurity G
Reference solution (a) Dilute 1.0 m L of the test solution to
(epimer 1) and Hv = height above the baseline of the
100.0 m L with water R. Dilute 1.0 m L of this solution to
lowest point of the curve separating this peak from the
10.0 m L with water R.
peak due to cilastatin in the chromatogram obtained with
Reference solution (b) Dissolve 3 mg of cilastatin for system reference solution (c).
suitability 1 CRS (co n taining impurities A, B, E, F , G
Calculation of percentage contents:
(epimer 2) and H ) in water R and dilute to 2.0 m L with the
— for all impurities, use the concentration of cilastatin in
same solvent.
reference solution (a);
1-554 Cilastatin Sodium 2016
— correction factors: multiply the peak areas o f the following Ru = ratio of the area o f the solvent peak to the area of
impurities by the corresponding correction facto r the propanol peak in the chrom atogram obtained
im purity C = 1.3; im purity E = 3.3; impurity G with the test solution;
(epimer 1) and impurity G (epimer 2) = 1.6. Rs = ratio o f the area o f the solvent peak to the area of
Limits: the propanol peak in the chrom atogram obtained
— impurities A (sum of the epimers): m aximum 0.5 per cent; with the reference solution.
— impurity C: maximum 0.4 per cent; Limits:
— impurities E: maximum 0.3 p er cent; — acetone: maximum 1.0 per cent m/m;
— impurities B, F, H: for each impurity, m axim um — methanoh maximum 0.5 per cent m/m;
0.1 per cent; — impurity D: maximum 0.4 per cent m/m.
— impurity G: for each epimer, m axim um 0.1 per cent; H e a v y m e ta ls (2.4.8)
— unspecified impurities: for each impurity, maximum
M axim um 10 ppm .
0.05 per cent;
— total: m axim um 1.0 per cent;
Solvent methanol R.
— reporting threshold: 0.03 per cent; disregard any peak due 1.0 g complies with test H . Prepare the reference solution
to im purity D in the chrom atogram obtained with using 1 m l. o f lead standard solution (10 ppm Pb) R.
reference solution (d). W a te r (2.5.12)
Impurity D , acetone and methanol M aximum 2.0 per cent, determ ined on 0.500 g.
Gas chrom atography (2.2.25). B a c te ria l e n d o to x in s (2.6.14)
Internal standard solution Dissolve 0.5 m l. of propanol R in Less than 0.17 IU/mg, if intended for use in the m anufacture
water R and dilute to 1000 m l. with the same solvent. o f parenteral preparations without a further appropriate
Test solution Dissolve 0.200 g o f the substance to be procedure for the removal o f bacterial endotoxins.
examined in water R, add 2.0 m l. o f the internal standard A SSA Y
solution and dilute to 10.0 m L with water R. Dissolve 0.100 g in 30 m L o f methanol R and add 5 m L of
Reference solution Dissolve 2.0 m l. o f acetone R, 0.5 m L o f water R. A dd 0.1 M hydrochloric acid to a p H o f about 3.0.
methanol R and 0.5 m L of mesityl oxide R (impurity D ) in Carry out a potentiom etric titration (2.2.20), using 0.1 M
water R and dilute to 1000 m L with the same solvent sodium hydroxide. T hree jum ps of potential are observed.
T o 2.0 m L o f this solution add 2.0 m L o f the internal Read the volume added between the 1st and the 3rd point of
standard solution and dilute to 10.0 m L with water R. This inflexion.
solution contains 316 jig o f acetone, 79 jig of m ethanol and 1 m l. o f 0.1 M. sodium hydroxide is equivalent to 19.02 mg
86 ng o f im purity D p er millilitre. o f C i 6H 25N 2N a 0 5S.
Column:
STO RA G E
— material: fused silica;
In an airtight container, at a tem perature o f 2 °C to 8 °C.
— size. I = 30 m , 0 = 0.53 mm;
I f the substance is sterile, store in a sterile, airtight, tam per
— stationary phase: macrogol 20 000 R (film thickness
proof container.
1.0 jim).
Carrier gas helium for chromatography R. IM P U R IT IE S
Flow rate 9 mL/min. Specified impurities A, B, C, D , E, F , G, H
Temperature:
H02CV ^ s
*'« H
H NH2 o
Tbne Temperature
and epimer at S
(min) CC)
Column 0 - 23 50
A. (Z)-7- [(i&S)- [(2i?)-2-amino-2-carboxyethyl] sulfinyl] -2-
2.5 - 5 50 -> 70 [ [[(15)-2,2-dimethylcydopropyI] carbonyl] amino] hept-2-enoic
5-5.5 70 add,
Detector 220
( h3
c
S o lu b ility
Slightly soluble in water, freely soluble in m ethanol and in
h 3c c' h 3
methylene chloride.
0
ID E N T IF IC A T IO N
D . 4-methylpent-3-en-2-one (mesityl oxide), A. Infrared absorption spectrophotometry (2.2.24).
H02C>^ \ S^ S^ ^ ^ V^ C 0 2H Comparison cUazaprU CRS.
H NH2 o
B. Specific optical rotation (see Tests).
TESTS
E. 7-[[(2i?)-2-amino-2-carboxyethyl] sulfanyl] - 2-oxoheptanoic Sp ecific o p tica l ro ta tio n (2.2.7)
add, — 383 to — 399 (anhydrous substance).
HOjC COjH Dissolve 0.200 g in 0.067 M phosphate buffer solution
pH 7.0 R, with the aid o f ultrasound if necessary, and dilute
to 50.0 m L with the same buffer solution. Carry o u t the
d eterm ination at 365 nm .
I m p u rity A
Thin-layer chromatography (2.2.27).
Test solution Dissolve 0.20 g o f the substance to be examined
F. (2)-7-[[(2i?)-2-ainino-2-carboxyethyi] sulfanyl] -2-[(2,3- in methanol R and dilute to 5.0 m L with the same solvent.
dimethyIbut-3-enoyl)amino]hept-2-enoic ad d ,
Reference solution (a) Dissolve 2 mg of cilazapril
impurity A CRS in methanol R and dilute to 50.0 mT. with the
HO2C
same solvent
Reference solution (b) Dissolve 5 m g of cilazapril
impurity A CRS and 5 m g of the substance to be examined in
and epimer at C*
methanol R and dilute to 10.0 m L with the same solvent.
Plate TLC silica gel plate R.
G. (£)-(2&S)-7-[[(2i?)-2-araino-2-carboxyethyl]sulfanyI]-2-
[ [[(15)-2,2-dimethylcydopropyI] carbonyl] amino]hept-3-enoic Mobile phase glacial acetic add. R, water R, hexane R,
add, methanol R, ethyl acetate R (5:5:15:15:60 V/VfV/V/V).
Application 5 pL.
h 2n ^ c o 2h
Development Over a p ath o f 10 cm.
Drying In a current of cold air for 10 m in .
Detection Spray with a freshly prepared mixture o f 1 volume
o f potassium iodobismuthate solution R and 10 volumes o f dilute
H . (Z )-l- [(2-aminoethyI) sulfanyl]-2- [[ [( 1S)-2,2- acetic acid R and then with dilute hydrogen peroxide solution R.
dimethylcyclopropyi] carbonyl] amino] hept- 2 -enoic ad d . System suitability: reference solution (b):
PhEur
— the chromatogram shows 2 clearly separated spots.
Lanit.
— impurity A: any spot due to impurity A is not more
intense than the corresponding spot in the chromatogram
★*★ obtained with reference solution (a) (0.1 per cent).
★ ★
Cilazapril ★ ★ R e la te d su b sta n c e s
***** Liquid chromatography (2.2.29).
Test solution Dissolve 25.0 mg of the substance to be
examined in the mobile phase and dilute to 50.0 mT. with
, HjO
the mobile phase.
Reference solution (a) Dilute 1.0 m L o f the test solution to
50.0 m L with the mobile phase. Dilute 5.0 m L o f this
solution to 20.0 m L with the mobile phase.
C22H3xN305>H20 435.5 92077-78-6
Reference solution (b) Dissolve 5.0 mg of cUazaprU
A ctio n a n d u se impurity D CRS in the test solution and dilute to 10.0 m L
Angiotensin converting enzyme inhibitor. with the test solution.
Column:
PhEtr_________________________________________________________ — sizer. I = 0.25 m, 0 = 4.6 mm;
D E F IN IT IO N — stationary phase: octadecylsOyl silica gel for chromatography R
(15j95)-9-[[(15)-l -(Ethoxycarbonyl) -3-phenylpropyI] amino] - (5 pm).
10-oxooctahydro- 6//-pyridazino [1,2-a] [ l, 2]diazepine-l- Mobile phase Mix 10 volumes o f triethylamine R and
carboxylic a d d monohydrate. 750 volumes o f water R, adjust to p H 2.30 with phosphoric
C o n te n t add R, and add 200 volumes o f tetrahydrofuran R.
98.5 p er cent to 101.5 per cent (anhydrous substance). Flow rate 1.0 mL/min.
CHARACTERS Detection Spectrophotometer at 214 nm.
A p p e a ra n c e Injection 20 pL.
White o r almost white, crystalline powder.
1-556 Cimetidine 2016
A SSA Y DEFINITION
Dissolve 0.300 g in 10 m L o f anhydrous ethanol R and add 2-C yano-1-methyl-3- [-2 -[ [(5-methyl-1//-im idazol-4-
50 m L of water R. T itrate w ith 0.1 M sodium hydroxide, yl) methyl] sulfanyl] ethyl] guanidine.
determ ining the end-point potentiometrically (2.2.20). Carry Content
out a blank titration. 98.5 per cent to 101.5 p er cent (dried substance).
1 m L o f 0.1 M sodium hydroxide is equivalent to 41.75 mg of CHARACTERS
C 22H 31N 3O 5.
Appearance
STO R A G E W hite or almost white powder.
Protected from light. Solubility
IM P U R IT IE S Slightly soluble in water, soluble in ethanol (96 p er cent),
Specified impurities A, B, C, D practically insoluble in methylene chloride. It dissolves in
dilute mineral adds.
It shows polymorphism (5.9).
ID E N T IF IC A T IO N
First identification B
Second identification A, C
A. M d tin g point (2.2.14): 139 °C to 144 °C.
A. R = C (C H 3)3, R ' = C 2H 5: 1,1-dimethylethyl (15,95)-9- If necessary, dissolve the substance to be examined in
[[(5)-1 -(ethoxycarbonyi)-3-phenyipropyl] amino] -10- 2-propanol R, evaporate to dryness and determ ine the m dting
oxooctahydro- 6H -pyridazino[l, 2 -a] [l, 2 ]diazepine-l- point again.
carboxylate, B. Infrared absorption spectrophotom etry (2.2.24).
B. R = R ' = H : (l5,95)-9-[[(5)-l-carboxy-3- Comparison cimetidine CRS.
phenylpropyl] am ino]- 10-oxooctahydro- 6 if-pyridazino [ 1,2 - If th e spectra obtained in the solid state show differences,
a ][ l, 2 ]diazepine-l-carboxylic a d d , dissolve die substance to be examined and the reference
C. R = R ' = C 2H 5: ethyl (l5 ,9 5 )-9 -[[(5 )-l- substance separately in 2-propanol R, evaporate to dryness
(ethoxycarbonyl)-3-phenylpropyl] amino] -10-oxooctahydro- and record new spectra using the residues.
6/f-pyridazino [ 1,2 -<z] [ l , 2]diazepine-l-carboxylate, C. Thin-layer chrom atography (2.2.27).
2016 Cimetidine 1-557
Test solution Dissolve 10 mg o f the substance to be examined identification CRS and the chromatogram obtained with
in methanol R and dilute to 10 m L with the same solvent reference solution (c) to identify the peak due to impurity F.
Reference solution Dissolve 10 m g of cimetidine CRS in Relative retention W ith reference to cimetidine (retention
methanol R and dilute to 10 m L with the same solvent. time = about 18 min): impurity G = about 0.2;
Plate TLC silica gel GF 254 plate R. impurity E = about 0.4; impurity D = about 1.5;
impurity C = about 1.6; impurity B = about 2 .0 ;
Mobile phase concentrated ammonia R, methanol R, ethyl
impurity H = about 2.3; impurity F = about 4.6.
acetate R (15:20:65 V/V/V).
System suitability: reference solution (b):
Application 5 |iL.
— resolution: minimum 1.5 between the peaks due to
Development Over 3/4 o f the plate. impurities D and C.
Drying In a current of cold air. Limits:
Detection Expose to iodine vapour until maximum contrast — correction factors: for the calculation o f content, multiply
has been obtained and examine in ultraviolet light at the peak areas o f the following impurities by the
254 nm. corresponding correction factor: impurity C = 2.5;
Results T he principal spot in the chromatogram obtained with impurity D = 3.3; impurity E = 0.7; impurity G = 0.6.
the test solution is similar in position and size to the principal — impurities B, C, D, E, F, G, H: for each impurity, not
spot in the chromatogram obtained with the reference more than the area o f the principal peak in the
solution. chromatogram obtained with reference solution (a)
(0.2 per cent);
TESTS — unspecified impurities, for each impurity, not more than
Appearance o f solution 0.5 times the area o f the principal peak in the
T h e solution is clear (2.2.1) and not more intensely coloured
chromatogram obtained with reference solution (a)
than reference solution Y 5 (2.2.2, Method II).
(0.10 per cent);
Dissolve 3.0 g in 12 m L o f 1 M hydrochloric acid and dilute — total: n o t m ore than 5 times the area o f the principal peak
to 20 m L with water R. in the chromatogram obtained with reference solution (a)
Related substances ( 1.0 per cent);
Liquid chromatography (2.2.29). — disregard limit: 0.25 times the area o f the principal peak in
Test solution Dissolve 20 m g of the substance to be examined the chromatogram obtained with reference solution (a)
in mobile phase A and dilute to 50.0 m L with mobile (0.05 per cent).
phase A. H eav y m e ta ls (2.4.8)
Reference solution (a) Dilute 1.0 m L of the test solution to M aximum 20 ppm .
100.0 m L with mobile phase A. Dilute 2.0 m L of this 1.0 g complies w ith test C. Prepare the reference solution
solution to 10.0 m L with mobile phase A using 2 m L o f lead standard solution (10 ppm Pb) R.
Reference solution (b) Dissolve the contents o f a vial o f L oss o n d ry in g (2.2.32)
cimetidine for system suitability CRS (containing impurities 6 , M aximum 0.5 per cent, determined on 1.000 g by drying in
C , D , E, G and H ) in 1.0 m L o f mobile phase A. an oven at 105 °C.
Reference solution (c) Dissolve 4 mg of cimetidine for peak S u lfa te d a s h (2.4.14)
identification CRS (containing impurity F) in mobile phase A M aximum 0.2 per cent, determined on 1.0 g.
and dilute to 10.0 m L with mobile phase A.
A SSAY
Column:
Dissolve 0.200 g in 60 m L of anhydrous acetic acid R T itrate
— size: I = 0.25 m , 0 = 4.6 mm;
with 0.1 M perchloric acid determ inin g the end point
— stationary phase: end-capped octadecylsüyl silica gel for
potentiometrically (2.2.20).
chromatography R (5 Jim).
1 m L o f 0.1 M perchloric acid is equivalent to 25.23 mg of
Mobile phase A M ix 0.4 volumes of dieihylamine R and
c 10H16N6s
780 volumes o f a 1.1 g/L solution of sodium
hexanesulfonaxe R j adjust to p H 2.8 with phosphoric add R; STORAGE
add 250 volumes of methanol R2; Protected from light.
Mobile phase B methanol R2; IM P U R IT IE S
Specified impurities B, C , D , E, F , G, H
Time Mobile phase A Mobile phase B Other detectable impurities (the following substances would, if
(min) (per cent V/V) (per cent V/V) present at a sufficient level, be detected by one or other o f
0 -6 0 100 0 the tests in the monograph. They are limited by the general
6 0 -6 5 100*90 0 * 10 acceptance criterion for other/unspecified impurities and/or
65-120 90 10
by the general monograph Substances for pharmaceutical use
(2034). It is therefore n o t necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Flow rate 1.1 mL/min. Control of impurities in substances for pharmaceutical use): A , I,
Detection Spectrophotom eter at 220 nm.
Irgection 50 |il.
Identification of impurities Use the chromatogram supplied
with cimetidine for system suitability CRS and the
chromatogram obtained with reference solution (b) to
identify the peaks due to impurities B, C, D , E, G and H ;
use the chromatogram supplied with cimetidine for peak
1-558 Cimetidine Hydrochloride 2016
★*★
★ ★
Cimetidine Hydrochloride ★ ★
(Ph. Ever, monograph 1500) *****
R1
I
N
HN
C 10H 17C1N6S 288.8 70059-30-2
H
H3C Action and use
H istamine H 2 receptor antagonist; treatm ent o f peptic
B. R I = C N , R2 = 0 -C H 3, X = S: methyl 3-cyano- 1-[ 2- ulceration.
[ [(5-methyl-1H-imidazol-4- Preparations
yl) methyl] sulfanyl] ethyl] carbamimidate, Cimetidine Injection
C. R I = C O -N H * R 2 = N H -C H 3, X = S:
1- [(m ethylamino) [[2- [ [(5-methyl-l //-im idazol-4- PhEur.
y 1) methyl] sulfanyl] ethyl] amino] methylidene] urea, D E F IN IT IO N
D . R I = H , R 2 = N H -C H 3, X = S: l-m ethyl-3-[2- 2-C yano-1-methyl-3- [-2- [[(5-methyi- 1if-im idazol-4-
[ [(5-methyl-1H-imidazol-4- yQmethyl] sulfanyl] ethyl] guanidine hydrochloride.
yl)methyl] sulfanyl] ethyl] guanidine,
Content
E. R I = C N , R2 = N H -C H 3, X = SO: 2-cyano - 1-methyl- 98.5 p e r cent to 101.5 per cent (dried substance).
3 [2- [ [(5-methyl- 1/i-im idazol-4-
CHARACTERS
yl) methyl] sulfinyl] ethyl] guanidine,
Appearance
W hite or alm ost white, crystalline powder.
S o lu b ility
Freely soluble in water, sparingly soluble in anhydrous
ethanol.
ID E N T IF IC A T IO N
First identification B, D
F. 2-cyano-1,3-bis [2- [ [(5-methyl- lH-imidazol-4- Second identffication A , C, D
yl) methyl] sulfanyl] ethyl] guanidine, A. Ultraviolet and visible absorption spectrophotom etry
(2.2.25).
, cn
N Test solution Dissolve 70 m g in 0 . 2 M sulfuric acid and dilute
h3C. . JL .CHj
N N
to 100.0 m L with the same ad d . D ilute 2.0 m L o f this
H H solution to 100.0 m L with 0 . 2 M sulfuric add.
Specific absorbance at the absorption maximum at 218 nm
G. 2-cyano-1 , 3 -dimethylguanidine, 650 to 705.
B. Infrared absorption spectrophotom etry (2.2.24).
Comparison dmeudme hydrochloride CRS.
C . Thin-layer chromatography (2.2.27).
Test solution Dissolve 10 m g o f the substance to be examined
in methanol R and dilute to 10 m L with the same solvent.
Reference solution Dissolve 10 mg o f dmetidine
hydrochloride CRS in methanol R and dilute to 10 m L w ith the
H . 1,1 '-(disulfanediyldiethylene)bis(2-cyano-3- same solvent.
methylguanidine), Plate TLC silica gel GF2 5 4 plate R.
Mobile phase concentrated ammonia R, methanol R, ethyl
/^ n acetate R (15:20:65 VIVIV).
HN
R1 Application 5 pL.
R2 Development Over 3/4 o f the plate.
Drying In a current o f cold air
I. R I = O H , R 2 = C 2H 5: (5-ethyl-1//-im idazol-4- Detection Expose to iodine vapour until maximum contrast
yl)methanol, has been obtained and examine in ultraviolet light at
J. R I = S-C H z-C H z-N H * R2 = C H 3: 2-[[(5-m ethyl-lH - 254 nm .
imidazol-4-yl)methyl] sulfanyl] ethanamine. Results T he principal spot in the chromatogram obtained with
PhEur the test solution is similar in position and size to the principal
spot in the chrom atogram obtained with the reference
solution.
2016 Cimetidine Hydrochloride 1-559
***
★ ★
Cinchocaine Hydrochloride ★ ★
(PL Eur. monograph 1088) * * * * *
R e la te d su b s ta n c e s IM P U R IT IE S
Gas chrom atography (2.2.28).
ch 3
Internal standard solution Dissolve 1.0 g o f camphor R in
heptane R and dilute to 200 m L with the same solvent.
Test solution (a) Dissolve 2.5 g o f die substance to be
examined in heptane R and dilute to 25.0 m L with the same
solvent.
Test solution (b) Dissolve 2.5 g o f the substance to be
examined in heptane R, add 5.0 m L o f the internal standard A. l-methyl-4-(l-methylethyl)-7-oxabicyclo[2.2. l]heptane
solution and dilute to 25.0 m L with heptane R. (1,4-cineole).
Reference solution (a) T o 2.0 m L o f test solution (a) add ___________________________________________________________PhEur
20.0 m L of the internal standard solution and dilute to
100.0 m L w ith heptane R.
Reference solution (b) Dissolve 50 m g o f 1,4-cineole R and
50 m g o f the substance to be examined in heptane R and
dilute to 50.0 m L w ith the same solvent. Cinnamic Acid
Column:
— sizer. I = 30 m , 0 = 0.25 m m ,
— stationary phase: macrogol 20 000 R (film thickness
0.25 jam).
Carrier gas helium for chromatography R.
Linear velocity 45 cm/s. G>H80 2 148.2 621-82-9
SpUt-ratio 1:70.
A c tio n a n d u se
Temperature: Antimicrobial preservative; excipient.
lim e Temperature D E F IN IT IO N
(min) CO Cinnamic A d d is (£)-3-phenylprop-2-enoic acid. It contains
Column 0 - 10 50 n o t less than 99.0% and not more than 100.5% of CgH aO ^
10-35 5 0 * 100 calculated with reference to the dried substance.
35 -4 5 100*200
45 -5 5 200
C H A R A C T E R IS T IC S
Injection port 220 Colourless crystals.
Very slighdy soluble in water, freely soluble in ethanol (96%)',
Detector 250
soluble in ether.
ID E N T IF IC A T IO N
Detection Flam e ionisation. A. T he infrared absorption spectrum, Appendix II A, is
Iiyecdon 1 |iL. concordant with the reference spectrum of cinnamic acid
System suitability, reference solution (b): (RS 062).
— resolution: m inim um 10 between the peaks due to B. T h e light absorption, Appendix II B, in the range 230 to
im purity A and to cineole. 350 nm o f a 0.0010% w/v solution in 0.1m sodium hydroxide
Limits: exhibits a maximum only at 267 nm . T he absorbance at
— total: calculate the ratio (R) o f the area of the peak due to 267 nm is about 1.4.
cineole to the area of the peak due to the internal
TESTS
standard from the chrom atogram obtained with reference
M e ltin g p o in t
solution (a); from the chrom atogram obtained with test
132° to 134°, Appendix V A.
solution (b), calculate the ratio o f the sum of the areas of
any peaks, apart from the principal peak and the peak E th a n o l-in s o h ib le m a tte r
due to the internal standard, to the area o f the peak due A 10% w/v solution in ethanol (96%) is clear, Appendix IV A.
to internal standard: this ratio is no t greater than R~ R e la te d su b s ta n c e s
(2 per cent), Carry out the m ethod for thin-layer chromatography,
— disregard limit. 0.025 times the area o f the principal peak A ppendix m A, using the following solutions in methanol.
in the chrom atogram obtained with reference solution (a) (1) 5.0% w/v o f the substance being examined.
(0.05 per cent).
(2) 0.025% w/v of the substance being examined.
R e sid u e o n e v a p o ra tio n
CHROMATOGRAPHIC CONDITIONS
M axim um 0.1 per c e n t
(a) Use as the coating silica gel GF2S4.
T o 2.0 g add 5 m L o f water R, evaporate to dryness on a
water-bath and dry at 100-105 °C for 1 h. T h e residue (b) Use the mobile phase as described below.
weighs a m axim um o f 2 mg. (c) Apply 5 *iL o f each solution.
STORAGE (d) Develop die plate to 15 cm.
In an airtight container, protected from lig h t (e) After removal o f the plate, dry in air and examine under
ultraviolet light (254 nm).
MOBILE PHASE
10 volumes o f glacial acetic acid and 90 volumes o f toluene.
2016 Cinnarizine 1-563
Dissolve 1.0 g in anhydrous ethanol R and dilute to 10.0 m L — disregard limit. 0.5 times the area of the principal peak in
w ith the same solvent. the chromatogram obtained w ith reference solution (a)
R e la te d su b s ta n c e s (0.05 p er cent).
L iquid chromatography (2.2.29). C h lo rid e s (2.4.4)
Test solution Dissolve 0.125 g o f the substance to be M axim um 350 ppm.
exam ined in a mixture o f equal volumes o f acetomtrUe R and T o 0.190 g add 20 m l. o f water R and treat in an ultrasonic
water R and dilute to 50 m L with the same mixture of bath for 8 min. Filter. 15 mL of the filtrate complies with the
solvents. test.
Reference solution (a) Dilute 1.0 m L of the test solution to W a te r (2.5.12)
100.0 m L with a mixture of equal volumes o f acetomtrUe R M aximum 0.5 per cent, determined on 1.000 g.
and water R. D ilute 1.0 m L o f this solution to 10.0 m L with S u lfa te d a s h (2.4.14)
a m ixture of equal volumes of acetomtrUe R and water R.
M aximum 0.1 p er cent, determined on 1.0 g.
Reference solution (b) Dissolve the contents o f a vial o f
A SSAY
ciprofibrate for system suitability CRS in 2.0 m L o f a mixture of
equal volumes o f acetomtrUe R and water R. Dissolve 0.250 g in a mixture o f 20 m L of water R and
40 m L o f anhydrous ethanol R. T itrate with 0.1 M sodium
Column'.
hydroxide, d eterm ining the end-point potentiometrically
— size: I = 0.15 m , 0 = 4.6 mm,
(2.2.20).
— stationary phase. octylsUyl silica gel for chromatography R
(5 pm). 1 m L o f 0.1 M sodium hydroxide is equivalent to 28.92 m g
of C 13H 14CI2O 3.
Mobile phase:
— mobile phase A: 1.36 g/L solution of potassium dihydrogen STORAGE
phosphate R adjusted to p H 2.2 with phosphoric acid R, In an airtight container, protected from light
— mobile phase B: acetomtrUe R,
IM P U R IT IE S
Specified impurities A, B, C, D , E.
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0 -3 0 7 5 -» 3 0 2 5 -» 7 0 f Y ° x COlH
3 0 -40 30
h 1c » J v ) h >c
70
40-42 3 0 -» 7 5 7 0 -» 2 5
A. 2-(4-ethenylphenoxy)-2-methylpropanoic ad d ,
★ ★ Limit.
Ciprofloxacin ★ ★ — impurity A: any spot due to impurity A is n o t more
★ intense than the principal spot in the chrom atogram
(Ph. Eur. monograph 1089)
obtained with the reference solution (0.2 p er cent).
R elate d su b s ta n c e s
Liquid chromatography (2.2.29).
Test solution T o 25.0 mg of the substance to be examined add
0.2 m L o f dilute phosphoric acid R and dilute to 50.0 m L with
C02H the mobile phase. T reat in an ultrasonic bath until a clear
solution is obtained.
Reference solution (a) Dilute 1.0 m L o f the test solution to
C 17H 18FN 3O 3 331.4 85721-33-1 100.0 m L w ith the mobile phase. Dilute 1.0 m L of this
solution to 5.0 m L with the mobile phase.
Action and use
Reference solution (b) Dissolve 2.5 m g of dprofloxadn
Fluoroquinolone antibacterial.
hydrochloride for peak identification CRS (containing
Preparations impurities B, C , D and E) in the mobile phase and dilute to
Ciprofloxacin Infusion 5.0 m L with the mobile phase.
Ciprofloxacin Eye Drops Column:
— size: I = 0.25 m , 0 = 4.6 mm;
PhEtr__________________________
— stationary phase: base-deactivated end-capped octadecylsifyl
D E F IN IT IO N silica gel for chromatography R (5 ¡mi);
1-Cyclopropyl-6-fluoro-4-oxo-7 - (piperazin-1-yl)-1,4- — temperature:: 40 °C.
dihydroquinoline-3-carboxylic ad d . Mobile phase Mix 13 volumes o f acetonitrUe R and 87 volumes
Content o f a 2.45 g/L solution of phosphoric add R, previously
99.0 per cent to 101.0 per cent (dried substance). adjusted to p H 3.0 with triethylamine R.
CHARACTERS Flow rate 1.5 mL/min.
Appearance Detection Spectrophotom eter at 278 nm .
Almost white or pale yellow, crystalline powder, slightly Injection 50 |iL.
hygroscopic. Run time Twice the retention time of ciprofloxacin.
Solubility Identification of impurities Use the chrom atogram supplied
Practically insoluble in water, very slightly soluble in with ciprofloxacin hydrochloride for peak identification CRS and
anhydrous ethanol and in methylene chloride. the chromatogram obtained with reference solution (b) to
ID E N T IF IC A T IO N identify the peaks due to impurities B, C , D and E.
Infrared absorption spectrophotometry (2.2.24). Relative retention W ith reference to ciprofloxacin (retention
Comparison ciprofloxacin CRS. time = about 9 min): impurity E — about 0.4;
impurity B = about 0.6; impurity C = about 0.7;
TESTS impurity D = about 1.2.
Appearance o f solution System suitability: reference solution (b):
T he solution is clear (2.2.1) and not more intensely coloured — resolution: minimum 1.3 between the peaks due to
than reference solution GY 5 (2.2.2, Method II). impurities B and C.
Dissolve 0.25 g in a 10.3 g/L solution o f hydrochloric add R Limits:
and dilute to 20 m L with the same solution. — correction factors: for the calculation o f content, multiply
Impurity A the peak areas o f the following impurities by the
Thin-layer chromatography (2.2.27). corresponding correction factor, impurity B = 0.7;
Test solution Dissolve 50 m g of the substance to be examined impurity C = 0.6; impurity D = 1.4; impurity E = 6.7;
in dilute ammonia R1 and dilute to 5 m L with the same — impurities B, C, D, E: for each impurity, n o t more than
solvent. the area o f the principal peak in the chromatogram
obtained with reference solution (a) (0.2 p er cent);
Reference solution Dissolve 10 m g of ciprqfloxadn
— unspecified impurities: for each impurity, no t more than
impurity A CRS in a mixture o f 0.1 m L of dilute ammonia R1
0.5 times the area of the principal peak in the
and 90 m L o f water R and dilute to 100 m L with water R.
chromatogram obtained with reference solution (a)
Dilute 2 m L o f the solution to 10 m l. with water R.
(0.10 p er cent);
Plate TLC silica gel F2 $ 4 plate R. — total: n o t more than 2.5 times the area of the principal
Mobile phase acetomtrUe R, concentrated ammonia R, peak in the chromatogram obtained with reference
methanol R, methylene chloride R (10:20:40:40 V/V/V/V). solution (a) (0.5 per cent);
Application 5 |iL. — disregard limit: 0.25 times the area of the principal peak in
Development A t the bottom of a chromatographic tank, place the chromatogram obtained with reference solution (a)
an evaporating dish containing 50 m L o f concentrated (0.05 p er cent).
ammonia R. Expose the plate to the ammonia vapour for H eav y m e ta ls (2.4.8)
15 m in in the closed tank. W ithdraw the plate, transfer to a M aximum 20 ppm.
2nd chromatographic tank and develop over 3/4 of the plate. Dissolve 0.5 g in dilute acetic add R and dilute to 30 m L with
Drying In air. the same solvent. A dd 2 m L of water R instead of 2 m L of
Detection Examine in ultraviolet light at 254 nm. buffer solution pH 3.5 R T he filtrate complies with test E.
2016 Ciprofloxacin Hydrochloride 1-567
a HCl , x HzO
X jÇ u
o
C02H
p Y A c tio n a n d u se
Fluoroquinolone antibacterial.
Xxjx o
P r e p a r a tio n
Ciprofloxacin Tablets
PhEur.
B. l-cyclopropyl-4-oxo-7-(piperazin-l-yl)-l,4- D E F IN IT IO N
dihydroquinoline-3-carboxylic a d d (desfluoro compound),
1-Cydopropyl-6-fluoro-4-oxo-7-(piperazin - 1-yl)-1,4-
dihydroquinoline-3-carboxylic a d d hydrochloride. It contains
H,N'
. Y a variable quantity o f water.
C o n te n t
98.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
o
A p p e a ra n c e
C. 7-[(2-aminoethyI)amino] -1 -cyclopropyl-6-fluoro-4-oxo- Pale yellow, crystalline, slightly hygroscopic powder.
134-dihydroquinoline-3-carboxylic a d d (ethyienediamine S o lu b ility
com pound), Soluble in water, slightly soluble in methanol, very slightly
soluble in anhydrous ethanol, practically insoluble in acetone,
in ethyl acetate and in methylene chloride.
ID E N T IF IC A T IO N
CO2H A. Infrared absorption spectrophotometry (2.2.24).
Comparison ciprofloxacin hydrochloride CRS.
B. 0.1 g gives reaction (b) o f chlorides (2.3.1).
D . 7-chloro- l-cydopropyl-4-oxo-6-(piperazin-1-yl)-1,4-
dihydroquinoline-3-carboxylic ad d ,
1-568 Ciprofloxacin Hydrochloride 2016
>*★
Cisplatin ★ ★
★ ★
(Ph. Eur. monograph 0599) *****
COjH
c iN x nh3
Pt
ay x nh3
A. 7-chloro-1-cydopropyl-6-fluoro-4-oxo-1,4-
dihydroquinoline-3-carboxylic a d d (fluoroquinolonic
add), PtCl2(NH 3)2 300.0 15663-27-1
PhEur.
DEFINITION
B. 1-cyclopropyl-4-oxo-7-(piperazin - 1-yl) - 1,4- dy-Diamminedichloroplatinum(II).
dihydroquinoline-3-carboxylic a d d (desfluoro com pound), Content
97.0 per cent to 102.0 per cent.
CHARACTERS
Appearance
Yellow powder, o r yellow or orange-yellow crystals.
c o 2h
Solubility
Slightly soluble in water, sparingly soluble in
dimethylformamide, practically insoluble in ethanol
(96 per cent).
C . 7-[(2 -aminoethyI)amino]-1 -cyclopropyl-6-fluoro-4-oxo-
l,4-dihydroquinoline-3-carboxyIic a d d (ethylenediamine Cany out identification test B, the tests (except that for silver)
com pound), and the assay protected from light.
IDENTIFICATION
First identification A , B.
Second identification B, C.
A. Infrared absorption spectrophotometry (2.2.24).
COjH Comparison cisplatin CRS.
B. Thin-layer chromatography (2.2.27).
Test solution Dilute 1 m L of solution S2 (see Tests) to 10 m L
with dimethylformamide R.
D . 7-chloro-1-cyclopropyl-4-oxo-6-(piperazin-1-yl)-1,4-
dihydroquinoline-3-carboxyüc ad d , Reference solution Dissolve 10 mg o f cisplatin CRS in 5 m L of
dimethylformamide R
Plate cellulose for chromatography R1 as the coating substance.
Y Pretreatment Activate the plate by hearing at 150 °C for 1 h.
:x)Çi o
Mobile phase acetone R , dimethylformamide R (10:90 VIV).
Application 2 |iL.
Development Over 2/3 of the plate.
Drying In air.
E. l-cyclopropyl-6-fhioro-7-(piperazin-l-yl)quinolin-4(lif)- Detection Spray with a 50 g/L solution o f stannous chloride R
one (decarboxylated compound), in a mixture o f equal volumes of dilute hydrochloric acid R and
water R. Examine after 1 h.
Results T h e principal spot in the chromatogram obtained with
the test solution is similar in position, colour and size to the
principal spot in the chromatogram obtained with the
reference solution.
C. Add 50 mg to 2 m L o f dilute sodium hydroxide solution R in
a glass dish. Evaporate to dryness. Dissolve the residue in a
mixture o f 0.5 m L of nitric add R and 1.5 m L of hydrochloric
F. 1-cyclopropyl-6-hydroxy-4-oxo-7-(piperazin-1-yl)- 1,4- add R. Evaporate to dryness. T he residue is orange. Dissolve
dihydroquinoline-3-carboxylic add. the residue in 0.5 m L of water R and add 0.5 m L of
PhEir ammonium chloride solution R. A yellow, crystalline predpitate
is formed.
1-570 Cisplatin 2016
A p p e a ra n c e o f so lu tio n S I
Limits:
— impurity A: not m ore than the area of the corresponding
Solution S I is clear (2.2. 1) and not m ore intensely coloured
peak in the chromatogram obtained with reference
than reference solution GY 5 (2.2.2, Method II).
solution (d) (2.0 per cent);
A p p e a ra n c e o f so lu tio n S2 — impurity B: not m ore than the area o f the corresponding
Solution S2 is clear (2.2.1). peak in the chromatogram obtained with reference
p H (2.2.3) solution (d) ( 1.0 per cent);
4.5 to 6.0 for solution S I, m easured immediately after — unspecified impurities: for each impurity, not more than
preparation. 0.5 times the area o f the peak due to cisplatin in the
R e la te d su b s ta n c e s chromatogram obtained with reference solution (d)
Liquid chromatography (2.2.29). Carry out the test protected ( 0.10 per cent);
from light. Do not heat or sonicate any platinum-containing — sum of impurities other than A and B: not more than
solution. All solutions are to be used within 4 h. 2.5 times the area o f the peak due to cisplatin in the
chromatogram obtained with reference solution (d)
Test solution Dissolve 25.0 m g o f the substance to be
(0.5 per cent);
examined in a 9.0 g/L solution of sodium chloride R and dilute
— disregard Umit. the area o f the peak due to cisplatin in the
to 25.0 m L with the same solution.
chromatogram obtained with reference solution (e)
Reference solution (a) Dissolve 25.0 mg of cisplatin CRS in a (0.05 per cent). Disregard any peak due to the cisplatin
9.0 g/L solution o f sodium chloride R and dilute to 25.0 m L aquo complex.
with the same solution.
S ilv e r
Reference solution (b) Dissolve 5.0 m g o f cisplatin M aximum 250 ppm.
impurity A CRS in a 9.0 g/L solution o f sodium chloride R and
Atomic absorption spectrometry (2.2.23, Method I).
dilute to 50.0 m L w ith the same solution.
Reference solution (c) Dissolve 5.6 mg o f cisplatin Test solution Dissolve 0.100 g in 15 m L of nitric acid R,
impurity B CRS in a 9.0 g/L solution o f sodium chloride R and heating to 80 °C. Cool and dilute to 25.0 m L with water R.
dilute to 100.0 m L with the same solution. Reference solutions T o suitable volumes (10 m L to 30 m L) of
Reference solution (d) M ix 0.05 m L of the test solution with silver standard solution (5 ppm Ag) R add 50 m L o f nitric
5.0 m L of reference solution (b) and 5.0 m L of reference acid R and dilute to 100.0 m L with water R.
solution (c) and dilute to 25.0 m L with a 9.0 g/L solution of Source Silver hollow-cathode lamp, preferably using a
sodium chloride R. transmission band of 0.5 nm .
Reference solution (e) Dilute 5.0 m L of reference solution (d) Wavelenth 328 nm.
to 20.0 m L with a 9.0 g/L solution o f sodium chloride R. Atomisation device Fuel-lean air-acetylene flame.
Blank solution 9.0 g/L solution o f sodium chloride R. Carry out a blank determination.
Column: A SSA Y
— sizer. I = 0.25 m , 0 = 4.0 mm; Liquid chromatography (2.2.29) as described in the test for
— stationary phase: base-deactivated octylsüyl silica gel for related substances with the following modification.
chromatography R (4 pm);
— temperature. 30 °C.
Injection 10 nL of the test solution and reference solution (a).
Calculate the percentage content o f PtC l 2(N H 3)2 from the
Mobile phase Dissolve 1.08 g o f sodium octanesulfonate R,
sum of the areas o f the peaks due to cisplatin and cisplatin
1.70 g of tetrabutylammonium hydrogen sulfate R and 2.72 g of
aquo complex and from the declared content of
potassium dihydrogen phosphate R in waterfor chromatography R
and dilute to 950 m L with the same solvent. Adjust to cisplatin CRS.
p H 5.9 with 1 M sodium hydroxide and dilute to 1000 m L STORAGE
with water for chromatography R. In an airtight container, protected from light.
Flow rate 1.0 mlVmin. IM P U R IT IE S
Detection Spectrophotom eter at 210 nm . Specified impurities A, B
Injection 20 pL o f the test solution, reference solutions (d) Other detectable impurities (the following substances would, if
and (e), and the blank solution. present at a sufficient level, be detected by one or other of
Run time 7 times the retention time o f cisplatin. the tests in the monograph. T hey are limited by the general
T h e displacement peak is the latest eluting peak of the group acceptance criterion for other/unspecified impurities and/or
of injection peaks in the chrom atogram obtained with the by the general monograph Substances for pharmaceutical use
blank solution. (2034). It is therefore n o t necessary to identify these
impurities for demonstration o f compliance. See also 5.10.
Identification of cisplatin aquo complex Use the chromatogram
Control of impurities in substances for pharmaceutical use): C.
supplied with cisplatin CRS and the chromatogram obtained
with reference solution (a) to identify the peak due to
cisplatin aquo complex.
2016 Citalopram Hydrobromide 1-571
C IN x nh3
Test solution Dissolve 50 mg of the substance to be examined
Pt
✓ \ in mobile phase A and dilute to 100.0 m L w ith mobile
H3N CI
phase A.
A. rran$-diamminedichloroplatinum(II) (transplatin), Reference solution (a) Dilute 1.0 m L of the test solution to
100.0 m L with mobile phase A (solution A). Dilute 1.0 mT.
o f solution A to 10.0 m L with mobile phase A.
C IN x NH3"
Pt
Reference solution (b) Dissolve the contents o f a vial of
a y v ci citalopram for system suitability CRS (containing impurities B,
D and G) in 1.0 m L o f solution A.
Column:
B. am m inetrichloroplatm ate(-)j
— size: I = 0.25 m , 0 = 4.6 m m ;
— stationary phase, end-capped octadecylsüyl sUica gel for
a \ , aT chromatography R (4 |im);
Pt — temperature: 40 °C.
a y yci
Mobile phase:
— mobile phase A: dissolve 1.58 g of ammonium formate R in
C. tetrachloroplatinate( 2-). 500 m L of a mixture of 4 volumes o f acetomtrUe R,
: _______________________________ ______ _ PhEur 32 volumes o f methanol R and 64 volumes o f water R;
— mobile phase B: dissolve 1.58 g of ammonium formate R in
500 m L of a mixture of 32 volumes o f water R and
68 volumes o f acetonitrUe R;
H e a v y m e ta ls (2.4.8)
M axim um 20 ppm .
Dissolve 0.5 g in ethanol (96 per cent) R and dilute to 20 mT. and enantiomer
w ith the same solvent. 12 m L o f the solution complies with
test B. Prepare the reference solution using lead standard
solution (0.5 ppm Pb) obtained by diluting lead standard
solution (100 ppm Pb) R w ith ethanol (96 per cent) R. Filter
the solutions through a m em brane filter (nominal pore size D . R l = C N , R2 = H: (1 RS) -1 - (4-fluorophenyl)-1- [3-
0.45 pm). (methylamino)propyl] -1,3-dihydroisobenzofuran-5-
carbonitrile,
L o ss o n d ry in g (2.2.32)
M axim um 0.5 per cent, determ ined on 1.000 g by drying in F. R l = Br, R2 = C H 3: 3-[(lÄ S )-5-brom o-l-
an oven at 105 °C for 4 h. (4-fluorophenyl)-1,3-dihydro isobenzofuran-1-yl] -N,N-
dim ethylpropan- 1-amine,
S u lfa te d a s h (2.4.14)
M axim um 0.1 per cent, determ ined on 1.0 g in a platinum G. R l = C O -[C H 2] 3-N (C H 3)2, R2 = C H 3:
crucible. 4-(dimethylamino) -1 - [(1RS) -1 - [3-(dimethylamino)propyl] -1 -
(4-fluorophenyl)-1,3-dihydroisobenzofuran-5 -yl] b utan-1-one.
A SSA Y
__________________________________________________________ PhEur
Dissolve 0.300 g in 50 m L o f ethanol (96 per cent) R and add
0.5 m L o f 0.1 M hydrochloric acid Carry out a potentiom etric
titration (2.2.20), using 0.1 M sodium hydroxide. Read the
volum e added between the 2 points o f inflexion.
1 m L o i 0.1 M sodium hydroxide is equivalent to 40.53 mg o f Citalopram Hydrochloride ★★*★★
★ ★
C 2oH 22B rFN 20 .
(Ph. Eur. monograph 2203) *****
IM P U R IT IE S
Specified impurities B, D, G
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other o f
the tests in the monograph. T hey are limited by the general Ha
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). I t is therefore not necessary to identify these
impurities for dem onstration o f compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): A , C, C 20H 22CIFN 2O 360.9 85118-27-0
E ,F .
A c tio n a n d u se
Selective serotonin reuptake inhibitor; antidepressant.
P re p a r a tio n s
Citalopram Oral Drops
.c h 3 and enantiomer PhEur.
N
CH3 D E F IN IT IO N
(1 RS ) - 1-[3-(Dimethylamino)propyl]- l-(4-fluorophenyl)-1,3-
dihydroisobenzofuran-5-carbonitrile hydrochloride.
C o n te n t
A. R = C O -N H 2, X = H 2: (l.ftS )-l-[3-
99.0 per cent to 101.5 per cent (dried substance).
(dimethylamino)propyl] -1 -(4-fluorophenyl)-1,3-
dihydroisobenzofuran-5-carboxamide, CHARACTERS
C. R = C N , X = O: (3&S)-6-cyano-3-[3- A p p e a ra n c e
(dimethylamino)propyl]-3-(4-fluorophenyI)isobenzofuran- W hite or almost white, crystalline powder.
1 (3H)-one, S o lu b ility
E. R = Cl, X = H 2: 3-[( 1RS) -5-chloro-1- (4-fluorophenyl) - Very soluble in water, freely soluble in anhydrous ethanol.
1,3-dihydroisobenzofuran-l -yl] -N ^-d im eth y lp ro p an -1- ID E N T IF IC A T IO N
am ine, A. Optical rotation (see Tests).
B. Infrared absorption spectrophotometry (2.2.24).
Comparison citalopram hydrochloride CRS.
C. It gives reaction (a) of chlorides (2.3.1).
its epimer at C*
ch3
and their enantiomers TESTS
N
1 S o lu tio n S
ch3
Dissolve 1.0 g in methanol R and dilute to 20 m L with the
same solvent.
A p p e a ra n c e o f so lu tio n
B. l-[3-(dim ethylam ino)propyl]-l-(4-fluorophenyl)-3- Solution S, examined immediately after preparation, is clear
hydroxy-133dihydroisobenzofuran-5-carbonitrile, (2.2.1) and not m ore intensely coloured than reference
solution Y 6 (2.2.2, Method IT).
2016 Citalopram Hydrochloride 1-573
2 -2 5 100->40 0*60
2 5 -3 0 40 60
CH3 and enantiomer
B. Infrared absorption spectrophotometry (2.2.24). Reference solution (c) Dilute 1.0 mL of reference solution (b)
Comparison cladribine CRS. to 10.0 mL with the solvent mixture.
If the spectra obtained in the solid state show differences, Reference solution (d) Dissolve 1.0 mg of cladribine
dissolve the substance to be examined in the minimum impurity C CRS in reference solution (b) and dilute to
volume of methanol R and evaporate to dryness. Dry the 25.0 mL with the same solution.
precipitate at 100 °C for 2 h and record a new spectrum Reference solution (e) Dilute 5.0 mL of reference solution (c)
using the residue. to 10.0 mL with the solvent mixture.
TESTS Reference solution (f) Dissolve 3 mg of cladribine for peak
Appearance of solution identification CRS (containing impurities A, B, C and D) in
The solution is clear (2.2.1) and colourless (2.2.2, 2 mL of the solvent mixture.
Method II). Column:
Disperse 0.15 g in water R, dilute to 50 mL with the same — size:. I = 0.25 m, 0 = 4.6 mm;
solvent and sonicate until dissolution is complete. — stationary phase, base-deactivated octylsüyl silica gelfor
chromatography R (5 pm).
Specific optical rotation (2.2.7)
Mobile phase:
—21.0 to —27.0 (anhydrous substance).
— mobile phase A: waterfor chromatography R;
Dissolve 0.25 g in dimethyl sulfoxide R and dilute to 25.0 mL — mobile phase B: acetonitrile for chromatography R;
with the same solvent. — mobile phase C: 50 g/L solution of phosphoric acid R in
Im purity E waterfor chromatography R;
Thin-layer chromatography (2.2.27).
Test solution Dissolve 40.0 mg of the substance to be Time Mobile phase A Mobile phase B Mobile phase C
examined in dimethyJformamide R and dilute to 2.0 mL with (min) (per cent V/V) (per cent V/V) (per cent V/V)
the same solvent. 0 -1 0 80*70 10 * 2 0 10
Reference solution (a) Dissolve 5.0 mg of 2-deoxy-n-ribose R 10-25 70*20 20*70 10
(impurity E) in dimethylformamide R and dilute to 25.0 mL
25-30 20 70 10
with the same solvent. Dilute 3.0 mL of this solution to
10.0 mL with dimethylformamide R.
Reference solution (b) Dissolve 10.0 mg of 2-deoxy-u-ribose R Flow rate 0.8 mlVmin.
(impurity E) in dimethylformamide R and dilute to 5.0 mL Detection Spectrophotometer at 252 nm.
with the same solvent. Mix 9 volumes of this solution with
Injection 20 |iL of test solution (a) and reference
1 volume of the test solution.
solutions (c), (d), (e) and (f).
Plate TLC silica gel F 254 plate R.
Identification of impurities Use the chromatogram supplied
Mobile phase concentrated ammonia R, ethanol (96 per cent) R, with cladribine for peak identification CRS and the
ethyl acetate R (20:40:40 VIVIV). chromatogram obtained with reference solution (f) to identify
Application 5 nL as bands of 10 mm; thoroughly dry the the peaks due to impurities A, B, C and D.
points of application in a current of warm air. Relative retention With reference to cladribine (retention
Development Over 2/3 of the plate. time = about 10 min): impurity A = about 0.33;
Drying In air, then heat at 45 °C for 10 min. impurity B = about 0.44; impurity C = about 0.73;
Detection Spray with a solution containing 0.5 g of thymol R impurity D = about 0.92.
in a mixture of 5 mL of sulfuric acid R and 95 mL of ethanol System suitability Reference solution (d):
(96 per cent) R; heat at 110 °C for 20 min or until the spots — resolution', minimvim 4.5 between the peaks due to
appear. impurity C and cladribine.
System suitability: reference solution (b): Limits'.
— the chromatogram shows 2 clearly separated spots. — correction factors:for the calculation of content, multiply
Limit:. the peak areas of the following impurities by the
— impurity E: any spot due to impurity E is not more corresponding correction factor impurity B = 1.7;
intense than the spot in the chromatogram obtained with impurity C = 0.8;
reference solution (a) (0.3 per cent). — impurities A, C: for each impurity, not more than 3 times
the area of the principal peak in the chromatogram
Related substances obtained with reference solution (c) (0.3 per cent);
Liquid chromatography (2.2.29). — impurities B, D: for each impurity, not more than twice
Solvent mixture acetonitrile R, water R (10:90 VIV). the area of the principal peak in the chromatogram
Test solution (a) Dissolve 25.0 mg of the substance to be obtained with reference solution (c) (0.2 per cent);
examined in the solvent mixture and dilute to 5.0 mT, with — unspecified impurities: for each impurity, not more than the
the solvent mixture. area of the principal peak in the chromatogram obtained
Test solution (b) Dissolve 20.0 mg of the substance to be with reference solution (c) (0.10 per cent);
examined in the solvent mixture and dilute to 100.0 mT. with — total: not more than 10 times the area of the principal
the solvent mixture. peak in the chromatogram obtained with reference
solution (c) ( 1.0 per cent);
Reference solution (a) Dissolve 20.0 mg of cladribine CRS in
— disregard limit, the area of the principal peak in the
the solvent mixture and dilute to 100.0 mT. with the solvent
chromatogram obtained with reference solution (e)
mixture.
(0.05 per cent).
Reference solution (b) Dilute 1.0 mL of test solution (a) to
100.0 mL with the solvent mixture. W ater (2.5.32)
Maximum 0.5 per cent, determined on 0.100 g.
2016 Clarithromycin 1-577
DEFINITION
(3R,45,55,6R,7R,9R,nR, 12R, 135,14J?)-4-[(2,6-Dideoxy-3-
C-methyl-3-O-methyi-a-L-nJo-hexopyranosyl) oxy] -14-ethyl-
A. R = NH2: 9-(2-deoxy-ß-D-ö7yrÄn>-pentofuranosyl)-9H- 12,13-dihydroxy-7-methoxy-3,5,7,9,11,13-hexamethyl-6-
purin-2,6-diamine, [[3,4,6-trideoxy-3-(dimethylamino)-P-D-xyfo-
hexopyranosyl]oxy]oxacyclotetradecane-2 , l 0-dione (6 -0 -
B. R = OCH3: 9-(2-deoxy-ß-D-eo'£Äro-pentofuranosyl)-2-
methylerythromycin A).
methoxy-9H-purin-6-amine,
Semi-synthetic product derived from a fermentation product.
nh2
Content
n itA 96.0 per cent to 102.0 per cent (anhydrous substance).
Cl
a nI > CHARACTERS
Appearance
C. 2-chloro-7H-purin-6-amine (2-chloroadenine), White or almost white, crystalline powder.
Solubility
Practically insoluble in water, soluble in acetone and in
methylene chloride, slightly soluble in methanol.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison: clarithromycin CRS.
D. 2-chloro-9-(2-deoxy-a-i>go'£Äro-pentofiiranosyl)-9H- TESTS
purin-6 -amine, Solution S
HOv Dissolve 0.500 g in methylene chloride R and dilute to
50.0 mL with the same solvent.
OH
Appearance of solution
Solution S is clear or not more opalescent than reference
OH
suspension II (2.2.1) and not more intensely coloured than
E. 2 -deoxy-D-eryifcro-pentofuranose (2-deoxy-i>ribose), reference solution Y7 (2.2.2, Method II).
1-578 Clarithromycin 2016
R 3 -0 , ,0 —R
D. R l = R2 = R3 = H: 3"-N-demethyl-6-0-
methylerythromycin A,
ic + ir
E. Rl = R2 = CH3, R3 = H: 6,1 1-di-O-methylerythromycin Clebopride Malate ★ ' ★
★ ★
A,
(Ph. Eur. monograph 1303) *****
F. Rl = R3 = CH3, R2 = H: 6 , 12-di-O-methylerythromycin
A,
HOJ X »
H. Rl = CHO, R2 = R3 = H: 3 "-N-demethyl-3 '-iV-formyl-
6 -O-methylerythromycin A, a
■ H )
h o 2c
h 2n och3 and enantiomer
A. (li?5,2i?)-2-[2-[(li?)-l -(4-chlorophenyl)-1-
Time Mobile phase A Mobile phase B phenylethoxy] ethyl] -1 -methylpyrrolidine 1-oxide,
(min) (per cent V7V) (per cent V7V)
0-3 45 55
3 -2 3 45 -»5 55 -» 95
PH3 j ^ ^ N - CH3
23 - 26 5 95
pH (2.2.3)
Clenbuterol Hydrochloride }***%
5.0 to 7.0 for solution S.
(Ph. Eur. monograph 1409) * Optical rotation (2.2.7)
- 0 .10° to + 0 .10°.
V HH Dissolve 0.30 g in water R and dilute to 10.0 mL with the
ci\ / V v ny ch3 same solvent. Filter if necessary.
I J H3C CH3 and enantlomer , HCI
Related substances
Liquid chromatography (2.2.29).
Cl
Test solution Disperse 100.0 mg of the substance to be
examined in the mobile phase and dilute to 50.0 mL with
C 12H 1 9 Cl3N20 313.7 21898-19-1
the mobile phase.
Action and use Reference solution (a) Dilute 0.1 mL of the test solution to
Beta2-adrenoceptor agonist; bronchodilator. 100.0 mL with water R.
Reference solution (b) Dissolve 5 mg of clenbuterol
PhEur----------------------------------------------------------------------------------------
impurity B CRS in 10 mL of the mobile phase, add 2.5 mL
DEFINITION of the test solution and dilute to 25.0 mL with the mobile
(li?5)-l -(4-Amino-3j5-dichlorophenyl)-2-[(1,1 - phase.
dimethylethyl)amino]ethanol hydrochloride. Column:
Content — size: I = 0.125 m, 0 = 4 mm,
99.0 per cent to 101.0 per cent (anhydrous substance). — stationary phase: end-capped octadecylstlyl silica gel for
chromatography R (5 |im),
CHARACTERS — temperature: 40 °C.
Appearance
Mobile phase Mix 200 volumes of acetonitri"le R, 200 volumes
White or almost white, crystalline powder.
of methanol R and 600 volumes of a solution prepared as
Solubility follows: dissolve 3.0 g of sodium decanesulfonate R and 5.0 g
Soluble in water and in ethanol (96 per cent), slightly soluble of potassium dihydrogen phosphate R in 900 mL of water R,
in acetone. adjust to pH 3.0 with dilute phosphoric acid R and dilute to
mp 1000 mL with water R.
About 173 °C, with decomposition. Flow rate 0.5 mL/min.
IDENTIFICATION Detection Spectrophotometer at 215 nm.
First identification A, C. Injection 5 pL.
Second identification B, C. Run time 1.5 times the retention time of clenbuterol.
A. Infrared absorption spectrophotometry (2.2.24). Retention time Clenbuterol = about 29 min.
Comparison clenbuterol hydrochloride CRS. System suitability: reference solution (b):
B. Thin-layer chromatography (2.2.27). — resolution: minimum 4.0 between the peaks due to
impurity B and clenbuterol.
Test solution Dissolve 10 mg of the substance to be examined
in 10 mL of methanol R Limits:
— impurities A, B, C, D, E, F: for each impurity, not more
Reference solution Dissolve 10 mg of clenbuterol
than the area of the principal peak in the chromatogram
hydrochloride CRS in 10 mL of methanol R
obtained with reference solution (a) (0.1 per cent),
Plate T L C silica gel F 254 plate R — any other impurity, for each impurity, not more than the
Mobile phase ammonia R, anhydrous ethanol R, toluene R area of the principal peak in the chromatogram obtained
(0.15:10:15 VIVIV). with reference solution (a) (0 . 1 per cent),
Application 10 jiL. — total: not more than twice the area of the principal peak in
Development Over a path of 10 cm. the chromatogram obtained with reference solution (a)
(0.2 per cent),
Drying In air.
— disregard limit: 0.5 times the area of the principal peak in
Detection Spray with a 10 g/L solution of sodium nitrite R in the chromatogram obtained with reference solution (a)
1 M hydrochloric add and dip after 10 min in a 4 g/L solution (0.05 per cent).
of naphthylethylenediamme (¡¡hydrochloride R in methanol R
W ater (2.5.12)
Allow to dry in air.
Maximum 1.0 per cent, determined on 0.500 g.
Results The principal spot in the chromatogram obtained with
the test solution is similar in position, colour and size to the Sulfated ash (2.4.14)
principal spot in die chromatogram obtained with the Maximum 0.1 per cent, determined on 1.0 g.
reference solution. ASSAY
C. It gives reaction (a) of chlorides (2.3.1). Dissolve 0.250 g in 50 mL of ethanol (96 per cent) R and add
5.0 mL of 0.01 M hydrochloric add. Titrate with 0.1 M
TESTS
sodium hydroxide, determining the end-point
Solution S potentiometrically (2.2.20). Read the volume added between
Dissolve 0.5 g in 10 mL of carbon dioxide-free water R. the 2 points of inflexion.
Appearance of solution 1 mL of 0.1 M sodium hydroxide is equivalent to 31.37 mg of
Solution S is not more opalescent than reference Ci2H i 9Cl3N 20 .
suspension II (2.2.1) and not more intensely coloured than
reference solution Y6 (2.2.2, Method IT).
1-584 Clindamycin Hydrochloride 2016
IMPURITIES IDENTIFICATION
Specified impurities: A , B, C, D, E, F. First identification: A , D.
Second identification B , C, D
R2
A. Infrared absorption spectrophotometry (2.2.24).
R1 Comparison clindamycin hydrochloride CRS.
H2N
1 / B. Thin-layer chromatography (2.2.27).
R2 Test solution Dissolve 10 mg of the substance to be examined
in methanol R and dilute to 10 mL with the same solvent.
A. R l = H, R2 = Cl: 4-amino-3,5-dichlorobenzaldehyde, Reference solution (a) Dissolve 10 mg of cBndamycm
B. R l = CH 2-NH-C(CH 3)33 R2 = Cl: l-(4-amino-3,5- hydrochloride CRS in methanol R and dilute to 10 mL with the
dichlorophenyl)-2- [( 1,1 -dimethylethyl)amino] ethanone, same solvent.
C. R l = CH3, R2 = Cl: l-(4-amino-3,5- Reference solution (b) Dissolve 10 mg of clindamycin
dichlorophenyl) ethanone, hydrochloride CRS and 10 mg of lincomycin hydrochloride CRS
D. R l = CH3, R2 = H: 1-(4-aminophenyl)ethanone, in methanol R and dilute to 10 mL with the same solvent.
E. R l = CH 2Br, R2 = Cl: 1-(4-amino-3,5-dichlorophenyl)- Plate TLC sQica gel G plate R.
2 -bromoethanone, Mobile phase Mix 19 volumes of 2-propanol R, 38 volumes of
a 150 g/L solution of ammonium acetate R adjusted to pH 9.6
H OH with ammonia R, and 43 volumes of ethyl acetate R.
Application 5 jiL.
Development Over a path of 15 cm using the upper layer of
the mobile phase.
and enantiomer
Drying Id. air.
TESTS
pH (2.2.3)
CigH^C^Î^OgS 461.5 3.0 to 5.0.
21462-39-5
Dissolve 1.0 g in carbon dioxide-free water R and dilute to
Action and use 10 mL with the same solvent.
Lincosamide antibacterial. Specific optical rotation (2.2.7)
Preparation + 135 to + 150 (anhydrous substance).
Clindamycin Capsules Dissolve 1.000 g in water R and dilute to 25.0 mL with the
same solvent.
PhEur______________________
Related substances
DEFINITION Liquid chromatography (2.2.29).
Methyl 7-chloro-6, 7,8 -trideoxy-6 - [[[(2S,4R)-1 -methyl-4-
Test solution Dissolve 50.0 mg of the substance to be
propylpyrrolidin-2-yl] carbonyl] amino] -1 -xhio-L-threo-a-D-
examined in the mobile phase and dilute to 50.0 mL with
^a/aczo-octopyranoside hydrochloride. It contains a variable
the mobile phase.
quantity of water.
Reference solution (a) Dissolve 50.0 mg of clindamycin
Semi-synthetic product derived from a fermentation product
hydrochloride CRS in the mobile phase and dilute to 50.0 mL
Content with the mobile phase.
91.0 per cent to 102.0 per cent (anhydrous substance). Reference solution (b) Dilute 2.0 mL of the test solution to
CHARACTERS 100.0 mL with the mobile phase.
Appearance Column:
White or almost white, crystalline powder. — sizer. I = 0.25 m, 0 = 4.6 mm,
Solubility — stationary phase', octadecylsilyl silica gel for chromatography R
Very soluble in water, slightly soluble in ethanol (5 pm).
(96 per cent).
2016 Clindamycin Phosphate 1-585
18 -3 9 50 50
2016 Clindamycin Phosphate 1-587
B. methyl 7-chloro-6,7,8-trideoxy-6-[[[(2.S,4R)-4-ethyl-l-
methylpyrrolidin-2-yl] carbonyl] amino] -2-O-phosphono-1- G. methyl 6,8-dideoxy-2,4-0-(hydroxyphosphoryI)-6-
thio-L-iÄraHX-D-^oZacio-octopyranoside (clindamycin B [[[(25,4R)-1-methyl-4-propylpyrrolidin-2-yl] carbonyl] amino] -
2-phosphate), l-thio-i>^J3«Än>-a-D-^a/aca>-octopyranoside (2,4-phosphatidyl
lincomycin),
h a
H-
C. methyl 7-chloro-6,7,8-trideoxy-6-[[[(2.S,4R)-l-methyl-4-
propylpyrrolidin-2-yl] carbonyl] amino] -3-O-phosphono-1- H. methyl 7-chloro-6,7,8-trideoxy-6-[[ [(25,4R)- 1-methyl-4-
thio-L-i^aHX-D-^a/acio-octopyranoside (clindamycin propylpyrrolidin-2-yl]carbonyl]amino]-2,3-di-0-phosphono-l-
3-phosphate), thio-L-i^«HX-i>^a/acio-octopyranoside (clindamycin
2,3-bisphosphate),
H Cl
H-
/ /^
h3c ho o
D. methyl 7-chloro-6,7,8-trideoxy-6-[[[(2S,4R)-l-methyl-4-
propylpyrrolidin-2-yl] carbonyl] amino] -4-O-phosphono-1-
I. methyl 7-chloro-6,7,8-trideoxy-6- [[[(25,4R)-1-methyl-4-
thio-L-£Är«HX-i>^a/aczo-octopyranoside (clindamycin
propylpyrrolidin-2-yl] carbonyl] amino] -2,4-di-O-phosphono-1-
4-phosphate),
thio-L-rÄreo-a-D-^o/acro-octopyranoside (clindamycin
2,4-bisphosphate),
E. methyl 7-chloro-6,7,8-trideoxy-6-[[[(2.S,4R)-l-methyl-4-
propylpyrrolidin-2-yl] carbonyl] amino]-1-ûùo-L-threo-a-D-
gcdacto-octopyranoside (clindamycin), J. methyl 7-chloro-6j7,8-trideoxy-6-[[[(25)-l-methyl-4-
propylidenepyrrolidin-2-yl] carbonyl] amino] -2-O-phosphono-
1-xhio-L-threo-a-D-galacto-octopyxanosidc (propylidene analog
of clindamycin 2-phosphate),
1-588 Clioquinol 2016
H Cl Solubility
Practically insoluble in water, sparingly soluble in methylene
H* chloride, very slightly soluble or slightly soluble in ethanol
(96 per cent).
IDENTIFICATION
First identification: B.
Second identification A , C, D
A. Dissolve 40.0 mg in methanol R and dilute to 100.0 mL
K. 2,2 '-oxybis(hydroxyphosphoryl)bis [methyl 7-chloro-6,7,8- with the same solvent. Dilute 10.0 mL to 100.0 mL with
trideoxy-6- [ [[(2S,4R)~1-methyl-4-propyipyrroIidin-2- methanol R (solution A). Examined between 280 nm and
yl] carbonyl] amino] -1 -diio-L-iAreo-a-D-^a/aczo-octopyranoside] 350 nm (2.2.25), solution A shows an absorption maximum
(diclindamycin pyrophosphate), at 321 nm. Dilute 10.0 mL of solution A to 100.0 mL with
methanol R (solution B). Examined between 230 nm and
280 nm, solution B shows an absorption m a x im u m at
255 n m . The specific absorbance at this absorption
maximum is 1530 to 1660.
B. Infrared absorption spectrophotometry (2.2.24).
Preparation Discs of potassium bromide R.
Comparison clioquinol CRS.
C. When heated, violet fumes are produced.
D. Dissolve about 1 mg in 5 mL of ethanol (96 per cent) R.
Add 0.05 mL offerric chloride solution R l. A dark green
L. methyl 7-chloro-6 ,7,8-trideoxy-6- [[[(25,4R)-1-methyl-4- colour develops.
propylpyrrolidin-2 -yl] carbonyl] amino] - 2 -O-phosphono- 1-
TESTS
tMo-D-erythro-a-D-galacto-octopyranoside (7-epiclindamycin
2-phosphate). Acidity o r alkalinity
PhEur Shake 0.5 g with 10 mL of carbon dioxide-free water R and
filter. To the filtrate add 0.2 mL of phenolphthalein solution R.
The solution is colourless. Not more than 0.5 mL of 0.01 M
sodium hydroxide is required to change the colour of the
indicator to pink.
Clioquinol ★* * * ★ Related substances
★ ★
***** Liquid chromatography (2.2.29).
(Ph. Eitr. monograph 2111)
Test solution Dissolve 50.0 mg of the substance to be
examined in methanol R and dilute to 50.0 mL with the same
solvent, heating gently if necessary. Dilute 10.0 mL of the
solution to 25.0 mL with the mobile phase.
Reference solution (a) Dissolve 20.0 mg of 5-chloroquinolin-8-
ol R, 10.0 mg of 5,7-dichbroquinolin-S-ol R, 5 mg of the
substance to be examined and 10.0 mg of 5,7-diiodoquinolin-
8 -0 IR in methanol R, hearing gently if necessary and dilute to
C9H5CIINO 305.5 130-26-7 20.0 mL with the same solvent. Dilute 4.0 mL of the
solution to 50.0 mL with the mobile phase.
Action and use Reference solution (b) Dilute 1.0 mL of reference solution (a)
Antibacterial; antiprotozoal. to 10.0 mL with the mobile phase.
Preparations Reference solution (c) Dilute 1.0 mL of the test solution to
Betamethasone and Clioquinol Cream 100.0 mL with the mobile phase. Dilute 1.0 mL of this
Betamethasone and Clioquinol Ointment solution to 20.0 mL with the mobile phase.
Fhimetasone and Clioquinol Ear Drops Column:
Hydrocortisone and Clioquinol Cream — size. I = 0.15 m, 0 = 3.9 mm,
Hydrocortisone and Clioquinol Ointment — stationary phase: octylsOyl silica gelfor chromatography R
(5 pm).
PhEir. Mobile phase Dissolve 0.50 g of sodium edetate R in 350 mL of
DEFINITION water R, add 4.0 mL of hexylamine R and mix. Adjust to
5-Chloro-7-iodoquinolin-8-ol. pH 3.0 with phosphoric add R. Add 600 mL of methanol R
and dilute to 1000 mL with water R.
Content
Flow rate 1.3 mL/min.
98.0 per cent to 102.0 per cent (dried substance).
Detection Spectrophotometer at 254 nm.
CHARACTERS
Injection 20 pL.
Appearance
Almost white, light yellow, brownish-yellow or yellowish-grey Run time 4 times the retention time of clioquinol.
powder. Relative retention With reference to clioquinol (retention
time = about 10 min): impurity A = about 0.4;
iitipurity B = about 0.7; impurity C = about 1.3.
2016 Clobazam 1-589
Calculate the content of C 16H 13C1N 20 2 taking the specific Action and use
absorbance to be 1380. Glucocorticoid.
Preparations
IMPURITIES
Clobetasol Cutaneous Foam
Clobetasol Scalp Application
Clobetasol Shampoo
PhEur_________________________
DEFINITION
2 1-Chloro-9-fluoro-11 P-hydroxy-16|3-methyl-3,20-
dioxopregna-l,4-dien-l7-yi propanoate.
Content
A. R l = R3 = R4 = H, R2 = Cl: 7-chloro-5-phenyl-l,5- 97.0 per cent to 102.0 per cent (dried substance).
dihydro-3/f-1,5-benzodiazepme-2,4-dione, CHARACTERS
B. R l —CH3, R2 = R3 = R4 = H: 1-methyl-5-phenyl-1,5- Appearance
dihydro-3/i-1,5-benzodiazepine-2,4-dione, White or almost white, crystalline powder.
C. R l = R3 = CH3, R2 = Cl, R4 = H: (3J?S)-7-chloro-l,3- Solubility
dimethyl-5-phenyl-l,5-dihydro-3.£M,5-benzodiazepine-2,4- Practically insoluble in water, freely soluble in acetone,
dione, sparingly soluble in ethanol (96 per cent).
D. Rl = R3 = R4 = CH3, R2 = Cl: 7-chloro-1,3,3- IDENTIFICATION
trimethyi-5-phenyl- l,5-dihydro-3//-1,5-benzodiazepine-2,4- Infrared absorption spectrophotometry (2.2.24).
dione,
Comparison clobetasol propionate CRS.
TESTS
Specific optical rotation (2.2.7)
+ 112 to + 118 (dried substance).
Dissolve 0.500 g in acetone R and dilute to 50.0 mL with the
same solvent
Related substances
Liquid chromatography (2.2.29).
Test solution (a) Dissolve 20.0 mg of the substance to be
E. N- [4-chloro-2-(phenylamino)phenyl]-N-methylacetamide, examined in the mobile phase and dilute to 20.0 mL with
the mobile phase.
Test solution (b) Dissolve 20.0 mg of the substance to be
examined in the mobile phase and dilute to 100.0 mL with
the mobile phase.
2016 Clobetasol Propionate 1-591
Reference solution (a) Dissolve 20.0 mg of dobetasol — unspecified impurities:, for each impurity, not more than
propionate CRS in the mobile phase and dilute to 100.0 mL 0.2 times the area of the principal peak in the
with the mobile phase. chromatogram obtained with reference solution (d)
Reference solution (b) Dissolve the contents of a vial of (0.10 per cent);
clobetasol impurity J CRS in 2.0 mL of the mobile phase. — total: not more than 4 times the area of the principal peak
To 0.5 mL of this solution add 0.5 mL of test solution (b) in the chromatogram obtained with reference solution (d)
and dilute to 20.0 mL with the mobile phase. (2.0 per cent);
Reference solution (c) Dissolve the contents of a vial of
— disregard Umir. 0.1 times the area of the principal peak in
cbbetasolfor peak identification CRS (containing impurities
the chromatogram obtained with reference solution (d)
A, B, C, D, E, L and M) in 2 mL of the mobile phase. (0.05 per cent).
Reference solution (d) Dilute 1.0 mL of test solution (a) to Loss on drying (2.2.32)
50.0 mL with the mobile phase. Dilute 5.0 mL of this Maximum 0.5 per cent, determined on 1.000 g by drying in
•solution to 20.0 mL with the mobile phase. an oven at 105 °C for 3 h.
Column'. Sulfated ash (2.4.14)
— sizer. I = 0.15 m, 0 = 4.6 mm; Maximum 0.1 per cent, determined on 1.0 g.
— stationary phase: spherical octadecylsUyl silica gelfor ASSAY
chromatography R (5 pm); Liquid chromatography (2.2.29) as described in the test for
— temperature: 30 °C. related substances with the following modification.
Mobile phase Mix 10 volumes of methanol R, 42.5 volumes of Injection Test solution (b) and reference solution (a).
a 7.85 g/L solution of sodium dihydrogen phosphate
Calculate the percentage content of C 25H 32CIFO5 using the
monohydrate R adjusted to pH 5.5 with a 100 g/L solution of
chromatogram obtained with reference solution (a) and the
sodium hydroxide R and 47.5 volumes of acetonitrile R.
declared content of clobetasol propionate CRS.
Flow rate 1.0 mL/min.
STORAGE
Detection Spectrophotometer at 240 nm.
Protected from light.
Injection 10 pL of test solution (a) and reference
solutions (b)3 (c) and (d). IMPURITIES
Run time 3 times the retention time of clobetasol propionate. Specified impurities A,B, C, D, E, L, M
Identification of impurities Use the chromatogram supplied Other detectable impurities (the following substances would, if
with clobetasolfor peak identification CRS and the present at a sufficient level, be detected by one or other of
chromatogram obtained with reference solution (c) to identify the tests in the monograph. They are limited by the general
the peaks due to impurities A, B, C, D, E, L and M. acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
Relative retention With reference to clobetasol propionate
(2034). It is therefore not necessary to identify these
(retention time = about 10 min): impurity A = about 0.4;
impurities for demonstration of compliance. See also 5.10.
impurity B = about 0.6; impurity C = about 0.9;
Control of impurities in substancesfor pharmaceutical use): F, G,
impurity J = about 1.1; impurity D = about 1.2; H ,I ,J ,K .
impurity L = about 1.3; impurity M = about 1 .6 ;
impurity E = about 2.1.
System suitability:
— resolution: minimum 2.0 between the peaks due to
clobetasol propionate and impurity J in the chromatogram
obtained with reference solution (b);
— the chromatogram obtained with reference solution (c) is
similar to the chromatogram supplied with cbbetasolfor
peak identification CRS .
Limits: A. RI = CO-C2HS, R2 = OH: 9-fiuoro-1113,21-dihydroxy-
— correction factors: for the calculation of content, multiply 16fi-methyl-3,20-dioxopregna-l ,4-dien- 17-yl propanoate
the peak areas of the following impurities by the (betamethasone 17-propionate),
corresponding correction factor impurity B = 0.6; G. RI = H, R2 = Cl: 21-chloro-9-fluoro-11P, 17-dihydroxy-
impurity C = 1.5; 16P-methylpregna-l ,4-diene-3,20-dione (clobetasol),
— impurity E: not more than 1.4 times die area of the
principal peak in the chromatogram obtained with H. R1 = CO-C2H5, R2 = H: 9-fluoro-1 1^-hydroxy-16{i-
reference solution (d) (0.7 per cent); methyl-3,20-dioxopregna-1,4-dien-17-yl propanoate,
— impurity D: not more than the area of the principal peak I. R1 = CO-C2H5, R2 = 0-S0 2-CH3: 9-fluoro-lip-
in the chromatogram obtained with reference solution (d) hydroxy-16P-methyl-2 1- [(methylsulfonyl) oxy]-3,20-
(0.5 per cent); dioxopregna-1,4-dien-17-yl propanoate,
— impurities B, C: for each impurity, not more than K. RI = H, R2 = 0-C0-C 2H5: 9-fluoro-11P, 17-dihydroxy-
0.6 times the area of the principal peak in the 16P-methyl-3,20-dioxopregna-1,4-dien-21-yl propanoate
chromatogram obtained with reference solution (d) (betamethasone 21 -propionate),
(0.3 per cent);
— impurities A , L, M : for each impurity, not more than
0.4 times the area of the principal peak in the
chromatogram obtained with reference solution (d)
(0.2 per cent);
1-592 Clobetasone Butyrate 2016
Wl
/0
Cl
°s J o
F. 9-fluoro-l 1P-hydroxy-16P-methyl-3-oxopregna-l ,4,17 (20)- Solvent mixture anhydrous formic acid R, acetonitrUe R, water R
trien-21-oic add, (0.1:43:57 V/V/V).
Test solution Dissolve 65 mg of the substance to be examined
in 5.0 mL of acetomtrile R and dilute to 25.0 mL with die
solvent mixture.
Reference solution (a) Dissolve 13 mg of clobetasone butyrate for
system suitability CRS (containing impurity F) in 1 mL of
acetonitrile R and dilute to 5.0 mL with the solvent mixture.
Reference solution (b) Dilute 1.0 mL of the test solution to
100.0 mL with the solvent mixture. Dilute 1.0 mL of this
solution to 10.0 mL with the solvent mixture.
J. (ni^^'-chloro-S'-ethyW -fluoro-l 1P-hydroxy-160-
Column:
methylspiro [androsta-1,4-diene-17,2' (3 7f)-furan]-3,3 '-dione
— sizer. I = 0.15 m, 0 = 4.6 mm;
(17a-spiro compound),
— stationary phase, end-capped octadecylsifyl silica gelfor
L. unknown structure, chromatography R (3.5 pm);
M. unknown structure. — temperature: 40 °C.
_________ :__________________________ PhEur
2016 Clobetasone Butyrate 1-593
Mobile phase:
— mobile phase A: anhydrous formic acid R, water R
(0.1:99.9 VIV);
— mobile phase B: anhydrous formic add R, acetomtrUe R
(0.1:99.9 VIV)',
Me
c S 'OH
S
.OH
// \\
vmylthiazole edisilate BPCRS in methanol (solution A) to
50 volumes with the mobile phase. For solution (4) dilute
1 volume of a 0.020% w/v solution of 5-(2-chloroethyl)-4-
o o
methyl-3-[2-(4-methylthiazol-5-yl)ethyl]-tkiazolium
chloride BPCRS (quaternary dimer) in methanol (solution B)
(CeHsClNS^CiHftOeSa 513.5 1867-58-9 to 50 volumes with the mobile phase. For solution (5) dilute
1 volume of a 0.020% w/v solution of 4-methyl-5-
Action and use (2-hydroxyethyl)thiazole BPCRS in methanol (solution C) to
Hypnotic. 50 volumes with the mobile phase. For solution (6) add
Preparations 1 mL each of solutions A, B and C to 0.15 g of the
Clomethiazole Infusion substance being examined and dilute to 50 mL with the
Clomethiazole Oral Solution mobile phase.
The chromatographic procedure may be carried out using
DEFINITION (a) a stainless steel column (20 cm x 4 mm) packed with
Clomethiazole Edisilate is 5-(2-chloroethyl) -4-methylthiazole octadecylsUyl silica gelfar chromatography (10 pm) (Iichrosorb
ethanedisulfonate. It contains not less than 99.0% and not RP18 is suitable), (b) as the mobile phase with a flow rate of
more than 101.0% of (C6H8dN S) 2,C2H60 6S2, calculated 1 mL per minute, a mixture of 70 volumes of a solution
with reference to the dried substance. containing 0.13% w/v of sodium hexanesidfonate and 2.7% w/v
CHARACTERISTICS of tetramethylammonium hydrogen sulfate, adjusted to pH 2.0
A white, crystalline powder. with 5m sodium hydroxide, and 30 volumes of methanol and
(c) a detection wavelength of 257 nm.
Freely soluble in water, soluble in ethanol (96%); practically
insoluble in ether.
1-598 Clomifene Citrate 2016
The test is not valid unless in the chromatogram obtained Preparation Discs of potassium bromide R.
with solution (6) baseline separation is achieved between the Comparison domifene citrate CRS.
peaks due to the three specified impurities and also between B. Dissolve about 5 mg in 5 mL of a mixture of 1 volume of
the principal peak and the two adjacent specified acetic anhydride R and 5 volumes of pyridine R, then heat in a
impurity peaks. water-bath. A deep red colour is produced.
Calculate the content of each of the three specified impurities
TESTS
in the substance being examined with reference to
Prepare the solutions protectedfrom light in brown-glass vessels
clomethia2ole base (1 mg of clomethiazole edisilate is
Ensure minimum exposure of the solutions to daylight until they
equivalent to 0.629 mg of base) expressing the content of
are requiredfor chromatography.
4-methyl-5-vinylthiazole as the base (1 mg of 4-methyl-5-
vinylthiazole edisilate is equivalent to 0.568 mg of base). Related substances
The total content of the three specified impurities is not Liquid chromatography (2.2.29).
greater than 0.5%. In the chromatogram obtained with Test solution Dissolve 12.5 mg of the substance to be
solution (1) the area of any other secondary peak is not greater examined in the mobile phase and dilute to 10.0 mL with
than the area of the peak in the chromatogram obtained with the mobile phase.
solution (2). Reference solution (a) Dissolve 12.5 mg of domifene citrate for
Loss on drying performance test CRS in the mobile phase and dilute to
When dried at 50° at a pressure not exceeding 0.7 kPa for 10.0 mL with the mobile phase.
6 hours, loses not more than 0.5% of its weight. Use 1 g. Reference solution (b) Dilute 1.0 mL of the test solution to
Sulfated ash 50.0 mL with the mobile phase.
Not more than 0.3%, Appendix IX A. Use 1 g. Column:
ASSAY — size. I = 0.25 m, 0 = 4.6 mm;
Dissolve 0.4 g in 50 mL of water and titrate with 0.1m sodium — stationary phase, butylsüyl silica gd for chromatography R
IMPURITIES a
Specified impurities A, 8, C, D, E, F, G, H
, Ar = = . Ha
ch3
17321-77-6
Ar*
Ar and (ZHsomer Action and use
Ar Monoamine reuptake inhibitor; tricyclic antidepressant.
Preparation
A. 2- [4-( 1,2-diphenylethenyl)phenoxy] -NJsf- Clomipramine Capsules
diethylethanamine, Prolonged-release Clomipramine Tablets
1-600 2016
ci
— impurity C} D". for each impurity, not more than the area
of the corresponding peak in the chromatogram obtained
with reference solution (a) (0.2 per cent), . ch3
— impurity F: not more than the area of the corresponding N
I
peak in the chromatogram obtained with reference ch3
solution (a) (0.1 per cent),
— arty other impurity, not more than 0.1 times the area of the
principal peak in the chromatogram obtained with
reference solution (b) (0.1 per cent), C. 3-(3-chloro-5//-dibenzo [bj\azepin-5-yl)-iV^V-
— total of other impurities: not more than 0.2 times the area dimethylpropan- 1-amine,
of the principal peak in the chromatogram obtained with
reference solution (b) (0.2 per cent), R1
— total: not more than the area of the principal peak in the
• chromatogram obtained with reference solution (b)
( 1.0 per cent),
— disregard limit. 0.01 times the area of the principal peak in
the chromatogram obtained with reference solution (b)
(0.01 per cent). R3
Heavy metals (2.4.8)
Maximum 20 ppm. D. RI = R3 = Cl, R2 = CH2-CH 2-CH 2-N(CH3)2: 3-(3,7-
2.0 g complies with test C. Prepare the reference solution dichloro-10,11 -dihydro-5//-dibenzo [bj\ azepin-5-yl)-N,N-
using 4 mL of lead standard solution (10 ppm Pb) R. dimethylpropan-1-amine,
Loss on drying (2.2.32) E. RI = R2 = R3 = H: 10,ll-dihydro-5tf-
Maximum 0.5 per cent, determined on 1.000 g by drying in dibenzo [bj\ azepine (iminodibenzyl),
an oven at 105 °C. F. RI = Cl, R2 = R3 = H: 3-chloro-10,l l-dihydro-5ii-
Sulfated ash (2.4.14) dibenzo [bj\azepine,
Maximum 0.1 per cent, determined on 1.0 g. G. RI = Cl, R2 = CH2-CH=CH 2, R3 = H: 3-chloro-5-
ASSAY (prop-2-enyl)-l 0,11 -dihydro-5ii-dibenzo [bj\azepine.
Dissolve 0.250 g in 50 mL of alcohol R and add 5.0 mL of ______________________________________________________ ___ PhEur
0.01 M hydrochloric acid. Carry out a potentiometric titration
(2.2.20), using 0.1 M sodium hydroxide. Read the volume
added between the 2 points of inflexion.
1 mL of 0.1 M sodium hydroxide is equivalent to 35.13 mg of ★ ★
C19H 24CI2N 2.
Clonazepam ★ ★
STORAGE (Ph. Eur. monograph 0890) *****
In an airtight container, protected from light.
IMPURITIES
ci
.CH3
N N
1 1
ch3 ch3
DEFINITION
5-(2-Chlorophenyl)-7-nitro-l,3-dihydro-2/f-l,4-
benzodiazepin-2 -one.
B. 3-(10,l 1-dihydro-5/i-dibenzo [bf\20£pm-5-yl)-N,N-
dimethylpropan- 1-amine (imipramine), Content
99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
Appearance
Slightly yellowish, crystalline powder.
1-602 Clonidine Hydrochloride 2016
Limits-.
— impurity A : not more than the area of the principal peak C9HJ0Q 3N 3 266.6 4205-91-8
in the chromatogram obtained with reference solution (a)
Action and use
(0.1 per cent),
Alpha2-adrenoceptor agonist; treatment of hypertension.
— impurity B: not more than the area of the principal peak
in the chromatogram obtained with reference solution (c) Preparations
(0.1 per cent) Clonidine Injection
— any other impurity, for each impurity, not more than the Clonidine Tablets
area of the principal peak in the chromatogram obtained
with reference solution (a) (0.1 per cent), PhEur__ ____________________________________________________ __
— total: not more than twice the area of the principal peak in DEFINITION
the chromatogram obtained with reference solution (a) 2,6-Dichloro-N-(imidazolidin-2-ylidene)aniline hydrochloride.
(0.2 per cent),
Content
— disregard lima: 0.5 times the area of the principal peak in
98.5 per cent to 101.0 per cent (dried substance).
the chromatogram obtained with reference solution (a)
(0.05 per cent). CHARACTERS
Appearance
White or almost white, crystalline powder.
2016 Clonidine Hydrochloride 1-603
çOo I H o>« c h 3
examined in
solvent.
methanol R and dilute to 10.0 mL with the same
MobUe phase anhydrous ethanol R, heptane R (15:85 VIV). Relative retention With reference to clopidogrel (retention
Flow rate 0.8 mL/min. time = about 25 min): impurity A = about 0.4;
impurity B = about 1.1.
Detection Spectrophotometer at 220 nm.
System suitability: reference solution (b):
Injection 10 pL.
— peak-to-valley ratio: minimum 10, where Hp — height
Run time 1.25 times the retention time of clopidogrel. above the baseline of the peak due to impurity B and
Identification of impurities Use die chromatogram supplied H v = height above the baseline of the lowest point of the
with clopidogrelfor system suitability CRS and the curve separating this peak from the peak due to
chromatogram obtained with the reference solution to clopidogrel.
identify the peaks due to impurities B and C. Limits:
Relative retention With reference to clopidogrel (retention — impurity B: not more than 3 times the area of the
time = about 18 min): impurity C = about 0.6; principal peak in the chromatogram obtained with
impurity B = about 0.7. reference solution (c) (0.3 per cent);
System suitability: reference solution: — impurity A: not more than twice the area of the principal
— resolution: minimum 2.0 between the peaks due to peak in the chromatogram obtained with reference
impurities C and B; solution (c) (0.2 per cent);
— signal-to-noise ratio: minimum 20 for the peak due to — unspecified impurities: for each impurity, not more than the
impurity C. area of the principal peak in the chromatogram obtained
Limit: with reference solution (c) (0.10 per cent);
— impurity C: maximum 0.5 per cent. — total: not more than 5 times the area of the principal peak
in the chromatogram obtained with reference solution (c)
Related substances
(0.5 per cent);
Liquid chromatography (2.2.29).
— disregard limit. 0.5 times the area of the principal peak in
Solvent mixture Mobile phase A, acetonitrile R1 (40:60 VIV). the chromatogram obtained with reference solution (c)
Test solution Dissolve 65 mg of the substance to be examined (0.05 per cent).
in the solvent mixture and dilute to 10.0 mL with the solvent Heavy metals (2.4.8)
mixture. Maximum 20 ppm.
Reference solution (a) Dissolve 5 mg of clopidogrel
1.0 g complies with test C. Prepare the reference solution
impurity A CR S in the solvent mixture and dilute to 25.0 mL
using 2 mL of lead standard solution (10 ppm Pb) R.
with the solvent mixture.
W ater (2.5.12)
Reference solution (b) Dissolve 32 mg of clopidogrelfor system
Maximum 0.5 per cent, determined on 1.00 g.
suitability CRS (containing impurities B and C) in the solvent
mixture, add 0.5 mL of reference solution (a) and dilute to Replace the solvent after each titration.
5.0 mL with the solvent mixture. Sulfated ash (2.4.14)
Reference solution (c) Dilute 1.0 mL of the test solution to Maximum 0.1 per cent, determined on 1.0 g.
100.0 mL with the solvent mixture. Dilute 1.0 mL of this ASSAY
solution to 10.0 mL with the solvent mixture. Dissolve 0.160 g in a mixture of 10 mL of acetone R, 10 mL
Column: of methanol R and 30 mL of water R. Titrate with 0.1 M
— size: I = 0.15 m, 0 = 3.9 mm; sodium hydroxide, determining the end-point
— stationary phase: end-capped octadecylsUyl silica gelfor potentiometrically (2.2.20). A precipitate may be formed
chromatography R (5 pm); during the titration.
— temperature: 30 °C. 1 mL of 0.1 M sodium hydroxide is equivalent to 20.99 mg of
Mobile phase: c 16h 18o n o 6s 2.
— mobile phase A: mix 5 volumes of methanol R2 and
95 volumes of a 0.96 g/L solution of sodium STORAGE
pentanesulfonate monohydrate R adjusted to pH 2.5 with
Protected from light.
phosphoric acid R; IMPURITIES
— mobile phase B: methanol R2, acetonitrile R1 (5:95 VIV); Specified impurities A,B, C
Other detectable impurities(die following substances would, if
Time Mobile phaae A Mobile phase B present at a sufficient level, be detected by one or other of
(min) (per cent V/V) (per cent V/V) the tests in the monograpB. They are limited by the general
0 -3 89.5 10.5
acceptance criterion for other/unspecified impurities and/or
3 -4 « 89.5 •» 315 10.5 •» 68.5 by the general monograph Substances for pharmaceutical use
4 8 -6 8 31.5 68.5
(2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): D.
Flow rate 1.0 mL/min.
Detection Spectrophotometer at 220 nm.
Injection 10 pL of the test solution and reference solutions (b)
and (c).
Identification of impurities Use the chromatogram supplied
with clopidogrelfor system suitability CRS and the
chromatogram obtained with reference solution (b) to
identify the peaks due to impurities A and B. A. (2¿)-(2-chlorophenyl) [6,7-dihydrothieno [3,2-c]pyridin-
5 (4/f)-ylj acetic acid,
2016 Clotrimazole 1-607
IDENTIFICATION
First identification B
Second identification A , C
A. Melting point (2.2.14): 141 °C to 145 °C.
B. Infrared absorption spectrophotometry (2.2.24).
B. methyl (25)-(2-chlorophenyl)[4,7-dihydrothieno[2,3- Comparison clotrimazole CRS.
c]pyridin-6 (5H)-yl] acetate. C. Thin-layer chromatography (2.2.27).
Test solution Dissolve 50 mg of the substance to be examined
in ethanol (96 per cent) R and dilute to 5 mL with the same
solvent.
och3 Reference solution Dissolve 50 mg of clotrimazole CRS in.
ethanol (96 per cent) R and dilute to 5 mL with the same
solvent.
Plate TLC silica gel F 254 plate R
C. methyl (2i?)-(2-chlorophenyl) [6,7-dihydrothieno[3,2-
Mobile phase concentrated ammonia R l, propanol R, toluene R
c]pyridin-5 (4H)~yI] acetate,
(0.5:10:90 VIV/V).
Application 10 (iL.
Development Over 2/3 of the plate.
Drying In air.
Detection Examine in ultraviolet light at 254 run.
Results The principal spot in the chromatogram obtained with
the test solution is similar in position and size to the principal
spot in the chromatogram obtained with the reference
D. methyl (2i?)-(2-chlorophenyl) [(23}-(2-chlorophenyl) [6,7- solution.
dihydrothieno [3,2-c]pyridin-5 (4H)-yl] acetyloxy] acetate.
TESTS
_______________________________________________ PhEur
Related substances
Liquid chromatography (2.2.29).
Test solution Dissolve 50.0 mg of the substance to be
★★*★ ★ examined in acetomtrUe R l and dilute to 50.0 mL with the
Clotrimazole ★ ★ same solvent.
(Ph. Eicr. monograph 0757) ***** Reference solution (a) Dilute 1.0 mL of the test solution to
100.0 mL with acetonitrUe R l. Dilute 1.0 mL of this solution
to 10.0 mL with acetonitrUe R l.
Reference solution (b) Dissolve the contents of a vial of
clotrimazole for peak identification CRS (containing impurities
A, B and F) in 1.0 mL of acetonitrUe R l.
Reference solution (c) Dissolve 5.0 mg of imidazole CRS
(impurity D) and 5.0 mg of clotrimazole impurity E CRS in
acetonitrUe R l and dilute to 100.0 mL with the same solvent.
C22Hi7CIN2 23593-75-1 Dilute 1.0 mL of this solution to 25.0 mL with
acetonitrUe R l.
Action and use
Column'.
Annfungal.
— sizer. I = 0.15 m, 0 = 4.6 mm;
Preparations — stationary phase', spherical end-capped octylsUyl sUica gel for
Clotrimazole and Hydrocortisone Acetate Cream chromatography R (5 |im);
Clotrimazole Cream — temperature: 40 °C.
Clotrimazole Eye Drops Mobile phase:
Clotrimazole Pessaries — mobile phase A: dissolve 1.0 g of potassium dihydrogen
phosphate R and 0.5 g of tetrabutylammomum hydrogen
Clotrimazole Vaginal Tablets
sulfate R l in water R and dilute to 1000 mL with the
PhEur. same solvent;
— mobile phase B: acetonitrUe Rl',
DEFINITION
1- [(2-Chlorophenyl) diphenylmethyl] - lH-imidazole.
Time Mobile phase A Mobile phase B
Content (min) (per cent V/V) (per cent V/V)
98.5 per cent to 100.5 per cent (dried substance). 0 -3 75 25
CHARACTERS 3 -2 5 75-»20 2 5 -»80
A ppearance 2 5 -3 0 20 80
White or pale yellow, crystalline powder.
Solubility
Flow rate 1.0 mL/min.
Practically insoluble in water, soluble in ethanol (96 per cent)
and in methylene chloride. Detection Spectrophotometer at 210 nm.
1-608 Cloxacillin Sodium 2016
Injection 10 |iL.
Relative retention With reference to clotrimazole (retention
time = about 12 min): impurity D = about 0.1;
impurity F = about 0.9; impurity B = about 1.1;
impurity E = about 1.5; impurity A = about 1.8.
System suitability: reference solution (b):
— resolution', minimum 1.5 between the peaks due to
impurity F and clotrimazole; B. R = Cl: l-[(4-chlorophenyl)diphenylmethyl]-lif-
— the chromatogram obtained is similar to the imidazole,
chromatogram supplied with clotrimazolefor peak F. R = H: l-(triphenylmethyl)-lif-imidazole
identification CRS. (deschloroclotrimazole),
Limits:
— impurities A , B: for each impurity, not more than twice
the area of the principal peak in the chromatogram
obtained with reference solution (a) (0.2 per cent);
— impurities D, E: for each impurity, not more than the area
of the corresponding peak in the chromatogram obtained D. imidazole.
with reference solution (c) (0.2 per cent); PhEur
— impurity F: not more than the area of the principal peak
in the chromatogram obtained with reference solution (a)
(0.1 per cent);
— unspecified impurities: for each impurity, not more than the
area of the principal peak in the chromatogram obtained Cloxacillin Sodium ★★* + +
★ +
with reference solution (a) (0.10 per cent);
(Ph. Eur. monograph 0661) *****
— total: not more than 5 times the area of the principal peak
in the chromatogram obtained with reference solution (a)
(0.5 per cent);
— disregard limit: 0.5 times die area of the principal peak in
the chromatogram obtained with reference solution (a) h 2o
(0.05 per cent).
Loss on drying (2.2.32)
Maximum 0.5 per cent, determined on 1.000 g by drying in
an oven at 105 °C.
Sulfated ash (2.4.14) CujHnClNaNaOsSjHjO 475.9 7081-44-9
Maximum 0.1 per cent, determined on 1.0 g.
Action and use
ASSAY Penicillin antibacterial.
Dissolve 0.300 g in 80 mL of anhydrous acetic acid R. Using
0.3 mL of naphtholbenzem solution R as indicator, titrate with PhEur_____________________________
0.1 M perchloric acid until the colour changes from brownish-
DEFINITION
yellow to green.
Sodium (2S,5R,6i?)-6- [[[3-(2-chlorophenyl)-5-
1 mL of 0.1 M perchloric acid is equivalent to 34.48 mg methylisoxazol-4-yl] carbonyl] amino]-3,3-dimethyl-7-oxo-4-
of C 22H 17CIN2. thia-l-azabicyclo[3.2.0]heptane-2-carboxylate monohydrate.
STORAGE Semi-synthetic product derived from a fermentation product.
Protected from light.
Content
IMPURITIES 95.0 per cent to 102.0 per cent (anhydrous substance).
Specified impurities A, B, D, E, F
CHARACTERS
Other detectable impurities (die following substances would, if Appearance
present at a sufficient level, be detected by one or other of White or almost white, hygroscopic, crystalline powder.
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or Solubility
by the general monograph Substancesfor pharmaceutical use Freely soluble in water and in methanol, soluble in ethanol
(2034). It is therefore not necessary to identify these
(96 per cent).
impurities for demonstration of compliance. See also 5.10. IDENTIFICATION
Control of impurities in substancesfor pharmaceutical use): C. First identification A , D
Second identification B, C, D
A. Infrared absorption spectrophotometry (2.2.24).
Preparation Discs.
Comparison cloxacillin sodium CRS.
A. R = OH, R' = C$H5: (2-dilorophenyi)diphenylmethanol, B. Thin-layer chromatography (2.2.27).
C. R = Cl, R ' = C$H5: l-chloro-2- Test solution Dissolve 25 mg of the substance to be examined
(chlorodiphenylmethyl) benzene, in 5 mL of water R.
E. R + R' = O: (2-chlorophenyl)phenylmethanone Reference solution (a) Dissolve 25 mg of cloxacillin sodium CRS
(2-chlorobenzophenone), in 5 mL of water R.
2016 Cloxacillin Sodium 1-609
Solubility
Practically insoluble in water, freely soluble in methylene
chloride, soluble in ethanol (96 per cent). It dissolves in
H H dilute acetic add.
IDENTIFICATION
C. (2S,5Ä,6i^-6-amino-3,3-dimethyI-7-oxo-4-thia-1- A. Mdting point (2.2.14): 182 °C to 186 °C.
azabicydo[3.2.0]heptane-2-carboxylic add
(6-aminopenirillanic add), B. Infrared absorption spectrophotometry (2.2.24).
Comparison clozapine CRS.
TESTS
Related substances
Liquid chromatography (2.2.29).
Solvent mixture water R, methanol R2 (20:80 V/V).
Solution A Dissolve 2.04 g of potassium dihydrogen phosphate R
in 1000 mL of water R and adjust to pH 2.4 ± 0.05 with
dilute phosphoric acid R.
D. 3-(2-chIorophenyi)-5-methylisoxazole-4-carboxylic add, Test solution Dissolve 75 mg of the substance to be examined
in 80 mL of methanol R2 and dilute to 100 mL with water R
o • »co2h Reference solution (a) Dilute 1.0 mL of the test solution to
V T ' V 0”3 10.0 mL with the solvent mixture. Dilute 1.0 mL of this
Cl— \ \ ° ^ n - - —L / X H a solution to 100.0 mL with the solvent mixture.
/ — 0 J - 'H H H Reference solution (b) Dissolve the contents of a vial of
N =( ^^N ^\.C H 3 clozapine for peak identification CRS (containing
O y X , N -i—Lg/^CHs impurities A, B, C and D) in 1.0 mL of the solvent mixture.
T H H H Column:
CH3 o
— size. I = 0.125 m, 0 = 4.6 mm;
— stationary phase, end-capped octadecylsUyl silica gelfor
E. (2S,5R,6R)-6- [[[(25,5R,6.R)-6-[ [[3-(2-chlorophenyl)-5- chromatography R (5 jjtm).
methyUsoxazol-4-yl]carbonyl]amino]-3,3-dimethyl-7-oxo-4-
Mobile phase.
thia-1-azabicydo [3.2.0]hept-2-yl] carbonyl] amino] -3,3-
— mobile phase A: acetonitrile for chromatography R,
dimethyl-7-oxo-4-thia-1 -azabicyclo [3.2.0] heptane-2-
methanol R2, solution A (1:1:8 V/V/V);
carboxylic add (6-APA cloxadllin amide).
— mobile phase B: acetonitrilefor chromatography R,
__________________________________________________________ PhEur methanol R2, solution A (4:4:2 V/V/V)',
nh2
— Z>. for each impurity, not more than twice
impurities B,
the area of the principal peak in the chromatogram
obtained with reference solution (a) (0.2 per cent); nh o
— impurity C: not more than 3 times the area of the
principal peak in the chromatogram obtained with
reference solution (a) (0.3 per cent); 6 ^ 0 . CH,
— unspecified impurities: for each impurity, not more than the
area of the principal peak in the chromatogram obtained D . 1- [2- [(2-amino-4-chlorophenyl) amino]benzoyI] -4-
with reference solution (a) (0.10 per cent); methylpiperazine.
— total: not more than 6 times the area of the principal peak PhEur
in the chromatogram obtained with reference solution (a)
(0.6 per cent);
— disregard limir. 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (a)
(0.05 per cent). Cocaine
Heavy metals (2.4.8)
Maximum 20 ppm.
1.0 g complies with test C. Prepare the reference solution
using 2 m L of lead standard solution (10 ppm Pb) R.
Loss on drying (2.2.32)
Maximum 0.5 per cent, determined on 1.000 g by drying in
an oven at 105 °C.
Sulfated ash (2.4.14)
Maximum 0.1 per cent, determined on 1.0 g.
ASSAY
Dissolve 0.100 g in 50 mL of anhydrous acetic acid R. Titrate
with 0.1 M perchloric arid, determining the end-point
potentiometrically (2.2.20). C17H2iN04 303.4 50-36-2
1 mL of 0.1 M perchloric add is equivalent to 16.34 mg
of CigHjgCIN^ Action and use
Local anaesthetic.
IMPURITIES
Specified impurities A, B, C, D. DEFINITION
Cocaine is methyl (1R,2R,3S,5S)-3-(benzoyloxy)-8-methyl-8-
azabicyclo[3.2.1]octane-2-carboxylate and may be obtained
from the leaves of Erythroxyhm coca Lam. and other species
of Erythroxyhim or by synthesis. It contains not less than
98.0% and not more than 101.0% of C 17H 2iN 0 4 , calculated
with reference to the dried substance.
CHARACTERISTICS
A. 8-chloro-5,l0-dihydro-1lH-dibenzo[V] [l,4]diazepin-l 1- Colourless crystals or a white, crystalline powder. Slightly
one, volatile.
Practically insoluble in water, freely soluble in ethanol (96%)
and in ether, soluble in arachis oil; slightly soluble in liquid
paraffin.
IDENTIFICATION
The infrared absorption spectrum, Appendix II A, is concordant
with the reference spectrum of cocaine (RS 071).
TESTS
Melting point
96° to 98°, Appendix V A.
Specific optical rotation
B. 11,11 '-(piperazine-1,4-diyl)bis(8-chloro-5//- In a 2.4 % w/v solution in 0.1m hydrochloric add, —79 to
dibenzo[t,e] [l,4]diazepine), -8 1 , calculated with reference to the dried substance,
Appendix V F.
a NH Related substances
Carry out the method for liquid chromatography,
Appendix HI D, using the following solutions.
(1) 0.05% w/v of the substance being examined in the mobile
phase
(2) Dilute 1 volume of solution (1) to 50 volumes with the
C. 8-chloro-l l-(piperazin-l-yl)-5H- mobile phase, dilute 5.0 mL of this solution to 100.0 mL
dibenzo [b,e] [1,4] diazepine, with the mobile phase.
1-612 Cocaine Hydrochloride 2016
IDENTIFICATION
Cochineal A. Melting point (see Tests).
DEFINITION B. Composition of fatty acids (see Tests).
Cochineal is the dried female insect, Dactylopim coccus Costa,
containing eggs and larvae. TESTS
M elting point (2.2.14)
CHARACTERISTICS 23 °C to 26 °C.
Odour, characteristic.
Acid value (2.5.2)
Macroscopical Purplish black or purplish grey; about 3.5 to
Maximum 0.5, determined on 20.0 g.
5.5 mm long and 3 to 4.5 mm wide, plano-convex and
somewhat oval in outline; the convex dorsal surface is Peroxide value (2.5.5, Method A)
transversely wrinkled and shows about 11 segments; the flat Maximum 5.0.
or slightly concave ventral surface carries upon the anterior Unsaponifiable m atter (2.5.7)
part two seven-jointed straight antennae, three pairs of short Maximum 1.0 per cent, determined on 5.0 g.
legs, each terminating in a single claw, and a mouth from Alkaline im purities (2.419)
which projects the remains of a long filiform proboscis; these It complies with the test.
appendages are frequendy more or less broken. Easily
Composition of fatty acids (2.422, Method B)
reduced to powder, which is dark red or puce.
Refined coconut oil is melted under gende heating to a
Microscopical Scattered irregularly over the whole dermis are homogeneous liquid prior to sampling.
numerous solitary and grouped, short, tubular wax glands;
Reference solution Dissolve 15.0 mg of tricaproin CRS, 80.0 mg
within each insect are found numerous larvae, which are
of tristearin CRS, 0.150 g of tricaprin CRS, 0.200 g of
characterised by their proboscides appearing as two circular
tricaprylin CRS, 0.450 g of trimyristin CRS and 1.25 g of
coils.
trUaurin CR S in a mixture of 2 volumes of methylene
TESTS chloride R and 8 volumes of heptane R, then dilute to 50 mL
Colour value with the same mixture of solvents heating at 45-50 °C.
To 0.5 g in moderatelyfine powder add 60 mL of phosphate Transfer 2 mL of this mixture to a 10 mL centrifuge tube
buffer p H 8.0 and heat on a water bath for 30 minutes. Cool, with a screw cap and evaporate the solvent in a current of
add sufficient phosphate buffer p H 8.0 to produce 100 mL and nitrogen R. Dissolve with 1 mL of heptane R and 1 mL of
filter. Dilute 5 mL of the filtrate to 100 mL with phosphate dimethyl carbonate R and mix vigorously under gende heating
buffer p H 8.0. The absorbance of the resulting solution at the (50-60 °C). Add, while still warm, 1 mL of a 12 g/L solution
maximum at 530 nm is not less than 0.25, Appendix II B. of sodium R in anhydrous methanol R, prepared with the
Foreign m atter necessary precautions, and mix vigorously for about 5 min.
Complies with the test for foreign matter, Appendix XI D. Add 3 mL of distilled water R and mix vigorously for about
30 s. Centrifuge for 15 min at 1500 g. Inject 1 [iL of the
Water-insoluble m atter
organic phase.
When the insects are placed in water, no insoluble powder
separates. Calculate the percentage content of each fatty add using the
following expression:
Ash
Not more than 7.0%, Appendix XIJ.
Axsc
M icrobial contam ination ^ x’s,c x 100 per cent m/m
1 g is free from Escherichia coli, 10 g is free from Salmonella,
Appendix XVI Bl.
A r r r Is
the corrected peak area of each fatly add in the test
solution:
Rc = 1 for the peaks due to each of the remaining Iodine value (2.5.4, Method A)
specified fatty add methyl esters or any unspecified Maximum 1.0.
fatty add methyl ester. Saponification value (2.5.6)
Composition of thefatty-acidfraction of the oit 160 to 173.
— caproic acid (R& 0.11): maximum 1.5 per cent, Composition of fatty ad ds and fatty alcohols (2.4.22,
— capiyUc acid (R& 0.23): 5.0 per cent to 11.0 per cent, Method C)
— capric acid (Rri 0.56): 4.0 per cent to 9.0 per cent, Use the chromatogram obtained with the following reference
— lauric acid (R& 0.75): 40.0 per cent to 50.0 per cent, solution for identification of the peaks due to the fatty
— myrisdc acid (R& 0.85): 15.0 per cent to 20.0 per cent, alcohols.
— palmitic acid (Rri 0.93): 7.0 per cent to 12.0 per cent,
Reference solution Dissolve the amounts of the substances
— stearic acid (R& 1.00): 1.5 per cent to 5.0 per cent,
listed in the following table in 10 mL of heptane R.
— oleic acid (Rri 1.01): 4.0 per cent to 10.0 per cent,
— linoleic acid (R& 1.03): 1.0 per cent to 3.0 per cent,
— linolenic acid (R& 1.06): maximum 0.2 per cent, Substance Amount (mg)
— arachidic acid (R^ 1.10): maximum 0.2 per cent, Methyl caproate R 10
— eicosenotc add (R& 1.11): maximum 0.2 per cent
Methyl caprylate R 90
Water (2.5.32)
Maximum 0.1 per cent, determined on 1.00 g. Methyl decanoate R 50
Methyl laurate R 20
STORAGE
In a well-filled container, protected from light. Methyl myristate R 10
__________________________________________________________ PhEur Methyl palmitate R 10
Methyl stearate R 10
Decanol R 10
Related substances
Codeine f * \ Liquid chromatography (2.2.29).
* ★
(Ph. Eur. monograph 0076) * Test solution Dissolve 0.100 g of the substance to be
examined and 0.100 g of sodium octanesulfonate R in the
mobile phase and dilute to 10.0 mL with the mobile phase.
Reference solution (a) Dissolve 5.0 mg of codeine
impurity A CRS in the mobile phase and dilute to 5.0 mL
with the mobile phase.
Reference solution (b) Dilute 1.0 mL of reference solution (a)
to 20.0 mL with the mobile phase.
Reference solution (c) Dilute 1.0 mL of the test solution to
C18H21N 03jH20 317.4 6059-47-8 50.0 mL with the mobile phase. Dilute 5.0 mL of this
solution to 100.0 mL with die mobile phase.
Action and use
Reference solution (¿0 To 0.25 mL of the test solution, add
Opioid receptor agonist; analgesic. 2.5 mL of reference solution (a).
PhEur__________________________________________________________ Column:
— size. I = 0.25 m, 0 = 4.6 mm;
DEFINITION
— stationary phase: end-capped octylsQyl silica gel for
7,8-Didehydro-4,5a-epoxy-3-methoxy-17-methylmorphinan- chromatography R (5 Jim).
6a-ol monohydrate.
Mobile phase Dissolve 1.08 g of sodium octanesulfonate R in a
Content mixture of 20 mL of glacial acetic acid R and 250 mL of
99.0 per cent to 101.0 per cent (dried substance). acetonitrile R and dilute to 1000 mL with water R
CHARACTERS Flow rate 2 mL/min.
Appearance Detection Spectrophotometer at 245 nm.
White or almost white, crystalline powder or colourless
Injection 10 (iL.
crystals.
Run time 10 times the retention time of codeine.
Solubility
Relative retention With reference to codeine (retention
Soluble in boiling water, freely soluble in ethanol
time = about 6 min): impurity B = about 0.6;
(96 per cent).
impurity E = about 0.7; impurity A = about 2.0;
IDENTIFICATION impurity C = about 2.3; impurity D = about 3.6.
First identification A , C. System suitability Reference solution (d):
Second identification A , B, D, E — resolution: minimum 3 between the peaks due to codeine
A. Melting point (2.2.14): 155 °C to 159 °C. and impurity A.
B. Ultraviolet and visible absorption spectrophotometry Limits:
(2.2.25). — correction factor,for the calculation of content, multiply die
Test solution To 2.0 mL of solution S (see Tests) add 50 mL
peak area of impurity C by 0.25;
— impurity A: not more than twice the area of the principal
of water R then 10 mL of 1 M sodium hydroxide and dilute to
100.0 mL with water R. peak in the chromatogram obtained with reference
solution (b) (1.0 per cent);
Spectral range 250-350 nm.
— impurities B, C, D, E: for each impurity, not more than
Absorption maximum At 284 nm. twice the area of the principal peak in the chromatogram
Specific absorbance at the absorption maximum About 50 (dried obtained with reference solution (c) (0.2 per cent);
substance). — unspecified impurities: for each impurity, not more than the
C. Infrared absorption spectrophotometry (2.2.24). area of the principal peak in the chromatogram obtained
Preparation Dried substance prepared as a disc of potassium
with reference solution (c) (0.10 per cent);
— sum of impurities other than A: not more than 10 times the
bromide R.
area of the principal peak in the chromatogram obtained
Comparison codeine CRS.
with reference solution (c) (1.0 per cent);
D. To about 10 mg add 1 mL of sulfuric acid R and 0.05 mL — disregard limit. 0.5 times the area of the principal peak in
of ferric chloride solution R2 and heat on a water-bath. A blue the chromatogram obtained with reference solution (c)
colour develops. Add 0.05 mL of nitric acid R. The colour (0.05 per cent).
changes to red.
Loss on drying (2.2.32)
E. It gives the reaction of alkaloids (2.3.1). 4.0 per cent to 6.0 per cent, determined on 1.000 g by
TESTS drying in an oven at 105 °C.
Solution S Sulfated ash (2.414)
Dissolve 50 mg in carbon dioxide-free water R and dilute to Maximum 0.1 per cent, determined on 1.0 g.
10.0 mL with the same solvent.
ASSAY
A ppearance of solution Dissolve 0.250 g in 10 mL of anhydrous acetic acid R
Solution S is clear (2.2.1) and colourless (2.2.2, Method II). Add 20 mL of dioxan R Titrate with 0.1 M perchloric add,
Specific optical rotation (2.2.7) nsing 0.05 mL of crystal violet solution R as indicator.
-142 to -1 4 6 (dried substance). 1 ml. of 0.1 Mperchloric acid is equivalent to 29.94 mg
Dissolve 0.50 g in ethanol (96 per cent) R and dilute to of C18H21N 0 3.
25.0 mL with the same solvent.
2016 Codeine Hydrochloride 1-617
STORAGE
Protected from light.
IMPURITIES
Specified impurities A,B, C, D, E
Other detectable impurities(die following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general E. 7,8-didehydro-4,5a-epoxy-3-methoxy-17-
acceptance criterion for other/unspecified impurities and/or methylmorphinan-6a, 10-diol,
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substancesfor pharmaceutical use): F, G.
F. 7,8-didehydro-4,5a-epoxy-3-methoxy-17-
methylmorphinan-óotj 14-diol,
A. 7,8-didehydro-4,5a-epoxy-3,6a-dimethoxy-17-
methylmorphinan (methylcodeine),
G. 6,7,8,14-tetradehydro-4,5a-epoxy-3,6-dimedioxy-17-
methylmorphinan (thebaine).
PhEur
B. 7,8-didehydro-4,5a-epoxy-17-methylmorphinan-3,6a-diol
(morphine),
***
Codeine Hydrochloride * ★
★ ★
(Codeine Hydrochloride Dihydrate, *****
Ph Eur monograph 1412)
, HCI , 2H20
C. 7,7',8,8/-tetradehydro-4,5a;4,,5 'a-diepoxy-3,3
dimethoxy-17,17 '-dimethyl-2,2 '-bimorphinanyl-6a, 6 'a-diol
(codeine dimer),
C18H22C1N03,2H20 371.9
DEFINITION
7,8-Didehydro-4,5a-epoxy-3-methoxy-17-methylmorphinan-
6a-ol hydrochloride dihydrate.
Content
D. 7,8-didehydro-2-[(7,8-didehydro-4,5a-epoxy-6a-hydroxy- 99.0 per cent to 101.0 per cent (anhydrous substance).
17-methylmorphinan-3-yl)oxy]-4,5a-epoxy-3-methoxy-17-
methylmorphinan-6a-ol (3-0-(codein-2-yl)morphine), CHARACTERS
Appearance
White or almost white, crystalline powder or small, colourless
crystals.
Solubility
Soluble in water, slightly soluble in ethanol (96 per cent),
practically insoluble in cyclohexane.
1-618 Codeine Hydrochloride 2016
Specific optical rotation (2.2.7) potentiometric titration (2.2.20), using 0.1 M sodium
hydroxide. Read the volume added between the 2 points of
-117 t o -121 (anhydrous substance).
inflexion.
Dilute 5.0 mL of solution S to 10.0 mL with water R.
1 mL of 0.1 M sodium hydroxide is equivalent to 33.59 mg
Related substances of C 18H 22CINO3.
Liquid chromatography (2.2.29).
Test solution Dissolve 0.100 g of the substance to be
STORAGE
examined and 0.100 g of sodium octanesulfonate R in the Protected from light.
mobile phase and dilute to 10.0 mL with the mobile phase. IMPURITIES
Reference solution (a) Dissolve 5.0 mg of codeine Spedfied impurities A, B, C, D, E.
impurity A CRS in the mobile phase and dilute to 5.0 mL Other detectable impurities(the following substances would, if
with the mobile phase. present at a sufficient level, be detected by one or other of
Reference sdution (b) Dilute 1.0 mL of reference solution (a) the tests in the monograph. They are limited by the general
to 20.0 mL with the mobile phase. acceptance criterion for other/unspecified impurities and/or
Reference solution (c) Dilute 1.0 mL of the test solution to by the general monograph Substances for pharmaceutical use
50.0 mL with the mobile phase. Dilute 5.0 mL of this (2034). It is therefore not necessary to identify these
solution to 100.0 mL with the mobile phase. impurities for demonstration of compliance. See also 5.10.
Contrd of impurities in substancesfor pharmaceutical use): F, G.
Reference solution (d) To 0.25 mL of the test solution add
2.5 mL of reference solution (a).
Column:
— sizer. I = 0.25 m, 0 = 4.6 mm;
— stationary phase: end-capped octylsüyl silica gelfor
chromatography R (5 |im).
Mobile phase Dissolve 1.08 g of sodium octanesulfonate R in a
mixture of 20 mL of glacial acetic add R and 250 mL of
acetomtrüe R and dilute to 1000 mL with water R.
Flow rate 2 mUmin. A. 7,8-didehydro-4,5a-epoxy-3,6a-dimethoxy-17-
Detection Spectrophotometer at 245 nm. methylmorphinan (methylcodeine),
Injection 10 }iL.
Run time 10 times the retention time of codeine.
Relative retention With reference to codeine (retention
time = about 6 min): impurity B = about 0.6;
2016 Codeine Phosphate 1-619
Codeine Phosphate ★ ★
★ ★
(Codeine Phosphate Hemihydrate, *****
Ph Eur monograph 0074)
B. 7,8-didehydro-4,5a-epoxy-l 7-methylmorphinan-3,6a-diol
(morphine).
by scratching the wall of the tube with a glass rod and Limits:
cooling in iced water. Wash the precipitate with water R and — correction factor, for the calculation of content, multiply the
dry at 100-105 °C. Examine the dried precipitate prepared as peak area of impurity C by 0.25;
discs using potassium bromide R. — impurity A: not more than twice the area of the principal
Comparison Ph. Eur. reference spectrum of codeine. peak in the chromatogram obtained with reference
C. Dissolve 0.20 g in 4 mL of water R. Add 1 mL of a solution (b) (1.0 per cent);
mixture of equal volumes of strong sodium hydroxide solution R — sum of impurities B and E: not more than 4 times the area
and water R and initiate crystallisation, if necessary, by of the principal peak in the chromatogram obtained with
scratching the wall of the tube with a glass rod and cooling in reference solution (c) (0.4 per cent);
iced water. The precipitate, washed with water R and dried at — impurities C, D: for each impurity, not more than twice
100-105 °C, melts (2.2.14) at 155 °C to 159 °C. the area of the principal peak in the chromatogram
obtained with reference solution (c) (0.2 per cent);
D. To about 10 mg add 1 mL of sulfuric acid R and — unspecified impurities: for each impurity, not more than the
0.05 mL of ferric chloride solution R2 and heat on a water- area of the principal peak in the chromatogram obtained
bath. A blue colour develops. Add 0.05 mL of nitric acid R. with reference solution (c) (0.10 per cent);
The colour changes to red. — sum of impurities other than A: not more than 10 times the
E. Loss on drying (see Tests). area of the principal peak in the chromatogram obtained
F. Solution S gives reaction (a) of phosphates (2.3.1). with reference solution (c) (1.0 per cent);
G. It gives the reaction of alkaloids (2.3.1). — disregard limit. 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (c)
TESTS (0.05 per cent).
Solution S
Dissolve 1.00 g in carbon dioxide-free water R prepared from Sulfates (2.413)
distilled water R and dilute to 25.0 ml. with the same solvent.
Maximum 0.1 per cent
Dilute 5 mL of solution S to 20 mL with distilled water R.
pH (2.2.3)
4.0 to 5.0 for solution S. Loss on drying (2.2.32)
1.5 per cent to 3.0 per cent, determined on 1.000 g by
Specific optical rotation (2.2.7)
drying in an oven at 105 °C.
—98 to -102 (dried substance).
Dilute 5.0 mL of solution S to 10.0 mL with water R. ASSAY
Dissolve 0.350 g in a mixture of 10 mL of anhydrous acetic
Related substances
add R and 20 mL of dioxan R. Titrate with 0.1 M perchloric
Liquid chromatography (2.2.29).
acid using 0.05 mL of crystal violet solution R as indicator.
Test solution Dissolve 0.100 g of the substance to be
1 mL of 0.1 M perchloric add is equivalent to 39.74 mg
examined and 0.100 g of sodium octanesulfonate R in the
of C 18H 24NO 7P.
mobile phase and dilute to 10.0 mL with the mobile phase.
Reference solution (a) Dissolve 5.0 mg of codeine STORAGE
impurity A CRS in the mobile phase and dilute to 5.0 mL Protected from light.
with the mobile phase. IMPURITIES
Reference solution (b) Dilute 1.0 mL of reference solution (a) Spedfied impurities A, B, C, D, E
to 20.0 mL with the mobile phase. Other detectable impurities (the following substances would, if
Reference solution (c) Dilute 1.0 mL of the test solution to present at a sufficient level, be detected by one or other of
50.0 mL with the mobile phase. Dilute 5.0 mL of this the tests in the monograph. They are limited by the general
solution to 100.0 mL with the mobile phase. acceptance criterion for other/unspecified impurities and/or
Reference solution (d) To 0.25 mL of the test solution add by the general monograph Substances for pharmaceutical use
2.5 mL of reference solution (a). (2034). It is therefore not necessary to identify these
Column: impurities for demonstration of compliance. See also 5.10.
— size. I = 0.25 m, 0 = 4.6 mm; Control of impurities in substancesfor pharmaceutical use): F, G.
— stationary phase: end-capped octylsUyl silica gelfor
chromatography R (5 jam).
Mobile phase Dissolve 1.08 g of sodium octanesulfonate R in a
mixture of 20 mL of glacial acetic add R and 250 mL of
acetomtrile R and dilute to 1000 mL with water R.
Flow rate 2 ml 7min,
Detection Spectrophotometer at 245 nm.
Injection 10 |iL.
Run time 10 times the retention time of codeine.
A. 7,8-didehydro-4,5a-epoxy-3,6a-dimethoxy-17-
methylmorphinan (methylcodeine),
Relative retention With reference to codeine (retention
time = about 6 min): impurities B and E = about 0.7;
impurity A = about 2.0; impurity C = about 2.3;
impurity D = about 3.6.
System suitability Reference solution (d):
— resolution: minimum 3 between the peaks due to codeine
and impurity A.
2016 Codeine Phosphate Sesquihydrate 1-621
B. 7,8-didehydro-4,5a-epoxy-17-methylmorphinan-3,6a-diol
(morphine).
DEFINITION
7,8-Didehydro-4,5cc-epoxy-3-methoxy-17-methylmorphinan-
6a-ol phosphate sesquihydrate.
Content
98.5 per cent to 101.0 per cent (dried substance).
CHARACTERS
Appearance
White or almost white, crystalline powder or small, colourless
D . 7j8-didehydro-2- [(7,8-didehydro-4,5a-epoxy-6a-hydroxy-
17-methyhnorphinan-3-yl)oxy]-4,5a-epoxy-3-methoxy-l 7- crystals.
methylmorphinan-6a-ol (3-0-(codein-2-yl)morphine), Solubility
Freely soluble in water, slightly soluble in ethanol
(96 per cent).
IDENTIFICATION
First identification B, E, F.
Second identification A ,
C, D, E, F, G
A. Ultraviolet and visible absorption spectrophotometry
(2.2.25).
Test solution Dilute 1.0 mL of solution S (see Tests) to
E. 7,8-didehydro-4,5a-epoxy-3-methoxy-17-
100.0 mL with water R. To 25.0 mL of this solution add
methylmorphinan-ôa, 10-diol,
25 mL of water R then 10 ml. of 1 M sodium hydroxide and
dilute to 100.0 mL with water R.
Spectral range 250-350 nm.
Absorption maximum At 284 nm.
Specific absorbance at the absorption maximum About 38 (dried
substance).
B. Infrared absorption spectrophotometry (2.2.24).
Preparation Dissolve 0.20 g in 4 m l. of water R. Add 1 mL of
F. 7,8-didehydro-4,5a-epoxy-3-methoxy-17- a mixture of equal volumes of strong sodium hydroxide
methylmorphinan-6a, 14-diol, solution R and water R and initiate crystallisation, if necessary,
by scratching the wall of the tube with a glass rod and
cooling in iced water. Wash the precipitate with water R and
dry at 100-105 °C. Examine the dried precipitate prepared as
discs using potassium bromide R.
Comparison Ph. Em. reference spectrum of codeine.
C. Dissolve 0.20 g in 4 mL of water R. Add 1 mL of a
mixture of equal volumes of strong sodium hydroxide solution R
and water R and initiate crystallisation, if necessary, by
G. 6,7,8,14-tetradehydro-4,5a-epoxy-3,6-dimethoxy-l 7- scratching the wall of the tube with a glass rod and cooling in
methylmorphinan (thebaine). iced water. The precipitate, washed with water R and dried at
PhEur 100-105 °C, melts (2.2.14) at 155 °C to 159 °C.
1-622 Codeine Phosphate Sesquihydrate 2016
D. To about 10 mg add 1 mT. of sulfuric add R and 0.05 mL — sum of impurities other than A:not more than 10 times the
of ferric chloride solution R2 and heat on a water-bath. A blue area of the principal peak in the chromatogram obtained
colour develops. Add 0.05 mL of nitric add R. The colour with reference solution (c) (1.0 per cent);
changes to red. — disregard limit: 0.5 times the area of the principal peak in
E. Loss on drying (see Tests). the chromatogram obtained with reference solution (c)
(0.05 per cent).
F. Solution S gives reaction (a) of phosphates (2.3.1).
G. It gives the reaction of alkaloids (2.3.1). Sulfates (2.4.13)
Maximum 0.1 per cent
TESTS Dilute 5 mL of solution S to 20 mL with distilled water R.
Solution S
Dissolve 1.00 g in carbon dioxide-free water R prepared from Loss on drying (2.2.32)
distilled water R and dilute to 25.0 mL with the same solvent.
5.0per cent to 7.5 per cent, determined on 0.500 g by
drying in an oven at 105 °C.
pH (2.2.3)
4.0 to 5.0 for solution S. ASSAY
Dissolve 0.350 g in a mixture of 10 mL of anhydrous acetic
Specific optical rotation (2.2.7)
acid R and 20 mL of dioxan R. Titrate with 0.1 M perchloric
-9 8 to —102 (dried substance).
acid using 0.05 mL of crystal violet solution R as indicator.
Dilute 5.0 mL of solution S to 10.0 mL with water R.
1 mL of 0.1 M perchloric acid is equivalent to 39.74 mg
Related substances of c 18h 24n o 7p .
Liquid chromatography (2.2.29).
STORAGE
Test solution Dissolve 0.100 g of the substance to be
Protected from light.
examined and 0.100 g of sodium octanesulfonate R in the
mobile phase and dilute to 10.0 mL with die mobile phase. IMPURITIES
Reference solution (a) Dissolve 5.0 mg of codeine Speafied impurities A, B, C, D, E
impurity A CRS in the mobile phase and dilute to 5.0 mL Other detectable impurities (the following substances would, if
with the mobile phase. present at a sufficient level, be detected by one or other of
Reference solution (b) Dilute 1.0 mL of reference solution (a) the tests in the monograph. They are limited by the general
to 20.0 mL with the mobile phase. acceptance criterion for other/unspecified impurities and/or
Reference solution (c) Dilute 1.0 mL of the test solution to
by the general monograph Substances for pharmaceutical use
50.0 mL with the mobile phase. Dilute 5.0 mL of this (2034). It is therefore not necessary to identify these
solution to 100.0 mL with the mobile phase. impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): F, G.
Reference solution (d) To 0.25 mL of the test solution add
2.5 mL of reference solution (a).
Column:
— size. I = 0.25 m, 0 = 4.6 mm;
— stationary phase', end-capped octylsHyl silica gd for
chromatography R (5 jam).
Mobile phase Dissolve 1.08 g of sodium octanesulfonate R in a
mixture of 20 mL of glacial acetic add R and 250 mL of
acetomtrUe R and dilute to 1000 mL with water R
Flow rate 2 mL/min. A. 7,8-didehydro-4,5a-epoxy-3,6a-dimethoxy-17-
Detection Spectrophotometer at 245 nm.
methylmorphinan (methylcodeine),
Injection 10 jiL.
Run time 10 times the retention time of codeine.
Relative retention With reference to codeine (retention
time = about 6 min): impurity B = about 0.6;
impurity E = about 0.7; impurity A = about 2.0;
impurity C = about 2.3; impurity D = about 3.6.
System suitability Reference solution (d):
— resolution: minimum 3 between the peaks due to codeine
B. 7,8-didehydro-4,5a-epoxy-17-methylmorphinan-3,6ac-diol
and impurity A.
(morphine),
Limits:
— correction factor, for the calculation of content, multiply die
peak area of impurity C by 0.25;
— impurity A: not more than twice die area of the principal
peak in the chromatogram obtained with reference
solution (b) (1.0 per cent);
— impurities B, C, D, E: for each impurity, not more than
twice the area of the principal peak in the chromatogram
obtained with reference solution (c) (0.2 per cent); h 3c ch3
— unspecified impurities: for each impurity, not more than the
area of the principal peak in the chromatogram obtained C. 7,7 ',8,8 '-tetradehydro-^Sa: 4 ',5 'a-diepoxy-3,3 '-
with reference solution (c) (0.10 per cent); dimethoxy-17,17 '-dimethyl-2,2/-bimorphinanyl-6a,6/a-diol
(codeine dimer),
2016 Codergocrine Mesilate 1-623
Codergocrine Mesilate ★ ★
★ ★
(Ph. Etar. monograph 2060) *****
D . 7,8-<iidehydro-2- [(7,8-didehydro-4,5a-epoxy-6a-hydroxy-
17-methylmorphinan-3-yi) oxy]-4,5a-epoxy-3-methoxy-17-
methylmorphinan-6a-ol (3-0-(codein-2-yl)morphine)J
H3C ^ C H 3
dihydroergocomine
C32 H4 5 N5 O 3 S 660
mesilate
h 3c o
dihydroergocristine
mesilate
C38 H45 N5 O 8 S 708
p ch3
8067-24-1
together with considerations of the quality of starting — stationary phase: octadecylsUyl silica gd for chromatography R
materials, process capability and validation. The general (5 pm).
methods 2.5.37. Methyl, ethyl and isopropyl methanesidfonate in Mobile phase triethylamine R, acetonitrile R, water R
methanesulfonic acid, 2.5.38. Methyl, ethyl and isopropyl (2.5:25:75 V/VfV).
methanesulfonate in active substances and
Flow rate 1.5 mTVmin.
2.5.39. Methanesulfonyl chloride in methanesulfonic add are
Detection Spectrophotometer at 280 nm.
available to assist manufacturers.
Irgection 20 pL.
CHARACTERS
Run time 20 min.
Appearance
White or yellowish powder. Elution order Dihydroergocomine, a-dihydroergocryptine,
dihydroergocristine, J3-dihydroergocryptine.
Solubility
System suitability Test solution:
Sparingly soluble in water, sparingly soluble to soluble in
— resolution: minimum 3 between any 2 consecutive
ethanol (96 per cent), slightly soluble in methylene chloride.
principal peaks.
IDENTIFICATION Composition:
A. Thin-layer chromatography (2.2.27). — dihydroergocomine: 30.0 per cent to 35.0 per cent;
Test solution Dissolve 0.20 g of the substance to be examined — a-dihydroergocryptine: 20.0 per cent to 25.0 per cent;
in a mixture of 1 volume of methanol R and 9 volumes of — dihydroergocristine: 30.0 per cent to 35.0 per cent;
methylene chloride R and dilute to 5 mL with the same — ¡i-dihydroergocryptine. 10.0 per cent to 13.0 per cent;
mixture of solvents. — disregard limit: 1.0 per cent.
Reference solution Dissolve 0.20 g of methanesulfonic acid R in a Related substances
mixture of 1 volume of methanol R and 9 volumes of Thin-layer chromatography (2.2.27). Perform the test as rapidly
methylene chloride R and dilute to 5 mL with the same as possible and protectedfrom direct light Prepare the test solution
mixture of solvents. last and immediately before application on the plate.
Plate TLC silica gd plate R. Test solution Dissolve 0.40 g of the substance to be examined
Mobile phase water R, concentrated ammonia R, butanol R, in a mixture of 1 volume of methanol R and 9 volumes of
acetone R (5:10:20:65 VIVIVIV). methylene chloride R and dilute to 5.0 mL with the same
Application 10 |iL. mixture of solvents.
Development Over 2/3 of the plate. Reference solution (a) Dissolve 40 mg of dihydroergocristine
mesilate CRS in a mixture of 1 volume of methanol R and
Drying In a current of cold air for not more than 1 min.
9 volumes of methylene chloride R and dilute to 10.0 mL with
Detection Spray with a 1 g/L solution of bromocresol purple R the same mixture of solvents. Dilute 3.0 mL of the solution
in methanol R, adjusted to a violet-red colour with 0.05 mL to 50.0 mL with a mixture of 1 volume of methanol R and
of dilute ammonia R l. 9 volumes of methylene chloride R.
Drying In a current of hot air at 100 °C. Reference solution (b) To 2.0 mL of reference solution (a),
Results The principal spot in the chromatogram obtained with add 1.0 mL of a mixture of 1 volume of methanol R and
the test solution is similar in position and colour to the 9 volumes of methylene chloride R.
principal spot in the chromatogram obtained with the Reference solution (c) To 1.0 mL of reference solution (a), add
reference solution. 2.0 mL of a mixture of 1 volume of methanol R and
B. Examine the chromatograms obtained in the test for 9 volumes of methylene chloride R.
composition. Reference solution (d) To 1.0 mL of reference solution (a),
Results The 4 principal peaks in the chromatogram obtained add 5.0 mL of a mixture of 1 volume of methanol R and
with the test solution are similar in retention time to the 9 volumes of methylene chloride R.
4 principal peaks in the chromatogram obtained with the Plate T LC silica gel plate R.
reference solution.
Mobile phase concentrated ammonia R, methanol R, ethyl
TESTS acetate R, methylene chloride R (1:3:50:50 VIVIVIV).
pH (2.2.3) Application 10 |iL.
4.2 to 5.2.
Drying In the dark for 2 min after the application of the last
Dissolve 0.10 g in carbon dioxide-free water R and dilute to solution.
20 mL with the same solvent.
First development In an unsaturated tank, over 2/3 of the
Composition plate.
Liquid chromatography (2.2.29): use the normalisation Drying In a current of cold air for not more than 1 min.
procedure.
Second development In an unsaturated tank, over 2/3 of the
Test solution Dissolve 20 mg of the substance to be examined
plate; use freshly prepared mobile phase.
in a mixture of 1 volume of anhydrous ethanol R and
Drying In a current of cold air for not more than 1 min.
2 volumes of a 10 g/L solution of tartaric add R and dilute to
10 mL with the same mixture of solvents. Detection Spray thoroughly with dimethylaminobenzaldehyde
solution R7 and dry in a current of hot air until die spot in
Reference solution Dissolve 20 mg of codergocrine mesilate CRS
the chromatogram obtained with reference solution (d) is
in a mixture of 1 volume of anhydrous ethanol R and
2 volumes of a 10 g/L solution of tartaric add R and dilute to clearly visible.
10 mL with the same mixture of solvents. System suitability Test solution:
Column:
— the chromatogram shows at least 3 separated secondary
— size. I ~ 0.15 m, 0 = 4.6 mm; spots.
2016 Cod-liver Oil 1-625
173.4 173.2 173.0 172.8 172.6 172.4 172.2 172.0 171.8 ppm
System suitability: 20:4 n-3, 20:5 n-3, 22:1 n-11, 22:1 n-9, 21:5 n-3, 22:5 n-3,
— signal-to-noise ratio:minimum 5 for the smallest relevant 22:6 n-3).
peak corresponding to a C 18:4 signal (in the Linoleic acid (2.4.29)
range 8 172.95-172.99 ppm); 3.0 per cent to 11.0 per cent.
— peak width at half-height, maximum 0.02 ppm for the
central CDCI3 signal (at 5 77.16 ppm). ASSAY
EPA and DHA (2.4.29)
Calculation of positional distribution ((¡(2)-acyl) Use the
See the chromatogram shown in Figure 2398.-2.
following expression:
Vitamin A
100 x /3 Carry out the test as rapidly as possible, avoiding exposure to
actinic light and air, oxidising agents, oxidation catalysts (for
a+ 0
example, copper and iron) and adds.
a = peak area of the corresponding a-carbonyl peak; Use method A. If method A is found not to be valid, use
/? = peak area of {$-carbonyl peak from C22:6 n-3, method B.
C20:5 n-3 or C l8:4 n-3, respectively. M ETHOD A
Limits: Ultraviolet absorption spectrophotometry (2.2.25).
— positional distribution (fi(2)-acyl): Test solution To 1.00 g in a round-bottomed flask, add 3 mL
— cervomc (docosahexaenoic) acid (C22:6 n-3; DHA): of a freshly prepared 50 per cent m/m solution of potassium
71 per cent to 81 per cent; hydroxide R and 30 mL of anhydrous ethanol R. Boil under
— txmnodomc (eicosapentaenoic) acid (C20:5 n-3 EPA): reflux in a current of nitrogen R for 30 min. Cool rapidly and
32 per cent to 40 per cent; add 30 mL of water R. Extract with 50 mL of ether R. Repeat
— morocdc add ( C l8:4 n-3): 28 per cent to 38 per cent. the extraction 3 times and discard the lower layer after
Composition of fatty acids (2.4.29) complete separation. Wash the combined upper layers with
For identification of the peaks, see the chromatogram shown 4 quantities, each of 50 mT, of water R, and evaporate to
in Figure 2398.-2. dryness under a gentle current of nitrogen R at a temperature
not exceeding 30 °C or in a rotary evaporator at a
The 24 largest peaks of the methyl esters account for more
temperature not exceeding 30 °C under reduced pressure
than 90 per cent of the total area (these correspond to, in
(water ejector). Dissolve the residue in sufficient 2-
common elution order 14:0, 15:0, 16:0, 16:1 n-7, 16:4 n-1,
propanol R1 to give an expected concentration of vitamin A
18:0, 18:1 n-9, 18:1 n-7, 18:2 n-6 , 18:3 n-3, 18:4 n-3,
equivalent to 10-15 IU/mL.
20:1 n-11, 20:1 n-9, 20:1 n-7, 20:2 n-6 , 20:4 n-6 , 20:3 n-3,
2016 Cod-liver Oil 1-627
24
19
? 9
13
1011 20 23
12
22
'iV- n W i nAv-n/ X A-
mi|Tiii|iiii|iiii|iiiiii|n]'i[iinjiiii|iiii|iiii|mi|ii!i|in
8 10 12 14 16 18 20 22 24 26 min
1. C14:0 5. C16:4 n-1 9. C18:2 n-6 13. C20;l n-9 17. C20:3 n-3 21. C22:l n-9
2. C15:0 6. C18:0 10 C18:3 n-3 14. C20:l n-7 18. C20:4 n-3 22. C21:5 n-3
3. C16:0 7. C18:l n-9 11. C18:4 n-3 15. C20:2 n-6 19. C20:5 n-3 23. C22:5 n-3
4. C16:l n-7 8. C18:l n-7 12. C20:l tt-11 16. C20:4 n-6 20. C22:l n-11 24. C22:6 n-3
Figure 2398.-2. - Chromatogram for the test for composition of fatty acids of farmed cod-liver oil
Measure the absorbances of the solution at 300 nm, 310 nm, A Designates the absorbance at the wavelength indicated by
325 nm and 334 nm and at the wavelength of maximum the subscript
absorption with a suitable spectrophotometer in specially If ^325 has a value greater than A325jCon: /0.970, calculate the
matched 1 cm cells, using 2-propanol R1 as the compensation content of vitamin A using the following expression:
liquid.
Calculate the content of vitam in A, as all-irans-retinol, in
4 1 8 2 1
International Units per gram, using the following expression: ^325, corr X _ XV
100m
1821
¿325 x — T/
— x V The assay is not valid unless:
100 m — the wavelength of maximum absorption lies between
323 nm and 327 nm;
■^325 = absorbance at 325 nm; — the absorbance at 300 nm relative to that at 325 nm is at
m = mass of the substance to be examined, in most 0.73.
grams;
= total volume of solution containing 10-15 IU of M ETH O D B
vitamin A per millilitre; Liquid chromatography (2.2.29).
1821 = conversion factor for the specific absorbance of Test solution Prepare duplicates. To 2.00 g in a round-
all-rranî-retinol, in International Units. bottomed flask, add 5 mL of a freshly prepared 100 g/L
solution of ascorbic acid R, 10 mL of a freshly prepared
The above expression can be used only if ^325 has a value
800 g/L solution of potassium hydroxide R and 100 mL of
not greater than -^325,con- /0.970, where A 32^,can is the
anhydrous ethanol R. Boil under a reflux condenser on a
corrected absorbance at 325 nm and is given by the following
water-bath for 15 min. Add 100 mL of a 10 g/L solution of
equation:
sodium chloride R and cool. Transfer the solution to a 500 mL
separating funnel, rinsing the round-bottomed flask with
¿■325, corr = 6.815^-325 —.2.555^310 -r’ 4.260^334 about 75 mL of a 10 g/L solution of sodium chloride R and
1-628 Cod-liver Oil 2016
pressure (water ejector) and fill with nitrogen R when A1 = area (or height) of the peak due to cholecalciferol
evaporation is completed. Alternatively, evaporate the solvent in the chromatogram obtained with test solution
under a gentle current of nitrogen R at a temperature not (a);
exceeding 30 °C. Dissolve the residue in 1.5 mL of the A2 = area (or height) of the peak due to cholecaldferol
mobile phase described under Purification. Gentle heating in in the chromatogram obtained with test solution
an ultrasonic bath may be required. A largefraction of the (b);
white residue is cholesterol, constituting approximately 50 per cent A3 = area (or hdght) of the peak due to ergocalciferol
mJm of the unsaponifiable matter of cod4ruer oH in the chromatogram obtained with reference
Test solution (b) Prepare duplicates. To 4.00 g add 2.0 mL of solution (b);
the internal standard solution and proceed as described for A4 = area (or hdght) of the peak due to ergocalciferol
test solution (a). in the chromatogram obtained with test solution
Reference solution (a) Dissolve 0.50 mg of cholecalciferol CRS (b);
in 100.0 mL of anhydrous ethanol R A5 = area (or hdght) of a possible peak in the
chromatogram obtained with test "Solution (a) with
Reference solution (b) In a round-bottomed flask, add 2.0 mL
the same retention time as the peak co-eluting
of reference solution (a) and 2.0 mL of the internal standard
with ergocalciferol in test solution (b);
solution and proceed as described for test solution (a).
A6 = area (or hdght) of the peak due to cholecalciferol
PURIFICATION in the chromatogram obtained with reference
Column: solution (b);
— sizer. I = 0.25 m, 0 = 4.6 mm; Vx — total volume of reference solution (a) (100 mL);
— stationary phase: nitrile silica gd for chromatography R V2 = volume of reference solution (a) used for
(10 pm). preparing reference solution (b) (2.0 mL).
Mobile phase isoamyl alcohol R, hexane R (1.6:98.4 VIV). STORAGE
Flow rate 1.1 mlVmin. In an airtight and well-filled container, protected from light
Detection Spectrophotometer at 265 nm. If no antioxidant is added, store under an inert gas.
Injection 350 pL of reference solution (b) and test Once the container has been opened, its contents are used as
solutions (a) and (b). Collect each ehiate from 2 min before soon as possible and any part of the contents not used at
until 2 min after the retention time of cholecalciferol, in a once is protected by an atmosphere of inert gas.
ground-glass-stoppered tube containing 1 mL of a 1 g/L LABELLING
solution of butylhydroxytoluene R in hexane R. Evaporate
The labd states:
separately to dryness at a temperature not exceeding 30 °C
— the concentration of EPA and DHA as a sum;
under a gentle current of nitrogen R Dissolve each residue in — the number of International Units of vitamin A per gram;
1.5 mL of acetomtrUe R.
— the number of International Units of vitamin D3 per
DETERMINATION gram.
Column: PhEur
— sizer. I = 0.15 m, 0 = 4.6 mm;
— stationary phase: octadecylsUyl silica gd for chromatography R
(5 pm).
Mobile phase phosphoric add R, 96 per cent VIV solution of
acetonitrUe R (0.2:99.8 VIV). Cod-liver Oil (Type A) ★ ★
★ ★
Flow rate 1.0 mlVmin. (Ph. Eur. monograph 1192) *****
Detection Spectrophotometer at 265 nm.
Injection 2 quantities not exceeding 200 pL of each of the Action and use
3 solutions obtained under Purification. Source of vitamins A and D.
System suitability: Each IU of vitamin D3 is equivalent to 0.025 pg of
— resolution: minimum 1.4 between the peaks due to colecalciferol.
ergocalciferol and cholecalciferol in the chromatogram
PhEtr.
obtained with reference solution (b);
— the results obtained with the test solution (b) duplicates DEFINITION
do not differ by more than 5 per cent. Purified fatty oil obtained from the fresh livers of wild cod,
Calculate the content of vitamin D3 in International Units Gadus morhua L. and other spedes of Gadidae, solid
per gram using the following expression, taking into account substances being removed by cooling and filtering. A suitable
the assigned content of cholecaldfervl CRS: antioxidant may be added.
Content
Ai Az 7712 Vi — vitamin A: 600 IU (180 pg) to 2500 IU (750 pg) per
X ----- X — x 40 gram;
Ae As mi Vi
A4 - x A% — vitamin D 3: 60 IU (1.5 pg) to 250 IU (6.25 pg) per gram.
Ai
PRODUCTION
mi = mass of the sample in test solution (b), in grams; The content of dioxins and dioxin-like PCBs
m2 = total mass of cholecaldferol CRS used for the (polychlorinated biphenyls) is controlled using methods and
preparation of reference solution (a), in limits in accordance with the requirements set in the
European Union or other applicable regulations.
micrograms (500 pg);
1-630 Cod-liver Oil (Type A) 2016
55 - 75 225
Detector 280
2016 Cod-liver Oil (Type A) 1-631
Figure 1192.-1. - Chromatogram for the testfor composition offatty acids of cod-liver oil (type A)
Detection Flame ionisation. Use method A. If method A is found not to be valid, use
Injection 1 |iL, twice. method B.
System suitability: METHOD A
— the 15 fatty adds to be tested are satisfactorily identified Ultraviolet absorption spectrophotometry (2.2.25).
from the chromatogram shown in Figure 1192.-1; Test solution To 1.00 g in a round-bottomed flask, add 3 mL
— injection of a mixture of equal amounts of methyl of a freshly prepared 50 per cent m/m solution of potassium
palmitate R, methyl stearate R, methyl arachidate R and
hydroxide R and 30 mL of anhydrous ethanol R. Boil under
methyl behenate R gives area percentages of 24.4, 24.8,
reflux in a current of nitrogen R for 30 min. Cool rapidly and
25.2and 25.6 (± 0.5 per cent), respectively; add 30 mL of water R. Extract with 50 mL of ether R. Repeat
— resolution: minimum 1.3 between the peaks due to methyl the extraction 3 times and discard the lower layer after
oleate and methyl os-vaccenate; the resolution between complete separation. Wash the combined upper layers with
the pair due to methyl gadoleate and methyl gondoate is 4 quantities, each of 50 mL, of water R, and evaporate to
sufficient for purposes of identification and area dryness under a gentle current of nitrogen R a t a temperature
measurement. not exceeding 30 °c or in a rotary evaporator at a
Calculate the area per cent for each fatty add methyl ester temperature not exceeding 30 °c under reduced pressure
using the following expression: (water ejector). Dissolve the residue in suffirient
2-propanol R1 to give an expected concentration of vitamin A
equivalent to 10-15 IU/mL.
x 100
At Measure the absorbances of the solution at 300 nm, 310 nm,
325 nm and 334 nm and at the wavelength of maximum
Ax = peak area of fatty add x; absorption with a suitable spectrophotometer in spedally
At = sum of the peak areas (up to C22:6 n-3). matched 1 cm cells, using 2-propanol R1 as the compensation
liquid.
The calculation is not valid unless:
— the total area is based only on peaks due solely to fatty Calculate thê content of vitamin A, as all-iTCOTj-retinol, in
add methyl esters; International Units per gram, using the following expression:
— the number of fatty add methyl ester peaks exceeding
0.05 per cent of the total area is at least 24; ¿325 X 1821 X V
T/
— the 24 largest peaks of the methyl esters account for more 100m
than 90 per cent of the total area (these correspond to, in
common elution order 14:0, 15:0, 16:0,16:1 n-7, A-Ì25 = absorbance at 325 nm;
16:4 n-1, 18:0, 18:1 n-9, 18:1 n-7, 18:2 n-6, 18:3 n-3, m = mass of the substance to be examined, in grams;
18:4 n-3, 20:1 n-11, 20:1 n-9, 20:1 n-7, 20:2 n-6, 20:4 V = total volume of solution containing 10-15 IU of
n-6, 20:3 n-3, 20:4 n-3, 20:5 n-3, 22:1 n-11, 22:1 n-9, vitamin A per millilitre;
21:5 n-3, 22:5 n-3, 22:6 n-3). 1821 = conversion factor for the specific absorbance of all-
mwis-retinol, in International Units.
ASSAY
V ita m in A The above expression can be used only if Aws has a value
Carry out the test as rapidly as possible, avoiding exposure to not greater than A^2ĩfifxJ0.91ồ, where A 325 corr is the
actinic light and air, oxidising agents, oxidation catalysts (for corrected absorbance at 325 nm and is given by the following
example, copper and iron) and acids. equation:
1-632 Cod-liver Oil (Type A) 2016
red primary solution add 25.0 mL of yellow primary solution tightly, mix and heat on a water-bath for 30 min. Cool to
and dilute to 50.0 mL with a 10 g/L solution of hydrochloric 40-50 °C, add 1 mL of trimethylpentane R, cap and vortex or
acid R (2.2.2, Method II). shake vigorously for at least 30 s. Immediately add 5 mL of
saturated sodium chloride solution R, cover with nitrogen R, cap
Relative density (2.2.5) and vortex or shake thoroughly for at least 15 s. Allow the
0.917 to 0.930.
upper layer to become clear and transfer to a separate tube.
Refractive index (2.2.6) Shake the methanol layer once more with 1 mL of
1.477 to 1.484. trimethylpentane R and combine the trimethylpentane extracts.
Acid value (2.5.1) Wash the combined extracts with 2 quantities, each of 1 mL,
Maximum 2.0. of water R and dry over anhydrous sodium sulfate R. Prepare
2 solutions for each sample.
2016 Cod-liver Oü (Type B) 1-635
Figure 1193.-1. - Chromatogram for the testfor composition offatty adds of cod-liver oU (type B)
Calculate the content of vitamin A, as all-mzm-retinol, in Reference solution (a) Prepare a solution of retinol acetate CRS
International Units per gram using the following expression: in 2-propanol R l so that 1 mL contains about 1000 IU of
all-mzm-retinol.
ü 1821 Tr The exact concentration of reference solution (a) is assessed
^ X M0SXV by ultraviolet absorption spectrophotometry (2.2.25). Dilute
reference solution (a) with 2-propanol R l to a presumed
■^325 = absorbance at 325 nm; concentration of 10-15 IU/mL and measure the absorbance
m = mass of the substance to be examined, in at 326 nm in matched 1 cm cells using 2-propanol R l as the
grams; compensation liquid.
V total volume of solution containing 10-15 IU of Calculate the content of vitamin A in International Units per
vitamin A per millilitre; millilitre of reference solution (a) using the following
1821 conversion factor for the specific absorbance of expression, taking into account the assigned content of retinol
all-mzm-retinol, in International Units. acetate CRS:
The above expression can be used only if A 325 has a value
not greater than A 32s,con/0.970 where A 325,COn is the . 1900 x V2
corrected absorbance at 325 nm and is given by the 336 X 100 x Vi
equation:
-^326 = absorbance at 326 nm;
■^325,corr = 6 . 8 I 5 .A325 — 2 .5 5 5 ^ 3 1 0 — 4 .2 6 0 ^ 3 3 4 Vx = volume of reference solution (a) used;
V2 = volume of the diluted solution;
A Designates the absorbance at the wavelength indicated by 1900 = conversion factor for the specific absorbance of
the subscript. retinol acetate CRS, in International Units.
If A 325 has a value greater than A 325jCOJ0.970, calculate the Reference solution (b) Proceed as described for the test
content of vitamin A using the following expression: solution but using 2.00 mL of reference solution (a) in place
of the substance to be examined.
The exact concentration of reference solution (b) is assessed
■^325,coit X 1821 X 1V/ by ultraviolet absorption spectrophotometry (2.2.25). Dilute
100m
reference solution (b) with 2-propanol R l to a presumed
The assay is not valid unless: concentration of 10-15 IU/mL of all-mzm-retinol and
— the wavelength of maximum absorption lies between measure the absorbance at 325 nm in matched 1 cm cells
323 nm and 327 nm; using 2-propanol R l as the compensation liquid.
— the absorbance at 300 nm relative to that at 325 nm is at Calculate the content of all-mzm-retinol in International
most 0.73. Units per millilitre of reference solution (b) from the
M ETHODE expression:
liquid chromatography (2.2.29).
. 1821 x V3
Test solution Prepare duplicates. To 2.00 g in a round-
325 X 100 x VA
bottomed flask, add 5 mL of a freshly prepared 100 g/L
solution of ascorbic acid. R and 10 mL of a freshly prepared A 325 = absorbance at 325 nm;
800 g/L solution of potassium hydroxide R and 100 mL of V3 = volume of the diluted solution;
anhydrous ethanol R. Boil under a reflux condenser on a
F4 = volume of reference solution (b) used;
water-bath for 15 min. Add 100 mL of a 10 g/L solution of 1821 = conversion factor for the specific absorbance of
sodium chloride R and cool. Transfer the solution to a 500 mL
aU-mzm-retinol, in International Units.
separating funnel, rinsing the round-bottomed flask with
about 75 mL of a 10 g/L solution of sodium chloride R and Column:
then with 150 mL of a mixture of equal volumes of ether R — size. I = 0.25 m, 0 = 4.6 mm;
and light petroleum R l. Shake for 1 min. When the layers — stationary phase, octadecylsifyl silica gelfor chromatography R
have separated completely, discard the lower layer and wash (5-10 pm).
the upper layer, first with 50 mL of a 30 g/L solution of Mobile phase water R, methanol R (3:97 VIV).
potassium hydroxide R in a 10 per cent VIV solution of Flow rate 1 mlVmin.
anhydrous ethanol R and then with 3 quantities, each of Detection Spectrophotometer at 325 nm.
50 mL, of a 10 g/L solution of sodium chloride R. Filter the
Injection 10 pL; inject in triplicate the test solution and
upper layer through 5 g of anhydrous sodium sulfate R on a
reference solution (b).
fast filter paper into a 250 mL flask suitable for a rotary
evaporator. Wash the funnel with 10 mL of fresh extraction Retention time All-mzm-retinol = 5 + 1 min.
mixture, filter and combine the upper layers. Distil them at a System suitability:
temperature not exceeding 30 °C under reduced pressure — the chromatogram obtained with the test solution shows a
(water ejector) and fill with nitrogen R when evaporation is peak corresponding to the peak due to all-mzm-retinol in
completed. Alternatively evaporate the solvent under a gende the chromatogram obtained with reference solution (b);
current of nitrogen R at a temperature not exceeding 30 °C. — the results obtained with the duplicate test solutions do
Dissolve the residue in 2-propanol R, transfer to a 25 mL not differ by more than 5 per cent;
volumetric flask and dilute to 25 mL with 2-propanol R. — the recovery of all-mzm-retinol in reference solution (b) as
Gentle heating in an ultrasonic bath may be required. A large assessed by direct absorption spectrophotometry is greater
fraction of the white residue is cholesterol, constituting than 95 per cent
approximately 50 per cent m/m of the unsaponifiable matter of Calculate the content of vitamin A using the following
cod-liver criL expression:
2016 Cod-liver Oil (Type B) 1-637
Limits:
— impurity A: not more than 3.5 times the area of the
principal peak in the chromatogram obtained with
reference solution (b) (3.5 per cent);
— any other impurity: not more than the area of the principal
peak in the chromatogram obtained with reference
solution (b) (1 per cent);
— total: not more than 5 times the area of the principal peak
in the chromatogram obtained with reference solution (b) C. N-[(75'37bi?,10a5)-l,2,3,9-tetramethoxy-8-oxo-
(5 per cent); 5,6 ,7,7b,8,10a-
— disregard limit, the area of the principal peak in the hexahydrobenzo [a] cyclopenta [3,4] cyclobuta [1,2-
chromatogram obtained with reference solution (c) c]cyclohepten-7-yl]acetamide (P-lumicolchicine),
(0.05 per cent).
Colchiceine
Maximum 0.2 per cent.
Dissolve 50 mg in water R and dilute to 5 mL with the same
solvent. Add 0.1 mL of ferric chloride solution R l.
The solution is not more intensely coloured than a mixture
of 1 m l. of red primary solution, 2 mL of yellow primaiy
solution and 2 mL of blue primary solution (2.2.2,
Method II).
Chloroform (2.4.24)
D. N - [(7S, 12ai?J-3-(P-D-glucopyranosyloxy)-l,2,10-
Maximum 500 ppm.
trimethoxy-9-oxo-5,6,7,9-tetrahydrobenzo[a]heptalen-7-
Ethyl acetate (2.4.24) yl]acetamide (colchicoside),
Maximum 6.0 per cent m/m.
W ater (2.5.12)
Maximum 2.0 per cent, determined on 0.500 g.
Sulfa ted ash (2.4.14)
Maximum 0.1 per cent, determined on 0.5 g.
ASSAY
Dissolve 0.250 g with gentle heating in a mixture of 10 mL
of acetic anhydride R and 20 mL of toluene R. Titrate with
0.1 M perchloric add, determining the end-point E. N-[(7S, 12a2?a)-3-hydroxy-l,2,10-trimethoxy-9-oxo-
potentiometrically (2.2.20). 5,6,7,9-tetrahydrobenzo [a] heptalen-7-yl] acetamide (3-0-
1 mL of 0.1 M perchloric acid is equivalent to 39.94 mg of demethylcolchicine),
C 22H 25NO 6.
STORAGE
Protected from light.
IMPURITIES
A. N - [(75,12ai?J-l ,2,3,10-tetramethoxy-9-oxo-5,6,7,9-
tetrahydrobenzo [a]heptalen-7-yl]formamide (iV-deacetyl-N-
formylcolchicine),
OCH3
B. (-)-N-[(7S, 12 a 5 J-l,2,3,10-tetramethoxy-9-oxo-5,6,7,9-
tetrahydrobenzo[a]heptalen-7-yl]acetamide (conformational
isomer),
1-640 Colecalciferol 2016
Paediatric Vitamins A, C and D Oral Drops Relative retention With reference to cholecalciferol (retention
time = about 19 min): pre-cholecalciferol = about 0.5;
When cholecalciferol or vitamin D3 is prescribed or
impurity A = about 0.6.
demanded, Colecalciferol shall be dispensed or supplied.
System suitability: reference solution (b):
When calciferol or vitamin D is prescribed or demanded,
Colecalciferol or Ergocalciferol shall be dispensed or — resolution: m in im u m 1.5 between the peaks due to pre-
supplied. cholecalciferol and impurity A.
Limits:
PhEur___________________________________________________________ — impurity A: not more than the area of the principal peak
DEFINITION in the chromatogram obtained with reference solution (c)
(5Z,7£)-9,10-Secocholesta-5,7,10(19)-trien-3|3-ol. (0.1 per cent);
— unspecified impurities: for each impurity, not more than the
Content area of the principal peak in the chromatogram obtained
97.0 per cent to 102.0 per cent. with reference solution (c) (0.10 per cent);
A reversible isomerisation to pre-cholecalciferol takes place in — total: not more than 10 times the area of the principal
solution, depending on temperature and time. The activity is peak in the chromatogram obtained with reference
due to both compounds (see Assay). solution (c) ( 1.0 per cent);
1 mg of cholecalciferol is equivalent to 40 000 IU of — disregard limit. 0.5 times the area of the principal peak in
antirachitic activity (vitamin D) in rats. the chromatogram obtained with reference solution (c)
(0.05 per cent); disregard the peak due to pre-
CHARACTERS
cholecalciferol.
Appearance
White or almost white crystals. ASSAY
Solubility Liquid chromatography (2.2.29) as described in the test for
Practically insoluble in water, freely soluble in ethanol related substances with the following modification.
(96 per cent), soluble in trimethylpentane and in fatty oils. Injection Test solution and reference solution (a).
It is sensitive to air, heat and light. Solutions in solvents Calculate the percentage content of C27H 44O taking into
without an antioxidant are unstable and are to be used account the assigned content of cholecalciferol CRS and, if
immediately. necessary, the peak due to pre-cholecalciferol.
IDENTIFICATION STORAGE
Infrared absorption spectrophotometry (2.2.24). Under nitrogen, in an airtight container, protected from light,
Comparison cholecalciferol CRS. at a temperature of 2 °C to 8 °C.
The contents of an opened container are to be used
TESTS
immediately.
Specific optical rotation (2.2.7)
+ 105 to 4- 112, determined within 30 min of preparing the IMPURITIES
solution. Specified impurities A
Dissolve 0.200 g rapidly in aldehyde-free alcohol R without Other detectable impurities (the following substances would, if
heating and dilute to 25.0 mL with the same solvent. present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by die general
Related substances
acceptance criterion for other/unspecified impurities and/or
Liquid chromatography (2.2.29). Prepare the solutions
by the general monograph Substances for pharmaceutical use
immediately before use, avoiding exposure to actinic light and air.
2016 Colecalciferol 1-641
DEFINITION
Solution of Cholecalciferol (0072) in a suitable vegetable fatty
oil, authorised by the competent authority.
Content
90.0 per cent to 110.0 per cent of the cholecalciferol content
A. (5E,7E)-9, 1O-secocholesta-5,7,10(19)-trien-3^-ol (tram- stated on the label, which is not less than 500 000 IU/g.
cholecalciferol, mmi-vitamin D3), It may contain suitable stabilisers such as antioxidants.
CHARACTERS
Appearance
Clear, yellow liquid.
Solubility
Practically insoluble in water, slightly soluble in anhydrous
ethanol, miscible with solvents of fats.
Partial solidification may occur, depending on the
temperature.
B. cholesta-5j7-dien-3{J-ol (7,8-didehydrocholesterol,
provitamin D3), IDENTIFICATION
First identificadon A, C
Second identification A, B
A. Thin-layer chromatography (2.2.27). Prepare the solutions
immediately before use.
Test solution Dissolve an amount of the preparation to be
examined corresponding to 400 000 IU in ethylene chloride R
containing 10 g/L of squalane R and 0.1 g/L of
butylhydroxytoluene R and dilute to 4 mL with the same
C. 9 p, 10a-cholesta-5j7-dien-3^-ol (himisterol3), solution.
Reference solution (a) Dissolve 10 mg of cholecalciferol CRS in
ethylene chloride R containing 10 g/L of squalane R and
0.1 g/L of butylhydroxytoluene R and dilute to 4 mL with the
same solution.
Reference solution (b) Dissolve 10 mg of ergocalciferol CRS in
ethylene chloride R containing 10 g/L of squalane R and
0.1 g/L of bvaylhydroxytohiene R and dilute to 4 mL with the
same solution.
Plate TLC silica gel G plate R.
Mobile phase A 0.1 g/L solution of butylhydroxytoluene R in a
mixture of equal volumes of cydohexane R and peroxide-free
ether R.
D. (6E)-9,10-secocholesta-5(1 0)36j8(14)-trien-3f}-ol
(iso-tachysterol3), Application 20 |iL.
Development Immediately, protected from light, over a path of
15 cm.
Drying in air.
Detection Spray with sulfuric acid R.
Results The chromatogram obtained with the test solution
shows immediately a bright yellow principal spot which
rapidly becomes orange-brown, then gradually greenish-grey,
rem ain in g so for 10 min. This spot is similar in position,
colour and size to the spot in the chromatogram obtained
with reference solution (a). The chromatogram obtained with
reference solution (b) shows immediately at the same level an
E. (6E)-9,10-secocholesta-5(10),6,8-trien-3P-ol orange principal spot which gradually becomes reddish-
(tachysterol3). brown and remains so for 10 min.
PhEur
1-642 Colecalciferol 2016
B. Ultraviolet and visible absorption spectrophotometry System suitability', reference solution (b):
(2.2.25). — resolution', m in im u m 1.0 between the
peaks due to pre-
Test solution Prepare a solution in cycbhexane R containing cholecaltiferol and mzw-cholecalciferol; if necessary adjust
the equivalent of about 400 IU/mL. the proportions of the constituents and the flow rate of
the mobile phase to obtain this resolution;
Spectral range 250-300 nm.
— repeatability: maximum relative standard deviation of
Absorption maximum At 267 nm. 1.0 per cent for the peak due to cholecalciferol after
C. Examine the chromatograms obtained in the assay. 6 injections.
Results The principal peak in the chromatogram obtained Calculate the conversion factor if) using the following
with the test solution is similar in retention time to the expression:
principal peak in the chromatogram obtained with reference
solution (a). K -L
TESTS M
A d d value (2.5./)
Maximum 2.0. K area (or height) of the peak due to cholecalciferol
Dissolve 5.0 g in 25 mL of the prescribed mixture of in the chromatogram obtained with reference
solvents. solution (d);
Peroxide value (2.5.5, Method A) = area (or height) of the peak due to cholecalciferol
Maximum 20. in the chromatogram obtained with reference
solution (e);
Related substances
M = area (or height) of the peak due to pre-
The thresholds indicated under Related substances
cholecalciferol in the chromatogram obtained with
(Table 2034.-1) in the general monograph Substances for
reference solution (e).
pharmaceutical use (2034) do not apply.
The value of/ d eterm ined in duplicate on different days may
ASSAY
be used during the entire procedure.
Carry out the assay as rapidly as possible, avoiding exposure to
Calculate the content of cholecalciferol in International Units
actinic light and air.
per gram using the following expression:
Liquid chromatography (2.2.29).
Test solution Dissolve a quantity of the preparation to be
m V SD + ( f x Sp)
examined, weighed with an accuracy of 0.1 per cent, X — X x 40 000 x 1000
equivalent to about 400 000 IU, in 10.0 mL of toluene R and V7 m
dilute to 100.0 mL with die mobile phase.
Reference solution (a) Dissolve 10.0 mg of cholecalciferol CRS = mass of the preparation to be examined in the
without heating in 10.0 mL of toluene R and dilute to test solution, in milligrams;
100.0 mL with the mobile phase. = mass of cholecalciferol CRS in reference solution
(a), in milligrams;
Reference solution (b) Dilute 1.0 mL of cholecalciferolfor system
V = volume of the test solution (100 mL);
suitability CR S to 5.0 mL with the mobile phase. Heat in a
V = volume of reference solution (a) (100 mL);
water-bath at 90 °C under a reflux condenser for 45 min and
Sd = area (or height) of the peak due to cholecalciferol
cool.
in the chromatogram obtained with the test
Reference solution (c) Dissolve 0.10 g of cholecalciferol CRS
solution;
without heating in toluene R and dilute to 100.0 mL with the = area (or height) of the peak due to cholecalciferol
same solvent. in the chromatogram obtained with reference
Reference solution (d) Dilute 5.0 mL of reference solution (c) solution (a);
to 50.0 mL with the mobile phase. Keep the solution in iced = area (or height) of the peak due to pre-
water. cholecalciferol in the chromatogram obtained
Reference solution (e) Place 5.0 mL of reference solution (c) in with the test solution;
a volumetric flask, add about 10 mg of butylhydroxytoluene R f = conversion factor.
and displace air from the flask with nitrogen R. Heat in a
STORAGE
water-bath at 90 °C under a reflux condenser protected from
light and under nitrogen R for 45 min. Cool and dilute to ' In an airtight, well-filled container, protected from light.
50.0 mL with the mobile phase. The contents of an opened container are to be used as soon
as possible; any unused part is to be protected by an
Column'.
atmosphere of nitrogen.
— size. I = 0.25 m, 0 = 4.6 mm;
— stationary phase, silica gel for chromatography R (5 |im). LABELLING
Mobile phase pentanol R, hexane R (3:997 VIV). The label states:
— the number of International Units per gram;
Flow rate 2 mlVmin.
— the method of restoring the solution if partial
Detection Spectrophotometer at 254 nm. solidification occurs.
Injection The chosen volume of each solution (the same PhEur
volume for reference solution (a) and for the test solution);
automatic injection device or sample loop recommended.
Relative retention With reference to cholecalciferol: pre-
cholecalciferol = about 0.4; mzra-cholecaldferol = about 0.5.
2016 Colecalciferol 1-643
Reference solution (b) Dilute 1.0 mL of cholecalciferolfor system Sd = area (or height) of the peak due to cholecalciferol in
suitability CRS to 5.0 mL with the mobile phase. Heat in a the chromatogram obtained with the test solution;
water-bath at 90 °C under a reflux condenser for 45 min and S'd = area (or height) of the peak due to cholecalciferol in
cool. the chromatogram obtained with reference solution
Reference solution (c) Dissolve 0.10 g of cholecalciferol CRS, (a);
without heating, in toluene R and dilute to 100.0 mL with the Sp = area (or height) of the peak due to pre-
same solvent. cholecalciferol in the chromatogram obtained with
the test solution;
Reference solution (d) Dilute 5.0 mL of reference solution (c)
/ = conversion factor.
to 50.0 mL with die mobile phase. Keep the solution in iced
water. STORAGE
Reference solution (e) Place 5.0 mL of reference solution (c) in In an airtight, well-filled container, protected from light
a volumetric flask, add about 10 mg of butyJhydroxytoluene R The contents of an opened container are to be used as soon
and displace the air from the flask with nitrogen R. Heat in a as possible; any unused part is to be protected by an
water-bath at 90 °C under a reflux condenser, protected from atmosphere of nitrogen.
light and under nitrogen R, for 45 min. Cool and dilute to LABELLING
50.0 mL with the mobile phase. The label states the number of International Units per gram.
Column:
___________________________________________________________________________________________ PhEur
— size. I = 0.25 m, 0 = 4.6 mm;
— stationary phase, silica gelfor chromatography R (5 |im).
Mobile phase pentanol R, hexane R (3:997 VIV).
Flow rate 2 mL/min.
Detection Spectrophotometer at 254 nm. Colecalciferol Concentrate (Water- *****
Injection The chosen volume of each solution (the same dispersible Form) *****
volume for reference solution (a) and for the test solution); (Cholecalciferol Concentrate (Water-Dispersible Form),
automatic injection device or sample loop recommended. Ph Eur monograph 0598)
Relative retention With reference to cholecalciferol: pre-
cholecalciferol = about 0.4; iram-cholecalciferol = about 0.5. Action and use
System suitability: reference solution (b): Vitamin D analogue (Vitamin D3).
— resolution: minimum 1.0 between the peaks due to pre-
Ph E u r ___________________________________________________________________________________________
cholecalciferol and irons-cholecalciferol; if necessary,
adjust the proportions of the constituents and the flow DEFINITION
rate of the mobile phase to obtain this resolution; Solution of Cholecalciferol (0072) in a suitable vegetable fatty
— repeatability: maximum relative standard deviation of oil, authorised by the competent authority, to which suitable
1.0 per cent for the peak due to cholecalciferol after solubilisers have been added.
6 injections. Content
Calculate the conversion factor (f) using the following 90.0 per cent to 115.0 per cent of the cholecalciferol content
expression: stated on the label, which is not less than 100 000 IU/g.
It may contain suitable stabilisers such as antioxidants.
K -L
CHARACTERS
M
Appearance
K = area (or height) of the peak due to cholecalciferol in Slightly yellowish liquid of variable opalescence and viscosity.
the chromatogram obtained with reference solution Highly concentrated solutions may become cloudy at low
(d); temperatures or form a gel at room temperature.
L — area (or height) of the peak due to cholecalciferol in IDENTIFICATION
the chromatogram obtained with reference solution First identification A , C, D
(e); Second identification A , B, D
M = area (or height) of the peak due to pre-cholecalriferol
in the chromatogram obtained with reference A. Thin-layer chromatography (2.2.27). Prepare the solutions
solution (e). immediately before use.
Test solution Place 10.0 mL of the test solution prepared for
The value of /determined in duplicate on different days may
the assay in a suitable flask and evaporate to dryness under
be used during the entire procedure.
reduced pressure by swirling in a water-bath at 40 °C. Cool
Calculate the content of cholecalciferol in International Units under running water and restore atmospheric pressure with
per gram using the following expression: nitrogen R. Dissolve the residue immediately in 0.4 mL of
ethylene chloride R containing 10 g/L of squalane R and
2L x Y- x gg ,+ (/ x Sp) x 40000 x 1000 0.1 g/L of btctylhydroxytoluene R.
V' m S'D Reference solution (a) Dissolve 10 mg of cholecalciferol CRS in
ethylene chloride R containing 10 g/L of squalane R and
m = mass of the preparation to be examined in the test 0.1 g/L of butylhydroxytoluene R and dilute to 4 mL with the
solution, in milligrams; same solution.
m' = mass of cholecalciferol CRS in reference solution (a),
Reference solution (b) Dissolve 10 mg of ergocalciferd CRS in
in milligrams;
ethylene chloride R containing 10 g/L of squalane R and
V = volume of the test solution (25 mL);
V = volume of reference solution (a) (100 mL);
2016 Colecalciferol 1-645
0.1 g/L of butylhydroxytoluene R and dilute to 4 mL with the 50 mL of pentane R. After separation, transfer the aqueous-
same solution. alcoholic layer to a 3rd separating funnel and the pentane
Plate T L C silica gel G plate R. layer to the 1st separating funnel, washing the 2nd separating
funnel with 2 quantities, each of 10 m T , of pentane R and
Mobile phase A 0.1 g/L solution of butylhydroxytoluene R in a
adding the washings to the 1st separating funnel. Shake the
mixture of equal volumes of cydohexane R and peroxide-free
aqueous-alcoholic layer with 50 mL of pentane R and add the
ether R.
pentane layer to the 1st funnel. Wash the pentane layer with
Application 20 |iL. 2 quantities, each of 50 mL, of a freshly prepared 30 g/L
Development Immediately, protected from light, over a path of solution of potassium hydroxide R in ethanol
15 cm. (10 per cent V/V) R, shaking vigorously, and then wash with
Drying h i air. successive quantities, each of 50 m T , of water R until the
Detection Spray with sulfuric acid R. washings are neutral to phenolphthalein. Transfer the washed
pentane extract to a ground-glass-stoppered flask. Evaporate
Results The chromatogram obtained with the test solution
the contents of the flask to dryness under reduced pressure
shows immediately a bright yellow principal spot, which
by swirling in a water-bath at 40 °C. Cool under ru nning
rapidly becomes orange-brown, then gradually greenish-grey,
water and restore atmospheric pressure with nitrogen R.
rem a in in g so for 10 m in . This spot is sim ilar in position,
Dissolve the residue im m ediately in 5.0 mL of toluene R and
colour and size to the principal spot in the chromatogram
add 20.0 mL of the mobile phase to obtain a solution
obtained with reference solution (a). The chromatogram
containing about 4000 IU/mL.
obtained with reference solution (b) shows immediately at
the same level an orange principal spot, which gradually Reference solution (a) Dissolve 10.0 mg of cholecalciferol CRS ,
becomes reddish-brown and remains so for 10 min. without heating, in 10.0 mL of toluene R and dilute to
100.0 mL with the mobile phase.
B. Ultraviolet and visible absorption spectrophotometry
(2.2.25). Reference solution (b) Dilute 1.0 mL of cholecalciferolfor system
suitability CRS to 5.0 mL with the mobile phase. Heat in a
Test solution Place 5.0 mL of the test solution prepared for
water-bath at 90 °C under a reflux condenser for 45 min and
the assay in a suitable flask and evaporate to dryness under
cool.
reduced pressure by swirling in a water-bath at 40 °C. Cool
under ru n n in g water and restore atmospheric pressure with Reference solution (c) Dissolve 0.10 g of cholecalciferol CRS,
nitrogen R. Dissolve the residue im m ediately in 50.0 mL of without heating, in toluene R and dilute to 100.0 mL with the
cycbhexane R. same solvent.
Spectral range 250-300 nm. Reference solution (d) Dilute 5.0 mL of reference solution (c)
to 50.0 mL with the mobile phase. Keep the solution in iced
Absorption maximum At 265 nm.
water.
C. Examine the chromatograms obtained in the assay.
Reference solution (e) Place 5.0 mL of reference solution (c) in
Results The principal peak in the chromatogram obtained
a volumetric flask, add about 10 mg of butylhydroxytoluene R
with the test solution is similar in retention time to the and displace the air from the flask with nitrogen R. Heat in a
principal peak in the chromatogram obtained with reference water-bath at 90 °C under a reflux condenser, protected from
solution (a). light and under nitrogen R, for 45 min. Cool and dilute to
D. Mix about 1 g with 10 mL of water R previously wanned 50.0 mL with the mobile phase.
to 50 °C, and cool to 20 °C. Immediately after cooling, a Column:
uniform, slightly opalescent and slightly yellow dispersion is — sizer. I = 0.25 m, 0 = 4.6 mm;
obtained. — stationary phase: silica gelfor chromatography R (5 fun).
TESTS Mobile phase pentanol R, hexane R (3:997 VIV).
Related substances Flow rate 2 mL/min.
The thresholds indicated under Related substances
Detection Spectrophotometer at 254 nm.
(Table 2034.-1) in the general monograph Substancesfor
pharmaceutical use (2034) do not apply. Injection The chosen volume of each solution (the same
volume for reference solution (a) and for the test solution);
ASSAY automatic injection device or sample loop recommended.
Cany out the assay as rapidly as possible, avoiding exposure to
Relative retention With reference to cholecalciferol: pre-
actinic light and air.
cholecalciferol = about 0.4; mzw-cholecalciferol = about 0.5.
Liquid chromatography (2.2.29). System suitability: reference solution (b):
Test solution Introduce into a saponification flask a quantity of — resolution: minimum 1.0 between the peaks due to pre-
the preparation to be examined, weighed with an accuracy of cholecalciferol and mim-cholecalciferol; if necessary,
0.1 per cent, equivalent to about 100 000 IU. Add 5 mL of adjust the proportions of the constituents and the flow
water R, 20 mL of anhydrous ethanol R, 1 mL of sodium rate of the mobile phase to obtain this resolution;
ascorbate solution R and 3 mL of a freshly prepared — repeatability: maximum relative standard deviation of
50 per cent m/m solution of potassium hydroxide R. Heat in a 1.0 per cent for the peak due to cholecalciferol after
water-bath under a reflux condenser for 30 min. Cool rapidly 6 injections.
under running water. Transfer the liquid to a separating Calculate the conversion factor (f) using the following
funnel with the aid of 2 quantities, each of 15 ml., of expression:
water R, 1 quantity of 10 mL of ethanol (96 per cent) R and
2 quantities, each of 50 mT, of pentane R. Shake vigorously
K -L
for 30 s. Allow to stand until the 2 layers are clear. Transfer
the aqueous-alcoholic layer to a 2nd separating funnel and M
shake with a mixture of 10 mL of ethanol (96 per cent) R and
1-646 Colestipol Hydrochloride 2016
K — area (or height) of the peak due to cholecalciferol Practically insoluble in ethanol (96%) and in dichloromethaner,
in the chromatogram obtained with reference swells but does not dissolve in water and dilute aqueous
solution (d); solutions of acids and alkalis.
L = area (or height) of the peak due to cholecalciferol IDENTIFICATION
in the chromatogram obtained with reference
Carry out the method for gas chromatography,
solution (e);
Appendix m 5, using a suitable gas chromatograph fitted
M = area (or height) of the peak due to pre-
with a pyrolysis unit Operate the unit in accordance with the
cholecalciferol in the chromatogram obtained with
manufacturer’s instructions to obtain a pyrogram for colestipol
reference solution (e).
hydrochloride BPCRS that is similar to that supplied with the
The value of /determined in duplicate on different days may reference material.
be used during the entire procedure. (1) To prepare the sample, mix 1 part of n -eicosane and 4
Calculate the content of cholecalciferol in International Units parts of the substance being examined and grind the mixture
per gram vising the following expression: in a mortar with chloroform until the substance being
examined is un iform ly coated with the n-eicosane.
V w SD + (/ x Sp) (2) Prepare the standard in the same manner but adding 4
177 x x 40 000 x 1000
V' x ~
m parts of colestipol hydrochloride BPCRS in place of the
substance being examined.
mass of the preparation to be exam ined in the C H R O M A T O G R A P H IC C O N D IT IO N S
test solution, in milligrams;
(a) Use a a glass column (1.8 m x 3 mm) packed with acid-
mass of cholecalciferol CRS in reference solution
washed, sUattised diatomaceous support (80 to 100 mesh)
(a), in milligrams;
V volume of the test solution (25 mL); (Chromosorb W is suitable) coated with 0.25% w/w of
potassium hydroxide and 5% w/w of polyethylene glycol 20,000
V volume of reference solution (a) (100 mL);
(Carbowax 20M is suitable).
SD area (or height) of the peak due to cholecalciferol
in the chromatogram obtained with the test (b) Use helium as the carrier gas at 60 mL per minute.
solution; (c) Use isothermal conditions maintained at 85°.
S 'D = area (or height) of the peak due to cholecalciferol (d) Use a pyrolysis unit capable of attaining a temperature of
in the chromatogram obtained with reference about 1000° when fined with a platinum ribbon probe.
solution (a); (e) Use a detector at a temperature of 270°.
area (or height) of the peak due to pre-
cholecalciferol in the chromatogram obtained (f) Load the sample and the standard separately into the
with the test solution; pyrolysis unit.
f conversion factor. C O N F IR M A T IO N
and 0.25 mL of sulfuric acid and heat cautiously until white (m g) of the substance being examined to a ground-glass-
fames are no longer evolved. Ignite, preferably in a muffle stoppered flask, add 100 mL of freshly prepared solution A
furnace, at 500° to 600°, until the carbon is completely and shake vigorously for 90 minutes with the flask positioned
removed and cool. Add 4 mL of 6m hydrochloric add, cover, horizontally. Remove the flask from the shaker and allow the
heat on a water bath for 15 minutes, uncover and slowly contents to settle for 5 minutes. Adjust the pH of a 20 mL
evaporate to dryness. Dissolve the residue using two 5 mL aliquot of the supernatant liquid to 10.5 by the drop wise
quantities of 2m hydrochloric acid. Add 0.1 mL of addition of 1 m sodium hydroxide and titrate potentiometrically
phenolphthalein solution and 13.5m ammonia drop wise until a with 0. 1m hydrochloric acid VS to the second inflection point
pink colour is produced. Cool, add glacial acetic add until the of the pH curve. Determine the volume of titrant added
solution is decolourised and add a further 0.5 mT- Filter if between the inflection points. Carry out a blank titration on
necessary and dilute the solution to 20 mL with water. 20 mL of freshly prepared solution A. The difference
12 mL of die resulting solution complies with limit test A for between the titrations represents the amount of hydrochloric
heavy metals, Appendix VII (20 ppm). Prepare the standard acid required (F mL). Calculate the cholate binding capacity
using a mixture of 2 mL of the test solution obtained above of the substance being examined in milliequivalents from the
and 10 mL of lead standard solution (2 ppm Pb). expression 0.5V/m.
Loss on drying Colestipol exchange capacity lim it
When dried at 75° at a pressure of not more than 2 kPa for Not less than 9.0 mEq per g and not more than 11.0 mEq
16 hours, loses not more than 1.0 % of its weight per g determined in the following manner. Transfer not less
Sulfated ash than 2 g and 100 mL of 1m sodium hydroxide to a stoppered
Not more than 0.3%, Appendix IX A. flask and shake for 4 hours. Filter the suspension through a
coarse-porosity sintered-glass funnel and wash the resin with
Chloride content 500 mL of water. Transfer the resin to a 1000 mL beaker,
Not less than 6.5% and not more than 9.0% calculated with add 200 mL of water and allow to stand for 10 minutes.
reference to the dried substance. Bum 20 mg by the method Filter the suspension, check the pH of the filtrate and repeat
for oxygen-flask combustion, Appendix VHI C, using 10 mL of the washing procedure with 200 mL quantities of water until
0.05m sodium hydroxide as the absorbing liquid. When the
the pH of the filtrate is less than 8 [5 litres may be required].
process is complete shake the flask vigorously, allow to stand Dry the resin and funnel at 60° at a pressure of 2 kPa for at
with frequent shaking for about 40 minutes or until no least 16 hours, breaking up any aggregates with a spatula and
cloudiness is observed; add 20 mL of ethanol (96%) and store in a dessicator.
0.2 mL of nitric acid. Titrate the resulting solution with
0.05m silver nitrate FS, determining the end point
Place 1 g (w g) of the free base resin prepared above and add
100 mL of 0.2m hydrochloric acid FS in a stoppered flask and
potentiometrically using a silver-silver chloride electrode and
a glass reference electrode (FmL). Repeat the procedure shake for not less than 2.5 hours. Filter a portion of the
without the substance being examined adding 10 mL of suspension through glass wool. Titrate 8 mL of the filtrate
0.0075m sodium chloride to the solution in the flask (Vx mL).
with 0.2m sodium hydroxide FS, determining the end point
Add 10 mL of 0.0075m sodium chloride to a flask containing a potentiometrically (a mL). Carry out a blank titration on
5 mL of the 0.2m hydrochloric acid FS used to equilibrate the
mixture of 10 mL of water and 20 mL of ethanol (96%), add
free-base resin diluted with 5 mL of water (b mL). Calculate
0.2 mL of nitric acid and titrate with 0.05m silver nitrate FS,
determining the end point potentiometrically using a silver- the exchange capacity in milliequivalents per g from the
silver chloride electrode and a glass reference electrode expression:
(F 2 mL). Determine the volume of 0.05m silver nitrate FS (20/w)(b/5—a/8)
required by the substance being examined using the following
expression: STORAGE
V-(Vr V2)
Colestipol Hydrochloride should be kept in an airtight
container.
Each mL of 0.05m silver nitrate FS is equivalent to 1.773 mg
of Cl.
W ater absorption capacity
Each g of Colestipol Hydrochloride absorbs not less than Colestyramine *****
3.3 g and not more than 5.3 g of water when determined in
+*
the following manner. Transfer 5 g to a dry, plastic container (Ph. Eur. monograph 1775) *
and add 80 g of water. Cover the container and allow the 11041-12-6
resulting suspension to equilibrate for 72 hours. Filter the
resulting slurry through a medium-porosity fritted-glass Action and use
funnel (KLMAX 60 mL-40M is suitable) at a pressure of Lipid-regulating drug.
2kPa; collect the filtrate in a tared, plastic container, Preparation
disconnecting the vacuum 2 minutes after collection of the Colestyramine Oral Powder
last portion of the filtrate. Immediately weigh the container
and the filtrate and determine the weight, in g, of the filtrate. PhEur_____________________ :____________________________________
Calculate the weight of water absorbed per g from the
DEFINITION
difference between the weight of the filtrate and the original
Strongly basic anion-exchange resin in chloride form,
weight of water used in the test
consisting of styrene-divinylbenzene copolymer with
Cholate binding capacity quaternary ammonium groups.
Prepare a solution containing 1.0 % w/v of sodium cholate and
Nominal exchange capacity 1.8 g to 2.2 g of sodium
0.9% w/v of sodium chloride and adjust to pH 6.4 by the drop
glycocholate per gram (dried substance).
wise addition of hydrochloric acid (solution A). Transfer 1 g
1-648 Colestyramine 2016
CHARACTERS Column:
Appearance — sizer. I = 0.30 m, 0 = 3.9 mm,
White or almost white, fine powder, hygroscopic. — stationary phase: octadecylsQyl silica gelfor chromatography R
Solubility (10 pm) with a specific surface area of 330 m2/g and a
Insoluble in water, in methylene chloride and in ethanol pore size of 12.5 nm.
(96 per cent). Mobile phase acetomtrUe R, water R (50:50 VIV).
Flow rate 2.0 mL/min.
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24). Detection Spectrophotometer at 254 nm.
Comparison colestyramine CRS. Injection 20 |iL of test solution and reference solutions (a)
and (d).
B. Chloride (see Tests).
System suitability Reference solution (d):
TESTS — resolution: minimum 1.5 between the peaks due to
pH (2.2.5) impurity A and toluene.
4.0 to 6.0. Limir.
Suspend 0.100 g in 10 mL of water R and allow to stand for — impurity A: not more than the area of the principal peak
10 min. in the chromatogram obtained with reference solution (a)
Dialysable quaternary amines (1 ppm).
Maximum 500 ppm, expressed as benzyltrimethylammonium Chloride
chloride. 13.0 per cent to 17.0 per cent (dried substance).
Test solution Place a 25 cm piece of cellulose dialysis tubing To 0.2 g add 100 mL of water R and 50 mg of potassium
having a molecular weight cut-off of 12 000-14 000 and an nitrate R. Add, with stirring, 2 mL of nitric add R and titrate
inflated diameter of 3-6 cm (flat width of 5-9 cm) in water R with 0.1 M silver nitrate, determining the end-point
to hydrate until pliable, appropriately sealing one end. potentiometrically (2.2.20).
Introduce 2.0 g of the substance to be examined into the 1 mL of 0.1 M silver nitrate is equivalent to 3.55 mg of CL
tube and add 10 mL of water R. Seal the tube and
completely immerse it in 100 mL of water R in a suitable Heavy metals (2.48)
vessel and stir the liquid for 16 h to effect dialysis. Use the Maximum 20 ppm.
dialysate as test solution. 1.0 g complies with test F. Prepare the reference solution
Reference solution Prepare the reference solution in a similar
using 2 mL of lead standard solution (10 ppm Pb) R.
manner but using 10 mL of a freshly prepared 0.1 g/L Loss on drying (2.2.32)
solution of benzyltrimethylammonium chloride R instead of the Maximum 12 per cent, determined on 1.000 g by drying in
substance to be examined. an oven at 70 °C over diphosphorus pentoxide R at a pressure
Transfer 5.0 mL of the test solution to a separating funnel not exceeding 7 kPa for 16 h.
and add 5 ml. of a 3.8 g/L solution of disodium tetraborate R, Sulfated ash (2.414)
1 mL of a solution containing 1.5 g/L of bromothymol blue R Maximum 0.1 per cent, determined on 1.0 g.
and 4.05 g/L of sodium carbonate R and 10 mL of ASSAY
chloroform R. Shake the mixture vigourously for 1 min, allow
Exchange capacity
the phases to separate and transfer the clear organic layer to
Liquid chromatography (2.2.29).
a 25 mL volumetric flask. Repeat the extraction with a
Solution A Dissolve 1.500 g of sodium glycocholate R in a
further 10 mL of chloroform R, combine the organic layers
and dilute to 25 mL with chloroform R. Measure die solution containing 4 g/L of potassium dihydrogen phosphate R
absorbance (2.2.25) of the solution at the absorption and 12 g/L of dtpotassiitm hydrogen phosphate R and dilute to
maximum at 420 nm, using as compensation liquid a 100.0 mL with die same solution.
solution prepared in the same manner but using 5.0 mL of Test solution Add 20.0 mL of solution A to a quantity of die
water R instead of the test solution. substance to be examined equivalent to about 0.100 g of the
Repeat the operation using 5.0 mL of the reference solution. dried substance. Shake mechanically for 2 h and centrifuge
for 15 min. Dilute 5.0 mL of the supernatant to 50.0 mL
The absorbance obtained with the test solution is not greater
with water R.
than that obtained with the reference solution.
Reference solution (a) Dilute 4.0 mL of solution A to
Im purity A 100.0 mL with water R.
Liquid chromatography (2.2.29).
Reference solution (b) Dissolve 60 mg of sodium glycocholate R
Test solution Shake 5.0 g with 10 mL of acetone R for 30 min.
and 30 mg of sodium taurodeoxychdate R in water R and
Centrifuge and use the supernatant. dilute to 100 mL with the same solvent Dilute 1 mL of the
Reference solution (a) Dissolve 5 mg of styrene R in acetone R solution to 10 mL with water R.
and dilute to 100.0 mL with the same solvent. Dilute 1.0 mL Column:
of the solution to 100.0 mL with acetone R. — size: I = 0.25 m, 0 = 4.6 mm,
Reference solution (b) Dissolve 0.35 mL of styrene R in — stationary phase: octadecylsüyl silica gelfor chromatography R
acetone R and dilute to 100.0 mL with the same solvent. (5 pm).
Dilute 1.0 mL of the solution to 100.0 mL with acetone R. Mobile phase Mix 35 volumes of acetonitrüe R and 65 volumes
Reference solution (c) Dissolve 0.35 mL of toluene R in of a 10.9 g/L solution of potassium dihydrogen phosphate R
acetone R and dilute to 100.0 mL with the same solvent. adjusted to pH 3.0 with phosphoric acid R.
Reference solution (d) Mix 1.0 mL of reference solution (b) Flow rate 1.5 mL/min.
and 1.0 mL of reference solution (c) with acetone R and Detection Spectrophotometer at 214 nm.
dilute to 100.0 mL with the same solvent.
2016 Colistimethate Sodium 1-649
Composition Sulfate
Liquid chromatography (2.2.29): use the normalisation 16.0 per cent to 18.0 per cent (dried substance).
procedure. Dissolve 0.250 g in 100 mL of water R and adjust to pH 11
Test solution Dissolve 25.0 mg of the substance to be with concentrated ammonia R. Add 10.0 m l . of 0.1 M barium
examined in 40 mL of water R and dilute to 50.0 ml. with chloride and about 0.5 mg of phthalein purple R. Titrate with
acetomtrUe R l. 0.1 M sodium edetaxe, adding 50 mL of ethanol (96 per cent) R
Reference solution (a) Dissolve 25.0 mg of colistin sulfate CRS when the colour of the solution begins to change and
in 40 mL of water R and dilute to 50.0 mL with continuing the titration until the violet-blue colour
acetonitrUe R l. disappears.
Dilute 1.0 mL of reference solution (a)
Reference solution (b) 1 mL of 0.1 M barium chloride is equivalent to 9.606 mg
to 100.0 mL with a mixture of 20 volumes of acetonitrUe R l of S04.
and 80 volumes of water R. Loss on drying (2.2.52)
Column: M axim um 3.5 per cent, determined on 1.000 g by drying at
— size: I = 0.15 m, 0 = 4.6 mm; 60 °C over diphosphorus pentoxide R at a pressure not
— stationary phase: end-capped octadecylsUyl sUica gel for exceed in g 0.67 kPa for 3 h.
chromatography R (3.5 jun); Sulfated ash (2.4.14)
— temperature: 30 °C. Maximum 1.0 per cent, determined on 1.0 g.
Mobile phase Mix 22 volumes of acetomtrUe R l and ASSAY
78 volumes of a solution prepared as follows: dissolve 4.46 g Carry out the microbiological assay of antibiotics (2.7.2).
of anhydrous sodium sulfate R in 900 mL of water R, adjust to
STORAGE
pH 2.4 with dilute phosphoric add R and dilute to 1000 ml.
with water R. In an airtight container, protected from light
__________________________________________________________PhEur
Flow rate 1.0 mL/min.
Detection Spectrophotometer at 215 nm.
Injection 20 jiL.
Copovidone ★ ★
Run time 1.5 times the retention time of polymyxin El. ★ ★
★ ★
Identification ofpeaks Use the chromatogram supplied with (Ph Eur monograph 0891) ***
colistin sulfate CRS to identify the peaks due to polymyxins
El, E2, E3, E l-I and E1-7MOA.
Relative retention With reference to polymyxin E l (retention
time = about 16 mm): polymyxin E2 = about 0.45;
polymyxin E3 = about 0.5; polymyxin El-I = about 0.8;
polymyxin E1-7MOA = about 1.1.
System suitabUity. reference solution (a):
— resolution: minimum 8.0 between the peaks due to (QHçNO)*, (C4H60 2)m Mt(lll.l) n + (86 . 1)„ 25086-89-9
polymyxin E2 and polymyxin El; minimum 6.0 between
the peaks due to polymyxin E2 and polymyxin El-I; Action and use
minimum 2.5 between the peaks due to polymyxin E l-I Excipient in pharmaceutical products.
and polymyxin E l; minimum 1.5 between the peaks due
PhEur.
to polymyxin E l and polymyxin E1-7MOA.
Limity. DEFINITION
— polymyxin E3: maximum 10.0 per cent (dried substance); Copovidone is a copolymer of 1-ethenylpyrrolidin-2 -one and
— polymyxin El-I: maximum 10.0 per cent (dried ethenyl acetate in the mass proportion 3:2.
substance); Content
— polymyxin E1-7MOA: maximum 10.0 per cent (dried — nitrogen (N; A r 14.01): 7.0 per cent to 8.0 per cent
substance); (dried substance),
— sum ofpolymyxins E l, E2, E3, E l-I and E1-7MOA: — ethenyl acetate CjHeO^ 86.10): 35.3 per cent to
minimum 77.0 per cent (dried substance); 42.0 per cent (dried substance).
— disregard limit. the area of the peak due to polymyxin E l
K-value 90.0 per cent to 110.0 per cent of the value stated
in the chromatogram obtained with reference solution (b) on the label.
( 1.0 per cent).
CHARACTERS
Related substances
Liquid chromatography (2.2.29) as described in the test for Aspect,white or yellowish-white hygroscopic powder or flakes.
composition with the following modifications. Use the Solubility
normalisation procedure. Freely soluble in water, in ethanol (96 per cent) and in
Injection Test solution and reference solution (b). methylene chloride.
Limits: IDENTIFICATION
— any impurity, maximum 4.0 per cent; First identification: A.
— total: maximum 23.0 per cent; Second identification B, C
— disregard limit, the area of the peak
due to polymyxin El A. Infrared absorption spectrophotometry (2.2.24).
in the chromatogram obtained with reference solution (b);
Comparison Ph. Eur. reference spectrum of copovidone.
disregard the peaks due to polymyxins E l, E2, E3, E l-I
and E1-7MOA. B. To 1 mL of solution S (see Tests) add 5 mL of water R
and 0.2 mL of 0.05 M iodine. A red colour appears.
1-652 Copovidone 2016
C. Dissolve 0.7 g of hydroxylamine hydrochloride R in 10 mL Ai = absorbance of the reference solution before the
of methanol R, add 20 mL of a 40 g/L solution of sodium addition of aldehyde dehydrogenase;
hydroxide R and filter if necessary. To 5 mL of the solution ■
A-t2 = absorbance of the reference solution after the
add 0.1 g of the substance to be examined and boil for addition of aldehyde dehydrogenase;
2 min. Transfer 50 pL to a filter paper and add 0.1 mL of a Abi = absorbance of the blank before the addition of
mixture of equal volumes offerric chloride solution R l and aldehyde dehydrogenase;
hydrochloric acid R. A violet colour appears. A/,2 = absorbance of the blank after the addition of
aldehyde dehydrogenase;
TESTS
m = mass of povidone, in grams, calculated with
Solution S
-reference to the dried substance;
Dissolve 10.0 g in water R and dilute to 100.0 mL with the
C = concentration (mg/ml), of acetaldehyde in the
same solvent. Add the substance to be examined to the
reference solution, calculated from the weight of
water R in small portions with constant stirring.
the acetaldehyde ammonia trimer trihydrate with
Appearance o f solution the factor 0.72.
Solution S is not more opalescent than reference
suspension HI (2.2.1) and not more intensely coloured than Peroxides
reference solution B5, R 5 or BY5 (2.2.2, Method II). Maximum 400 ppm, expressed as H 20 2.
Dilute 10 mL of solution S to 25 mL with water R.
Viscosity, expressed as ft-vahie
Add 2 mL of titanium trichloride-sulfuric acid reagent R and
Dilute 5.0 mL of solution S to 50.0 mL with water R. Allow
allow to stand for 30 min. The absorbance (2.2.25) of the
to stand for 1 h and determine the viscosity (2.2.9) of the
solution, measured at 405 nm using a mixture of 25 mL of a
solution at 25 ± 0.1 °C, using a size n° 1 viscometer with a
40 g/L solution of the substance to be examined and 2 mL of
minimum flow time of 100 s. Calculate the iC-value using the
a 13 per cent V/V solution of sulfuric acid R as the
following expression:
compensation liquid, is not greater than 0.35.
Hydrazine
1.51ogio V ~ 1 , \/300c l°gio r) + (c + l-5c l°gi° r?)2 Thin-layer chromatography (2.2.27). Usefreshly prepared
solutions.
0.15 + 0.003c + 0.15c + 0.003c2
Test solution To 25 mL of solution S add 0.5 mL of a 50 g/L
c = percentage concentration (g/100 mL) of the solution of salicylaldehyde R in methanol R, mix and heat in a
substance to be examined, calculated with reference water-bath at 60 °C for 15 min. Allow to cool, add 2.0 mL
to the dried substance; of xylene R, shake for 2 min and centrifuge. Use the clear
rj = viscosity of the solution relative to that of water. supernatant layer.
Reference solution Dissolve 9 mg of salicylaldehyde azine R in
Aldehydes
xylene R and dilute to 100 mL with the same solvent. Dilute
Maximum 500 ppm, expressed as acetaldehyde.
1 mL of the solution to 10 mL with xylene R.
Test solution Dissolve 1.0 g of the substance to be examined
Hate TLC sUamsed silica gel plate R.
in phosphate buffer solution p H 9.0 R and dilute to 100.0 mL
with the same solvent. Stopper the flask and heat at 60 °C Mobile phase water R, methanol R (20:80 VtV).
for 1 h. Allow to cool. Application 10 jiL.
Reference solution Dissolve 0.140 g of acetaldehyde ammonia Development Over 3/4 of the plate.
trimer trihydrate R in water R and dilute to 200.0 mL with the Drying In air.
same solvent. Dilute 1.0 mL of the solution to 100.0 mL Detection Examine in ultraviolet light at 365 nm.
with phosphate buffer solution pH 9.0 R. Limit:
Into 3 identical spectrophotometric cells with a path length — hydrazine: any spot due to salicylaldehyde azine is not
of 1 cm, introduce separately 0.5 mL of the test solution, more intense than the spot in the chromatogram obtained
0.5 mL of the reference solution and 0.5 mL of water R with the reference solution (1 ppm).
(blank). To each cell add 2.5 mL of phosphate buffer solution
pH 9.0 R and 0.2 mL of nicotmamide-adenme dinucleotide
Monomers
solution R. Mix and stopper tightly. Allow to stand at
Maximum 0.1 per cent.
22 ± 2 °C for 2-3 min and measure the absorbance (2.2.25) Dissolve 10.0 g in 30 mT. of methanol R and add slowly
of each solution at 340 nm, using water R as the 20.0 mL of iodine bromide solution R. Allow to stand for
compensation liquid. To each cell, add 0.05 mL of aldehyde 30 min protected from light with repeated shaking.
dehydrogenase solution R, mix and stopper tightly. Allow to Add 10 mL of a 100 g/L solution of potassium iodide R and
stand at 22 ± 2 °C for 5 min. Measure the absorbance of titrate with 0.1 M sodium tMosulfate until a yellow colour is
each solution at 340 nm using water R as compensation obtained. Continue titration dropwise until the solution
liquid. Determine the content of aldehydes using the becomes colourless. Carry out a blank titration. Not more
following expression: than 1.8 mL of 0.1 M sodium thiosulfate is used.
Im purity A
(At 2 — A t\) — (Ab2 — A\,x)100 000 x C Liquid chromatography (2.2.29).
(A, 2 —At \) — (Ab2 —A n ) m Test solution Dissolve 0.100 g of the substance to be
examined in water R and dilute to 50.0 mL with the same
A ti = absorbance of the test solution before the addition solvent.
of aldehyde dehydrogenase; Reference solution Dissolve 0.100 g of 2-pyrrolidone R
A [2 = absorbance of the test solution after the addition (impurity A) in water R and dilute to 100 mL with the same
of aldehyde dehydrogenase; solvent. Dilute 1.0 ml. to 100.0 mL with water R.
2016 Copper Sulfate 1-653
Iron Solubility
Maximum 150 ppm. Freely soluble in water, soluble in methanol, practically
Atomic absorption spectrometry (2.2.25, Method I). insoluble in ethanol (96 per cent).
Test solution Dissolve 0.32 g in 10 mL of water R, add IDENTIFICATION
2.5 mL of lead-free nitric acid R and dilute to 25.0 mL with A. Add several drops of dilute ammonia R2 to 1 mL of
water R. solution S (see Tests). A blue precipitate is formed.
Reference solutions Prepare the reference solutions using iron On further addition of dilute ammonia R2 the precipitate
standard solution (20 ppm Fe) R, adding 2.5 mL of lead-free dissolves and a dark blue colour is produced.
nitric add R and diluting to 25.0 mL with water R. B. Loss on drying (see Tests).
Source Iron hollow-cathode lamp. C. Dilute 1 mL of solution S to 5 mL with water R.
Wavelength 248.3 nm. The solution gives reaction (a) of sulfates (2.3.1).
Atomisation device Air-acetylene flame. TESTS
Copper may form explosive acetyUdes with acetylene. Therefore, Solution S
clean the burner thoroughly before any residues become dry. Dissolve 5 g in water R and dilute to 100 mL with the same
Lead solvent.
Maximum 80 ppm. Appearance of solution
Atomic absorption spectrometry (2.2.25, Method I). Solution S is clear (2.2.1).
Test solution Dissolve 1.6 g in 10 mL of water R, add 2.5 mL Chlorides (2.4.4)
of lead-free nitric add R and dilute to 25.0 mL with water R. M axim um 100 ppm.
Reference solutions Prepare the reference solutions using lead Dilute 10 mL of solution S to 15 m l . with water R.
standard solution (100 ppm Pb) R, adding 2.5 mL of lead-free Iron
nitric add R and diluting to 25.0 ml. with water R. Maximum 100 ppm.
Source Lead hollow-cathode lamp. Atomic absorption spectrometry (2.2.23, Method I).
Wavelength 217.0 nm. Test solution Dissolve 0.5 g in 10 mL of water R, add 2.5 mL
Atomisation device Air-acetylene flame. of lead-free nitric add R and dilute to 25.0 mL with water R.
Copper may form explosive acetyUdes with acetylene. Therefore, Reference solutions Prepare the reference solutions using iron
dean the burner thoroughly before any residues become dry. standard solution (20 ppm Fe) R, adding 2.5 mL of lead-free
nitric add R and diluting to 25.0 m l . with water R.
Loss on drying (2.2.52)
Maximum 1.0 per cent, determined on 0.500 g by drying in Source Iron hollow-cathode lamp.
an oven at 250 ± 10 °C. Wavelength 248.3 nm.
A. Infrared absorption spectrophotometry (2.2.24). Mobile phase Add a mixture of 1.2 volumes of water R and
8 volumes of methanol R to a mixture of 15 volumes of
Comparison cortisone acetate CRS.
ether R and 77 volumes of methylene chloride R.
If the spectra obtained in the solid state show differences,
Application 5 |iL.
record new spectra using 50 g/L solutions in methylene
chloride R in a 0.2 mm celL Development Over a path of 15 cm.
B. Thin-layer chromatography (2.2.27). Drying In air.
Solvent mixture methanol R, methylene chloride R (1:9 VIV). Detection A Examine in ultraviolet light at 254 nm.
Test solution Dissolve 10 mg of the substance to be examined Results A The principal spot in each of the chromatograms
in the solvent mixture and dilute to 10 mL with the solvent obtained with the test solutions is similar in position and size
mixture. to the principal spot in the chromatogram obtained with the
corresponding reference solution.
Reference solution (a) Dissolve 20 mg of cortisone acetate CRS
in the solvent mixture and dilute to 20 mL with the solvent Detection B Spray with alcoholic solution of sulfuric acid R and
mixture. heat at 120 °C for 10 min or until the spots appear. Allow to
cool. Examine in daylight and in ultraviolet light at 365 nm.
Reference solution (b) Dissolve 10 mg of hydrocortisone
acetate R in reference solution (a) and dilute to 10 mL with Results B The principal spot in each of the chromatograms
reference solution (a). obtained with the test solutions is similar in position, colour
in daylight, fluorescence in ultraviolet light at 365 nm and
Plate T LC silica gel F 254 plate R.
size to the principal spot in the chromatogram obtained with
Mobile phase Add a mixture of 1.2 volumes of water R and the corresponding reference solution. The principal spots in
8 volumes of methanol R to a mixture of 15 volumes of the chromatograms obtained with test solution (b) and
ether R and 77 volumes of methylene chloride R. reference solution (b) have an RP value distinctly lower than
Application 5 (iL. that of the principal spots in the chromatograms obtained
Development Over a path of 15 cm. with test solution (a) and reference solution (a).
Drying In. air. D. Add about 2 mg to 2 mL of sulfuric acid R and shake to
Detection A Examine in ultraviolet light at 254 nm. dissolve. Within 5 min, a faint yellow colour develops.
Add this solution to 10 mL of water R and mix. The colour
Results A The principal spot in the chromatogram obtained
is discharged and a clear solution remains.
with the test solution is similar in position and size to the
principal spot in the chromatogram obtained with reference E. About 10 mg gives the reaction of acetyl (2.3.1).
solution (a).
1-656 Cottonseed Oil 2016
TESTS
Specific optical rotation (2.2.7)
+ 211 to + 220 (dried substance).
Dissolve 0.250 g in dioxan R and dilute to 25.0 mL with the
same solvent.
Related substances
Liquid chromatography (2.2.29). Prepare the solutions A. ll|3,17-dihydroxy-3,20-dioxopregn-4-en-21-yl acetate
immediately before use. (hydrocortisone acetate).
Test solution Dissolve 25.0 mg of the substance to be _____________________________________________ PhEur
examined in acetomtrile R and dilute to 10.0 mL with the
same solvent.
Reference solution (a). Dissolve 2 mg of cortisone acetate CRS
and 2 mg of hydrocortisone acetate CRS (impurity A) in
acetomtrile R and dilute to 100.0 mL with the same solvent
Hydrogenated Cottonseed Oil *****
Reference solution (b) Dilute 1.0 mL of the test solution to (Ph. Eur. monograph 1305) *
100.0 mL with acetonitrUe R. PhEur_____________________________________________
Column:
— sizer. I = 0.25 m, 0 = 4.6 mm; DEFINITION
— stationary phase: octadecylsUyl sUica gel for chromatography R Product obtained by refining and hydrogenation of oil
(5 pm). obtained from seeds of cultivated plants of various varieties of
Gossypium hirsutum L. or of other species of Gossypium.
MobUe phase In a 1000 mL volumetric flask mix 400 mL of
The product consists mainly of triglycerides of palmitic and
acetomtrUe R with 550 mL of water R and allow to
stearic adds.
equilibrate; dilute to 1000 mL with water R and mix again.
Flow rate 1 mlVmin. CHARACTERS
Appearance
Detection Spectrophotometer at 254 nm.
White or almost white mass or powder which melts to a
Equilibration With the mobile phase for about 30 min.
clear, pale yellow liquid when heated.
Injection 20 jiL; inject acetomtrUe R as a blank.
Solubility
Run time Twice the retention time of cortisone acetate. Practically insoluble in water, freely soluble in methylene
Retention time Impurity A = about 10 min; cortisone chloride and in toluene, very slightly soluble in ethanol
acetate = about 12 min. (96 per cent).
System suitability: reference solution (a): IDENTIFICATION
— resolution: m in im u m 4.2 between the peaks due to A. Mdting point (see Tests).
impurity A and cortisone acetate; if necessary, adjust the
concentration of acetomtrile in the mobile phase. B. Composition of fatty adds (see Tests).
Limits: TESTS
— impurity A: not more than 0.5 times the area of the Melting point (2.2.14)
principal peak in the chromatogram obtained with 57 °C to 70 °C.
reference solution (b) (0.5 per cent); A d d value (2.5.1)
— total: not more than 1.5 times the area of the principal Maximum 0.5.
peak in the chromatogram obtained with reference Dissolve 10.0 g in 50 mT. of a hot mixture of equal volumes
solution (b) (1.5 per cent); of ethanol (96 per cent) R and toluene R , previously neutralised
— disregard limit: 0.05 times the area of the principal peak in with 0.1 M potassium hydroxide using 0.5 mL of
the chromatogram obtained with reference solution (b) phenolphthalem solution R l as indicator. Titrate the solution
(0.05 per cent).
immediately while still hot.
Loss on drying (2.2.52) Peroxide value (2.5.5, Method A)
Maximum 0.5 per cent, determined on 0.500 g by drying in Maximum 5.0.
an oven at 105 °C.
Unsaponifiable m atter (2.5.7)
ASSAY Maximum 1.0 per cent, determined on 5.0 g.
Dissolve 0.100 g in ethanol (96 per cent) R and dilute to
Alkaline impurities
100.0 mL with the same solvent Dilute 2.0 mL of this
Dissolve by gentle heating 2.0 g in a mixture of 1.5 mL of
solution to 100.0 mL with ethanol (96 per cent) R. Measure
ethanol (96 per cent) R and 3 mL of toluene R. Add 0.05 mL
the absorbance (2.2.25) at the absorption maximum at
of a 0.4 g/L solution of bromophenol blue R in ethanol
237 nm.
(96 per cent) R. Not more than 0.4 mL of 0.01 M hydrochloric
Calculate the content of C23H 30O6 taking the specific add is required to change the colour to yellow.
absorbance to be 395.
Composition of fatty acids (2.422, Method A)
STORAGE Use the mixture of calibrating substances in Table 2.4.22.-3.
Protected from light. Column:
IMPURITIES — material: fused silica;
Specified impurities A — size. 1= 25 m, 0 = 0.25 mm;
— stationary phase: poly(tyanopropyl)sibxane R (film thickness
0.2 jxm).
Carrier gas hdium for chromatography R.
2016 Cresol 1-657
Flow rate 0.65 mL/min. B. Add bromine water. A pale yellow flocculent precipitate is
Split ratio 1:100. produced.
Temperature: TESTS
— column: 180 °C for 35 min; Acidity
— injection port and detector. 250 °C. A 2.0% w/v solution is neutral to bromocresolpurple solution.
Detection Flame ionisation. Distillation range
Composition of thefatty-acid fraction of the oU: Not more than 2% v/v distils below 188° and not less than
— saturatedfatty acids of chain length less than Crf. maximum 80% v/v distils between 195° and 205°, Appendix V C.
0.2 per cent; Weight p er mL
— myrisdc acid: maximum 1.0 per cent; 1.029 to 1.044 g, Appendix V G.
— palmitic acid: 19.0 per cent to 26.0 per cent;
— stearic acid: 68.0 per cent to 80.0 per cent; Hydrocarbons
— oleic acid and isomers: maximum 4.0 per cent; Place 5 0 mL in a 5 0 0 mL round-bottomed flask, add
150 mL of 5 m sodium hydroxide and 3 0 mL of water and mix
— linoleic acid and isomers: maximum 1.0 per cent;
— arackidic acid: maximum 1.0 per cent; thoroughly. Connect the flask to a splash-bulb and air-
— behenic acid: maximum 1.0 per cent; condenser about 6 0 cm long, with the end of the air-
—' Ugnoceric acid: maximum 0.5 per cent. condenser fitting closely into the neck of a 2 5 0 mL pear-
shaped separating funnel and passing well into the separating
Nickel funnel, which has a cylindrical graduated portion above the
Maximum 1 ppm. stopcock. Fill the graduated portion of the separating funnel
Atomic absorption spectrometry (2.2.23, Method II). with water. Distil rapidly until 75 mL of distillate has been
Test solution Introduce 5.0 g into a platinum or silica crucible collected, cooling the separating funnel in running water if
tared after ignition. Cautiously heat and introduce into the necessary. Allow the separating funnel to stand in a vertical
substance a wick formed from twisted ashless filter paper. position until separation is complete and draw off the
Ignite the wick. When the substance ignites, stop heating. aqueous liquid into a titration flask for use in the test for
After combustion, ignite in a muffle furnace at about Volatile bases.
600 ± 50 °C. Continue the incineration until white ash is Allow the separating funnel to stand for a few minutes,
obtained. After cooling, take up the residue with 2 quantities, measure the volume of hydrocarbon oil in the graduated
each of 2 mL, of dilute hydrochloric acid R and transfer into a portion and warm, if necessary, to keep the oil in the liquid
25 mL graduated flask. Add 0.3 mL of nitric acid R and state. Subtract the volume of volatile bases in the
dilute to 25.0 mL with distilled water R. hydrocarbon oil, as determined in the following test
Reference solutions Prepare 3 reference solutions by adding Not more than 0.15% v/v of hydrocarbon oil is present.
1.0 mL, 2.0 ml. and 4.0 ml. of nickel standard solution Volatile bases
(0.2 ppm Ni) R to 2.0 mL portions of the test solution, To the aqueous liquid reserved in the test for Hydrocarbons
diluting to 10.0 mL with distilled water R. add any aqueous liquid still remaining in the separating
Source Nickel hollow-cathode lamp. funnel and neutralise, if necessary, with 0 . 1 m hydrochloric acid
Wavelength 232 am. using phenolphthalein solution R l as indicator. Titrate with
1m hydrochloric acid FS using methyl orange solution as
Atomisation device Graphite furnace.
indicator. Wash the oil from the separating funnel into the
Carrier gas argon R.
titration flask with water and again titrate with 1m hydrochloric
STORAGE acid VS. From the volume of additional 1m hydrochloric acid
Protected from light. FS, calculate the proportion of volatile bases in the
Ph Eur hydrocarbon oil. From the total volume of 1m hydrochloric
add VS used in both titrations calculate the volume of
volatile bases in the substance being examined. Each mL of
1m hydrochloric acid FS is equivalent to 0.080 mL of volatile
bases. Not more than 0.15% v/v of volatile bases is present.
Cresol Hydrocarbons and volatile bases
The sum of the contents of hydrocarbon oil and volatile
Action and use bases, as determined in the tests for Hydrocarbons and for
Antiseptic; antimicrobial preservative. Volatile bases, does not exceed 0.25% v/v.
DEFINITION Sulfur compounds
Cresol is a mixture of cresols and other phenols obtained Place 20 mL in a small conical flask and over the mouth of
from coal tar. the flask fix a piece of filter paper moistened with a 10% w/v
solution of lead(n) acetate. Heat the flask on a water bath for
CHARACTERISTICS 5 minutes. Not more than a light yellow colour is produced
An almost colourless to pale brownish yellow liquid. on the filter paper.
Almost completely soluble in 50 volumes of water, freely Non-volatile m atter
soluble in ethanol (96%), in ether and in fixed and volatile
When evaporated on a water bath and dried at 105°, leaves
oils. not more than 0 . 1% w/v of residue.
IDENTIFICATION STORAGE
Shake 0.5 mL with 300 mL of water and filter. The filtrate
Cresol should be protected from light. It darkens with age or
complies with the following tests.
on exposure to light.
A Add inm(m) chloride solution R l. A transient blue colour is
produced.
1-658 Crude Cresol 2016
>** ★
★ .★**
Crude Cresol ★ ★ Croscarmellôse Sodium *
* *
*
***** *****
(Ph. Eur. monograph 1628) (Ph. Eur. monograph 0985)
★ ★ TESTS
Crospovidone ★ ★ Peroxides
(Ph. Eux. monograph 0892) ***** Type A maximum 400 ppm expressed as H 2O 2; type B:
maximum 1000 ppm expressed as H 2O2.
Suspend 2.0 g in 50 mL of water R. To 25 mL of this
suspension add 2 mL of titanium trichloride-sulfuric add
reagent R. Allow to stand for 30 min and filter.
The absorbance (2.2.25) of the filtrate, measured at 405 nm
using a mixture of 25 mL of a filtered 40 g/L suspension of
the substance to be examined and 2 mL of a 13 per cent VIV
(CeHçNO)^ 9003-39-8 solution of sulfuric acid R as the compensation liquid, has a
maximum of 0.35.
Action and use For type B use 10 mL of the suspension and dilute to 25 mL
Excipient in pharmaceutical products. with water R for the test.
PhEir.
W ater-soluble substances
Maximum 1.5 per cent
DEFINITION Place 25.0 g in a 400 mL beaker, add 200 mL of water R
Cross-linked homopolymer of 1-ethenylpyrrolidin-2-one. and stir for 1 h using a magnetic stirrer. Transfer the
Content suspension to a 250.0 mL volumetric flask, rinsing with
11.0 per cent to 12.8 per cent of N (AT 14.01) (dried water i?, and dilute to volume with the same solvent. Allow
substance). the bulk of the solids to setde. Filter about 100 mL of the
2 types of crospovidone are available, depending on the almost clear supernatant through a membrane filter (nominal
particle size: type A and type B. pore size 0.45 pm), protected by superimposing a membrane
filter (nominal pore size 3 pm). While filtering, stir the liquid
CHARACTERS
above the membrane filter manually or by means of a
Appearance mechanical stirrer, taking care not to damage the membrane
Hygroscopic, white or yellowish-white powder or flakes. filter. Transfer 50.0 mL of the clear filtrate to a tared
Solubility 100 mL beaker, evaporate to dryness and dry at 105-110 °C
Practically insoluble in water, in ethanol 96 per cent and in for 3 h. The residue weighs a maximum of 75 mg.
methylene chloride. Im purity A
IDENTIFICATION Liquid chromatography (2.2.29).
A. Infrared absorption spectrophotometry (2.2.24). Test solution Suspend 1.250 g in 50.0 mL of methanol R and
Comparison crospovidone CRS. shake for 60 min. Leave the bulk to settle and filter through
B. Suspend 1 g in 10 mL of water R , add 0.1 mL of 0.05 M a membrane filter (nominal pore size 0.2 pm).
iodine and shake for 30 s. Add 1 mL of starch solution R and Reference solution (a) Dissolve 50 mg of 1-vinylpyrrolidm-2-
shake. No blue colour develops within 30 s. one R (impurity A) in methanol R and dilute to 100.0 mL
C. To 10 mL of water R, add 0.1 g and shake. A suspension with the same solvent. Dilute 1.0 mL of the solution to
is formed and no clear solution is obtained within 15 min. 100.0 mL with methanol R. Dilute 5.0 mL of this solution to
100.0 mL with the mobile phase.
D. The analytical sieves must be clean and dry. For this purpose
the sieves are washed in hot water and allowed to dry overnight in Reference solution (b) Dissolve 10 mg of l-vinylpyrrolidin-2-
a drying cabinet at 105 °C. one R (impurity A) and 0.50 g of vinyl acetate R in
methanol R and dilute to 100 mL with the same solvent.
Place 20 g (dried substance) in a 1000 mL conical flask, add
Dilute 1.0 mL of the solution to 100.0 mL with the mobile
500 mL of water R and shake the suspension for 30 min.
phase.
Pour the suspension through a 63 jam analytical sieve,
previously tared, and rinse the sieve with water R until the Precolumn:
filtrate is clear. Dry the sieve and sample residue at 105 °C — size. I = 0.025 m, 0 = 4 mm;
for 5 h in a drying cabinet without circulating air. Cool in a — stationary phase. octadecylsUyl silica gelfor chromatography R
desiccator for 30 min and weigh- (5 pm).
Column:
Calculate the percentage sieving residue (fraction of sample
particles having a diameter of more than 63 pm), using the — size. I = 0.25 m, 0 = 4 mm;
following expression: — stationary phase. octadecylsUyl silica gelfor chromatography R
(5 pm);
m i —m 3 — temperature. 40 °C.
x 100 Mobile phase acetomtrüe R , water R (10:90 VIV).
m2
Flow rate 1 mL/min.
m\ — mass of the sieve and sample residue, after drying
Detection Spectrophotometer at 235 nm.
for 5 h, in grams; Injection 50 pL. After each injection of die test solution, wash
m2 = initial mass of the sample, in grains; the precolumn by passing the mobile phase backwards, at the
m3 = mass of the sieve, in grams. same flow rate as applied in the test, for 30 min.
If the sieving residue fraction is more than 15 per cent, the System suitability:
substance is classified as type A; if the sieving residue fraction — resolution: minimum 2.0 between the peaks due to
is less than or equal to 15 per cent, the substance is classified impurity A and vinyl acetate in the chromatogram
as type B. obtained with reference solution (b);
2016 Crotamiton 1-661
— repeatability: maximum relative standard deviation of criteria. In such cases, a cross-reference to the tests described in the
2.0 per cent after 6 injections of reference solution (a). mandatory part is included m the Functionality-related
Calculation ofpercentage content. characteristics section. Control of the characteristics can contribute
— for impurity A, use the concentration of impurity A in to the quality of a medicinal product by improving the consistency
reference solution (a). of the manufacturing process and the performance of the medicinal
Limit: product during use. Where control methods are cited, they are
— impurity A: maximum 10 ppm. recognised as being suitable for the purpose, but other methods can
also be used. Wherever resultsfor a particular characteristic are
Heavy m etals (2.4.8) reported, the control method must be indicated.
Maximum 10 ppm.
The following characteristics may be relevantfor crospovidone used
2.0 g complies with test D. Prepare the reference solution as disintegrant
using 2 mL of lead standard solution (10 ppm Pb) R.
Hydration capacity
Loss on drying (2.2.32) Introduce 2.0 g into a 100 mL centrifuge tube and add
Maximum 5.0 per cent, determined on 0.500 g by drying in 40 mL of water R. Shake vigorously until a suspension is
an oven at 105 °C. obtained. Shake again 5 min and 10 min later, then
Sulfated ash (2.4.14) centrifuge for 15 min at 750 g. Decant the supernatant and
M axim um 0.1 per cent, determined on 1.0 g. weigh the residue. The hydration capacity is the ratio of the
ASSAY mass of the residue to the initial mass of the sample. It is
Place 0.100 g of the substance to be examined (m mg) in a typically 3 to 9.
combustion flask and add 5 g of a mixture of 1 g of copper Particle-size distribution (2.9.31)
sulfate R, 1 g of titanium dioxide R and 33 g of dipotassium Powder flow (2.9.36)
sulfate R, and 3 glass beads. Wash any adhering particles The following characteristic may be relevant for crospovidone used
from the neck into the flask with a small quantity of water R. as suspension stabiliser.
Add 7 mL of sulfuric acid R, allowing it to run down the
Settling volume
inside wall of the flask. Gradually heat the flask until the
Introduce 10 g into a 100 mL graduated cylinder and add
solution has a clear, yellowish-green colour, and the inside
90 mL of water R. Shake vigorously. Dilute to 100 mL with
wall of the flask is free from carbonised material, and then
water R, washing the powder residues from the walls of the
heat for a further 45 min. After cooling, cautiously add
cylinder. Allow to stand for 24 h, then read the volume of
20 mL of water R, and connect the flask to the distillation
the sediment. It is typically greater than 60 m L
apparatus, which has been previously washed by passing
steam through it. To the absorption flask add 30 mL of a PhEur
40 g/L solution of boric acid R, 0.15 mL of bromocresol green-
methyl red solution R and sufficient water R to immerse the
lower end of the condenser tube. Add 30 mL of strong sodium
hydroxide solution R through a funnel, cautiously rinse the ★ ★.
Crotamiton ★ ★
funnel with 10 mL of water R, immediately close the clamp
attached to the rubber tube, then start the distillation with (Ph Eta", monograph 1194) *****
steam to obtain 80-100 mL of distillate. Remove the
absorption flask from the lower end of the condenser tube, H3c
rinsing the end part with a small quantity of water R, and 'I ?Ha
h3^ \ ^ N^ and (Z)-isomer
titrate the distillate with 0.025 M sulfuric acid until the colour
of the solution changes from green through pale greyish-blue
to pale greyish red-purple. Carry out a blank determination
and make any necessary correction. CjaHnNO 203.3 483-63-6
1 mL of 0.025 M sulfuric acid is equivalent to 0.700 mg of N.
Action and use
STORAGE
Acaridde.
In an airtight container.
Preparations
LABELLING Crotamiton Cream
The label states the type of crospovidone (type A or type B).
Crotamiton Lotion
IMPURITIES
PhEtr.
ch 2
r DEFINITION
JV-Ethyl-N-(2-methylphenyI)but-2-enamide.
C r ° Content
— sum of the (E)- and (Z)-isomers: 96.0 per cent to
102.0 per cent;
A. 1-ethenylpyrrolidin-2-one (1-vinylpyrrolidin-2-one).
— (Z)-isomer, m axim um 15.0 per cent.
FUNCTIONALITY-RELATED CHARACTERISTICS
CHARACTERS
This section provides information on characteristics that are
Appearance
recognised as being relevant control parameters for one or more
Colourless or pale yellow, oily liquid.
functions of the substance when used as an excipient (see chapter
5.15). Some of the characteristics described in the Functionality- Solubility
related characteristics section may also be present in the mandatory Slightly soluble in water, misdble with ethanol (96 per cent).
part of the monograph since they also represent mandatory quality At low temperatures it may partly or completely solidify.
1-662 Crotamiton 2016
Maximum 100 ppm. peaks due to the (Z)- and (£)-isomers in the
chromatogram obtained with reference solution (c)
Boil 5.0 g under a reflux condenser for 1 h with 25 mL of
(0.02 per cent).
ethanol (96 per cent) R and 5 mL of a 200 g/L solution of
sodium hydroxide R. Cool, add 5 mL of water R and shake Sulfated ash (2.414)
with 25 mL of ether R. Dilute the lower layer to 20 mL with Maximum 0.1 per cent, determined on 1.0 g.
water R-, add 5 mL of nitric acid R, dilute to 50 mL with ASSAY
water R and add 1 mL of a freshly prepared 50 g/L solution Liquid chromatography (2.2.29) as described in the test for
of silver nitrate R. Any opalescence in the solution is not more related substances with the following modification.
intense than that in a mixture of 1 mL of a freshly prepared Injection Test solution (b) and reference solution (a).
50 g/L solution of silver nitrate R and a solution prepared by
diluting 5 mL of a 200 g/L solution of sodium hydroxide R to Calculate the percentage content of C 13H 17NO from the sum
20 mL with water R and adding 1.5 mL of 0.01 M of the areas of the peaks due to the (2)- and (J5)-isomers in
the chromatograms obtained. Calculate the content of the
2016 Cyanocobalamin 1-663
DEFINITION SY ST E M S U IT A B IL IT Y
Cyclizine is l-benzhydryl-4-methyipiperazine. It contains not Inject solution (3) 6 times. The rdative standard deviation of
less than 98.5% and not more than 101.0% of C 18H 22N 2, each of the areas of the 3 prinripal peaks is not more than
calculated with reference to the dried substance. 5.0%.
CHARACTERISTICS The test is not valid unless, in the chromatogram obtained
A white or creamy white, crystalline powder. with solution (3);
Practically insoluble in water. It dissolves in most organic the peak-to-vaUey ratio between methanol and
solvents and in dilute adds. 1-methylpiperazine (impurity A) is at least 50;
IDENTIFICATION the resolutionfactor between diphenylmethanol (impurity B)
A. The infrared absorption spectrum, Appendix II A, is and cyclizine is at least 18.
concordant with the reference spectrum of cyclizine (RS 075). L IM IT S
B. Melting point, about 107°, Appendix V A In the chromatogram obtained with solution (1):
the area of the peak corresponding to 1-methylpiperazine
(impurity A) is not greater than the peak corresponding to
1-methylpiperazine in solution (3) (0.5%);
2016 Cyclizine Hydrochloride 1-665
the area of the peak corresponding to diphenylmethanol Test solution (b)Dilute 10.0 mL of test solution (a) to
(impurity B) is not greater than the peak corresponding to 100.0 mL with a 5 g/L solution of sulfuric acid R.
diphenylmethanol in solution (3) (0.5%); Spectral range 240-350 nm for test solution (a); 210-240 nm
the area of any other secondary peak is not greater than the for test solution (b).
area of the principal peak in the chromatogram obtained with Resolution (2.2.25): minimum 1.7.
solution (2) (0.1%); Absorption maxima At 258 nm and 262 nm for test
the Siam of the areas of all secondary peaks is not greater than solution (a); at 225 nm for test solution (b).
10 times the area of the principal peak in the chromatogram Absorbance raâo A262/A258 = 1.0 to 1.1.
obtained with solution (2) (1.0%).
Specific absorbance at the absorption maximum at 225 nm
Disregard any peak with an area less than 0.5 times that of 370 to 410 for test solution (b).
the principal peak in the chromatogram obtained with
solution (2) (0.05%). B. Infrared absorption spectrophotometry (2.2.24).
Comparison cyclizine hydroçjdoride CRS.
Loss on drying
When dried to constant weight at 80°, loses not more than C. Thin-layer chromatography (2.2.27).
1.0% of its weight. Use 1 g. Test solution Dissolve 10 mg of the substance to be examined
Sulfated ash in methanol R and dilute to 10 mL with the same solvent.
Not more than 0.1%, Appendix IX A. Reference solution Dissolve 10 mg of cyclizine
hydrochloride CRS in methanol R and dilute to 10 mL with the
ASSAY
same solvent.
Carry out Method I for non-aqueous titration,
Plate TLC silica gel GF254 plate R.
Appendix VIII A, using 0.1 g and determining the end point
potentiometrically. Each mL of 0.1m perchloric add P'S is Mobile phase concentrated ammonia R, methanol R, methylene
equivalent to 13.32 mg of C18H22N2. chloride R (2:13:85 V/V/V).
Application 20 |iL.
Development Over 2/3 of the plate.
Drying In air for 30 min.
*****
Cyclizine Hydrochloride * *
Detection Expose to iodine vapour for 10 min,
*•+ ** Restdts The prindpal spot in the chromatogram obtained with
(Ph. Eur. monograph 1092) **+
the test solution is similar in position, colour and size to the
principal spot in the chromatogram obtained with the
reference solution.
D. Dissolve 0.5 g in 10 mL of ethanol (60 per cent VIV) R,
Ha heating if necessary. Cool in iced water. Add 1 mL of dilute
sodium hydroxide solution R and 10 mL of water R. Filter,
wash the predpitate with water R and dry at 60 °C at a
pressure not exceeding 0.7 kPa for 2 h. The melting point
(2.2.14) is 105 °C to 108 °C.
Ci8H23ClN2 302.8 305-25-3
E. It gives reaction (a) of chlorides (2.3.1).
Action and use TESTS
Histamine Hi receptor antagonist; antihistamine. pH (2.2.3)
Preparations 4.5 to 5.5.
Cyclizine Tablets Dissolve 0.5 g in a mixture of 40 volumes of ethanol
Dipipanone and Cyclizine Tablets (96 per cent) R and 60 volumes of carbon dioxide-free water R
and dilute to 25 mL with the same mixture of solvents.
PhEir______________________________________________________
Related substances
DEFINITION Gas chromatography (2.2.28). Prepare the solutions immediately
1-(Diphenylmethyl)-4-methylpiperazine hydrochloride. before use. _
Content Test solution Dissolve 0.250 g of the substance to be
98.5 per cent to 101.0 per cent (dried substance). examined in 4.0 mL of methanol R and dilute to 5.0 mL with
1 M sodium hydroxide.
CHARACTERS
Appearance Reference solution (a) Dissolve 25 mg of the substance to be
White or almost white, crystalline powder. examined in 10.0 mL of methanol R. Dilute 1.0 mL of this
solution to 50.0 mL with methanol R.
Solubility
Reference solution (b) Dissolve 5 mg of the substance to be
Sligjhtly soluble in water and in ethanol (96 per cent).
examined, 5.0 mg of cyclizine impurity A CRS and 5.0 mg of
IDENTIFICATION cyclizine impurity B CRS in methanol R and dilute to 20.0 mL
First identification B, E. with the same solvent.
Second identification A , C, D, E. Column:
A. Ultraviolet and visible absorption spectrophotometry — material: fused silica;
(2.2.25). — size: I = 25 m, 0 = 0.33 mm;
Test solution (a) Dissolve 20.0 mg in a 5 g/L solution of — stationary phase: poly(dimethyl) (diphenyl)sUoxane R (film
sulfuric addR and dilute to 100.0 mT. with the same add thickness 0.50 pm).
solution. Carrier gas helium for chromatography R.
1-666 Cyclopenthiazide 2016
Time Temperature
(min) CC)
Column 0 -1 4 100 ->240 B. diphenylmethanol (benzhydrol).
___________________________________________________________PhEur
14-16 240 -» 270
16-30 270
0.0050% w/v. After removal of the plate, dry it in a current Test solution Dissolve 10 mg of the substance to be examined
of air and reveal the spots by Method. I. Any secondary spot in in 5 mL of ethanol (96 per cent) R.
the chromatogram obtained with solution (1) is not more Reference solution Dissolve 10 mg of cyclopentolate
intense rhan the spot in the chromatogram obtained with hydrochloride CRS in ethanol (96 per cent) R and dilute to
solution (2). 5 mL with the same solvent.
Loss on drying Plate TLC silica gel plate R.
When dried to constant weight at 105°, loses not more than Mobile phase concentrated ammonia R, water R, butyl acetate R,
0.5% of its weight Use 1 g. 2-propanol R (5:15:30:50 VIVIV/V).
Sulfated ash Application 10 pL.
Not more than 0.1%, Appendix IX A.
Development Over 2/3 of the plate.
ASSAY Drying In air.
Dissolve 0.5 g in 50 mL of butylamine and carry out Detection Spray with alcoholic solution of sulfuric acid R and
Method II for rum-aqueous titration, Appendix VHI A, using heat at 120 °C for 30 min; examine in ultraviolet light at
0.1m tetrabutylammonium hydroxide KS as titrant and 365 nm.
magneson solution as indicator; titrate to a pure blue end
Result The principal spot in the chromatogram obtained with
point. Each mL of 0.1m tetrabutylammonium hydroxide FS is
the test solution is similar in position, fluorescence and size
equivalent to 18.99 mg of C 13H 18CIN3O4S2.
to the principal spot in the chromatogram obtained with the
reference solution.
D. It gives reaction (a) of chlorides (2.3.1).
TESTS
Cyclopentolate Hydrochloride ***** pH (2.2.3)
*. *
(Ph. Eur. monograph 1093) * 4.5 to 5.5.
Dissolve 0.2 g in carbon dioxide-free water R and dilute to
20 mL with the same solvent.
Related substances
Liquid chromatography (2.2.29). Prepare the solutions
immediately before use.
Test solution Dissolve 20 mg of the substance to be examined
inwater R and dilute to 20.0 mL with the same solvent
C17H26d N 0 3 327.8 5870-29-1 Reference solution (a) Dilute 1.0 mL of the test solution to
100.0 mL with water R. Dilute 5.0 mL of this solution to
Action and use 10.0 mL with water R.
Anticholinergic. < Reference solution (b) Dissolve 10 mg of cyclopentolatefor
Preparation system suitability CRS (con taining impurity C) in water R and
Cyclopentolate Eye Drops dilute to 10.0 mL with the same solvent.
Column:
PhEur__________________________________________________________
— size. I = 0.125 m, 0 = 4.0 mm;
DEFINITION — stationary phase, spherical end-capped hexylsUyl silica gelfor
2-(Dimethylamino)ethyl (2RS)- chromatography R (5 pm).
(1-hydroxycyclopentyl) (phenyl) acetate hydrochloride. Mobile phase Dissolve 0.66 g of ammonium phosphate R in
Content water R, adjust to pH 3.0 with phosphoric add R and dilute to
98.5 per cent to 101.5 per cent (dried substance). 1000 mL with water R; mix and filter; mix 55 volumes of this
solution and 45 volumes of acetonitrüe R l.
CHARACTERS
Appearance Flow rate 1.0 ml /min.
White or almost white, crystalline powder. Detection Spectrophotometer at 220 nm.
Solubility Injection 20 pi.
Very soluble in water, freely soluble in ethanol (96 per cent). Run time 2.5 times the retention time of cyclopentolate.
It shows polymorphism (5.9). Identification of impurities Use the chromatogram supplied
Limits:
★*★
Cyclophosphamide ★ ★
— correctionfactor, for the calculation of content, multiply the ★ ★
peak area of impurity C by 2.0; (Ph. Eur. monograph 0711) *****
— impurity C: not more than the area of the principal peak
in the chromatogram obtained with reference solution (a) Cl
(0.5 per cent);
— unspecified impurities: for each impurity, not more than and enantiomer , H20
0.2 times the area of the principal peak in the
chromatogram obtained with reference solution (a)
(0.10 per cent);
— total: not more than twice the area of the principal peak in CyHjgCysr^PjHzO 279.1 6055-19-2
the chromatogram obtained with reference solution (a)
(1.0 per cent); Action and use
— disregard limit. 0.1 times the area of the principal peak in Cytotoxic alkylating agent.
the chromatogram obtained with reference solution (a) Preparations
(0.05 per cent). Cydophosphamide Injection
Loss on drying (2.2.32) Cyclophosphamide Oral Solution
Maximum 0.5 per cent, determined on 1.000 g by drying in Cydophosphamide Tablets
an oven at 105 °C for 4 h.
Sulfa ted ash (2.4.14) PhEur______________________________
Maximum 0.1 per cent, determined on 1.0 g. DEFINITION
ASSAY Cyclophosphamide contains not less than 98.0 per cent and
Dissolve 0.250 g in a mixture of 1.0 mL of 0.1 M hydrochloric not more than the equivalent of 102.0 per cent of (2RS)-
add and 50 mL of ethanol (96 per cent) R. Cany out a AyV-bis(2-chloroethyl)tetrahydro-2ii-l,3,2-oxa2aphosphorin-
potentiometric titration (2.2.20), using 0.1 M sodium 2-amine 2-oxide, calculated with reference to the anhydrous
hydroxide. Read the volume added between the 2 points of substance.
inflexion. CHARACTERS
1 mL of 0.1 M sodium hydroxide is equivalent to 32.79 mg of A white or almost white, crystalline powder, soluble in water,
C17H 26C1N03. freely soluble in alcohol.
IMPURITIES IDENTIFICATION
Specified impurities C First identification B.
Other detectable impurities (the following substances would, if Second identification A, C, D.
present at a sufficient level, be detected by one or other of A. Determine the mdting point (2.2.14) of the substance to
the tests in the monograph. They are limited by the general be examined. Mix equal parts of the substance to be
acceptance criterion for other/unspecified impurities and/or examined and cyclophosphamide CRS and determine the
by the general monograph Substances for pharmaceutical use melting point of the mixture. The difference between the
(2034). It is therefore not necessary to identify these melting points (which are about 51 °C) is not greater than
impurities for demonstration of compliance. See also 5.10. 2 °C.
Control of impurities in substances for pharmaceutical use): A, B.
B. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
cydophosphamide CRS.
C. Examine the chromatograms obtained in the test for
rdated substances. The principal spot in the chromatogram
obtained with test solution (b) is similar in position, colour
and size to the prindpal spot in the chromatogram obtained
with reference solution (a).
D. Dissolve 0.1 g in 10 m l. of water R and add 5 mL of
silver nitrate solution R l; the solution remains dear. Boil, a
white predpitate is formed which dissolves in concentrated
ammonia R and is repredpitated on the addition of dilute
nitric add R.
Related substances ★ *★
Cyproheptadine Hydrochloride ★ ★
Examine by thin-layer chromatography (2.2.27), using silica ★ ★
gel G R as the coating substance. (PA. Eur. monograph 0817) *****
Test solution (a) Dissolve 0.10 g of the substance to be
examined in alcohol R and dilute to 5 mL with the same
solvent.
Test solution (b) Dilute 1 mL of test solution (a) to 10 mL . HCI , 1’/,H20
with alcohol R.
Reference solution (a) Dissolve 10 mg of cyclophosphamide CRS
in alcohol R and dilute to 5 mL with the same solvent.
Reference solution (b) Dilute 0.1 mL of test solution (a) to
10 ml, with alcohol R. CaHaaON.l&HaO 350.9 41354-29-4
Apply separately to the plate 10 |iL of each solution. Develop
Action and use
over a path of 15 cm using a mixture of 2 volumes of
Histamine Hx receptor antagonist; antihistamine.
anhydrous formic acid R, 4 volumes of acetone R, 12 volumes
of water R and 80 volumes of methyl ethyl ketone R. Dry the Preparation
plate in a current of warm air and heat at 110 °C for 10 min. Cyproheptadine Tablets
At the bottom of a chromatographic tank, place an PhEir.
evaporating dish containing a 50 g/L solution of potassium
permanganate R and add an equal volume of hydrochloric DEFINITION
acid R. Place the plate whilst still hot in the tank and close 4-(5H-Dibenzo [a,d[ [7] annulen-5-ylidene)-1 -methylpiperidine
the tank. Leave the plate in contact with the chlorine gas for hydrochloride sesquihydrate.
2 min. Withdraw the plate and place it in a current of cold Content
air until the excess of chlorine is removed and an area of 98.5 per cent to 101.0 per cent (anhydrous substance).
coating below the points of application gives at most a very
faint blue colour with a drop of potassium iodide and starch CHARACTERS
solution R. Avoid prolonged exposure to cold air. Spray with Appearance
potassium iodide and starch solution R and allow to stand for White or slightly yellow, crystalline powder.
5 mm. Any spot in the chromatogram obtained with test Solubility
solution (a), apart from the principal spot, is not more Slightly soluble in water, freely soluble in methanol, sparingly
intense than the spot in the chromatogram obtained with soluble in ethanol (96 per cent).
reference solution (b) (1.0 per cent). Disregard any spot IDENTIFICATION
remaining at the point of application.
A. Infrared absorption spectrophotometry (2.2.24).
Chlorides (2.4.4) Comparison cyproheptadine hydrochloride CRS.
Dissolve 0.15 g in water R and dilute to 15 mL with the
same solvent. The freshly prepared solution complies with B. A saturated solution gives reaction (b) of chlorides (2.3.1).
Mobile phase:
— mobile phase A: dissolve 6.12 g of potassium dihydrogen
phosphate R in 900 mL of water R, adjust to pH 4.5 with
phosphoric add R and dilute to 1000 mL with water R',
mix 60 volumes of this solution and 40 volumes of
acetonitrUe for chromatography R;
— mobile phase B: dissolve 6.12 g of potassium dihydrogen
A. 5H-dibenzo[ajii] [7]annulene (dibenzocycloheptene),
phosphate R in 900 mL of water R, adjust to pH 4.5 with
phosphoric add R and dilute to 1000 mL with water R\
mix 40 volumes of this solution and 60 volumes of
acetonitrile for chromatography R',
10.0 - 10.1 100 -»0 0 ^ 100 B. 10,11 -dihydro- 5//-dibenzo [a3d\[7] annul en-5-one
(dibenzosuberone),
10.1 - 35 0 100
G. 6 ß-chloro-7a-hydroxy-3,20-dioxo-1ß,2 ß-dihydro-3'H-
cyclopropa[1,2]pregna-1,4-dien-17-yl acetate,
A. 3,20-dioxo-1ß,2ß-dihydro-3 '/f-cydopropa [1,2]pregna-
l,4,6-trien-17-yl acetate.
H. 3,20-dioxopregna-l,4-dien-17-yl acetate,
B. 6-methoxy-3,20-dioxo-lß,2ß-dihydro-3'H-
cyclopropa [1,2] pregna-1,4,6-trien-17-yl acetate,
I. 6-chloro-3,20-dioxopregna-1,4,6-trien-17-yl acetate
(delmadinone acetate),
C. 6-chloro-1a-(chloromethyi)-3,20-dioxopregna-4,6-dien-17-
yl acetate,
J. 6oc,7a-epoxy-3,20-dioxo-lß,2ß-dihydro-3 'H-
cyclopropa[1,2]pregna-1,4-dien-17-yl acetate.
___________________________________________________________ PhEur
D. la-(chloromethyl)-3,6,20-trioxopregn-4-en-17-yl acetate,
H NHj
H S^ X ■ HCI , H p
C02H
DEFINITION
(2i?)-2-Ammo-3-sulfanylpropanoic add hydrochloride
monohydrate.
Fermentation product, extract or hydrolysate of protein.
F. 6-chloro-17-hydroxy-1ß, 2 ß-dihydro-3 'H- Content
cyclopropa[l,2]pregna-l,4,6-txiene-3,20-dione, 98.5 per cent to 101.0 per cent (dried substance).
2016 Cysteine Hydrochloride 1-673
A m m o n iu m
Amino add analysis (2.2.56) as described in the test for
Cystine ;****,
★ ★
ninhydrin-positive substances with the following (Ph. Eur. monograph 0998) **
modifications.
Injection Test solution, reference solution (d) and blank H nh 2
Column'.
20 mL vial. Using a 10 pL syringe, inject 10 pL of
— size: I = 0.25 m, 0 = 4.6 mm; solution A and 10 pL of a 10 g/L solution of triethylamine R
— stationary phase: octadecylsUyl silica gelfor into the vial.
chromatography R (5 jam). Column:
— material: fused silica;
Mobile phase 15.63 g/L solution of glacial acetic add R
— sizer. I = 30.0 m, 0 = 0.53 mm;
containing 2.33 g/L of sodium dioctyl sulfosucdnate R As the
— stationary phase: base-deactivated polyethylenegfycol R (film
mobile phase contains sodium dioctyl sulfosucdnate, it must
thickness 1.0 pm).
be freshly prepared every day, and the column must be
flushed with a mixture of equal volumes of methanol R and Carrier gas helium for chromatography R.
water R, after all tests have been completed or at the end of Flow rate 13 mL/min.
the day, for at least 2 h. Split ratio 1:1.
Flow rate 1.2 mL/min. Static head-space conditions that may be used:
Detection Spectrophotometer at 254 nm. — equilibration temperature: 60 °C;
Action and use Reference solution (b) Dilute 1.00 mL of reference solution (a)
Low molecular weight heparin. to 50.0 mL with water R.
Before preparing reference solutions (c), (d) and (e), rinse all
Preparation
pipettes with reference solution (b).
Dalteparin Sodium Injection
2016 Danaparoid Sodium 1-679
Reference solution (c) Dilute 1.00 mL of reference solution (b) Reference solution (b) Prepare a 11.4 |ig/mL solution of boric
to 100.0 mT. with water R (corresponding to 1 ppm of nitrite acid R in a 1 per cent V/V solution of nitric acid R in waterfor
in the test sample). chromatography R (STD^d).
Reference solution (d) Dilute 3.00 mL of reference solution (b) Reference solution (c) Dissolve 0.2500 g of a reference
to 100.0 mL with water R (corresponding to 3 ppm of nitrite dalteparin sodium with no detectable boron in about 2 mL of
in the test sample). waterfor chromatography R, add 100 (iL of nitric acid R and
Reference solution (e) Dilute 5.00 mL of reference solution (b) dilute to 10.00 mL with the same solvent (STD0).
to 100.0 mL with water R (corresponding to 5 ppm of nitrite Reference solution (d) Dissolve 0.2500 g of a reference
in the test sample). dalteparin sodium with no boron detected in about 2 mL of
The chromatographic procedure may be carried out using: a 1 per cent V/V solution of nitric acid R in waterfor
— a column 0.125 m long and 4.3 mm in internal diameter chromatography R, add 10 (iL of a 5.7 mg/mL solution of
packed with a strong anion-exchange resin; boric add R and dilute to 10.00 mL with the same solvent
— as mobile phase at a flow rate of 1.0 mL/min a solution (STDi). This solution contains l pg/mL of boron.
consisting of 13.61 g of sodium acetate R dissolved in Calculate the content of boron in the substance to be
water R, adjusted to pH 4.3 with phosphoric acid R and examined, using the following correction factor
diluted to 1000 mL with water R'}
— as detector an appropriate electrochemical device with the j = (STDi - STDo) x 2
following characteristics and settings: a suitable working (STDcai —blank)
electrode, a detector potential of + 1.00 V versus
Ag/AgCl reference electrode and a detector sensitivity of Loss on drying (2.2.32)
0.1 jiA full scale. Not more than 5.0 per cent, determined on 1.000 g by
Inject 100 pL of reference solution (d). When the drying in an oven at 60 °C over diphosphorus pentoxide R at a
chromatograms are recorded in the prescribed conditions, the pressure not exceeding 670 Pa for 3 h.
retention time for nitrite is 3.3 to 4.0 min. The test is not __________________________________________________________ PhEur
valid unless:
— the number of theoretical plates calculated for the nitrite
peak is at least 7000 per metre per column (dalteparin
sodium will block the binding sites of the stationary ★ ★
phase, which will cause shorter retention times and lower Danaparoid Sodium ★ ★
★. ★
separation efficiency for the analyte; the initial (Ph. Eur. monograph 2090) *★*
performance of the column may be partially restored
using a 58 g/L solution of sodium chloride R at a flow rate Chondroitin sulfate and
Heparan sulfate family
derm atari sulfate family
of 1.0 ml ymin for 1 h; after regeneration the column is
rinsed with 200 mL to 400 mL of water R );
— the symmetry factor for the nitrite peak is less than 3;
— the relative standard deviation of the peak area for nitrite
obtained from 6 injections is less than 3.0 per cent.
Inject 100 pL each of reference solutions (c) and (e).
The test is not valid unless:
— the correlation factor for a linear relationship between HO
concentration and response for reference solutions (c), (d) HO
and (e) is at least 0.995; — n
— the signal-to-noise ratio for reference solution (c) is not
less than 5 (if the noise level is too high, electrode
ADi R1 R2 R3 ADiHS R1 R2 R3
recalibration is recommended);
-OS H H H -OS H Ac H
— a blank injection of water R does not give rise to spurious
-6S S03Na H H -6S S03Na Ac H
peaks. -4S H S 0 3Na H -NS H S 0 3Na H
Inject 100 pL of the test solution. Calculate the content of -US H H S 0 3Na -US H Ac S03Na
nitrite from the peak areas in the chromatogram obtained -(U,6)S SOsNa H S 0 3Na -(U,N)S H S03Na S 0 3Na
with reference solutions (c), (d) and (e). -<U.4)S H S 03Na S 0 3Na
-(6,N)S S03Na S03Na H
-(4t6)S S03Na S 0 3Na H
Boron -(U,N,6)S S03Na S 03Na S 0 3Na
-<U,4,6)S S03Na SOsNa S 0 3Na
Not more than 1 ppm, determined by inductively coupled
plasma atomic emission spectroscopy.
83513-48-8
Boron is determined by measurement of the emission from
an inductively coupled plasma (ICP) at a wavelength specific Action and use
to boron. The emission line at 249.733 nm is used. Use an Heparinoid; prevention of deep vein thrombosis.
appropriate apparatus, whose settings have been optimised as
directed by the manufacturer. PhEur.
add and sulfuric add. It has the characteristic property of D etermine by selective enzymatic degradation.
enhancing the inactivation of activated factor X (factor Xa) Test solutions Dry the substance to be examined at 60 °C over
by antithrombin. It has a negligible effect on the inactivation diphosphorus pentoxide R at a pressure of about 670 Pa for
rate of thrombin by antithrombin. 3 h. Dissolve 0.200 g of the dried substance in 10.0 mL of
Potency water R. Dilute this solution as necessary to obtain 3 test
11.0 to 17.0 anti-factor Xa units per milligram (dried solutions containing 20 mg/mL, 10 mg/mL and 5 mg/mL of
substance). the dried substance to be examined in water R.
PRODUCTION Chondroitin sulfate reference solutions Dry chondroitin sulfate
The animals from which danaparoid sodium is derived must sodium CRS over diphosphorus pentoxide R at room
fulfil the requirements for the health of animals suitable for temperature at a pressure of about 670 Pa for 16 h. Prepare
human consumption. It is prepared using a process that solutions containing 1 mg/mL, 2 mg/mL and 3 mg/mL of
ensures that the relative proportion of active sulfated dried chondroitin sulfate sodium CRS in water R.
glycosaminoglycans is consistent. It is produced by methods Dermatan sulfate reference solutions Dry dermatan sulfate CRS
of manufacturing designed to minimise or eliminate over diphosphorus pentoxide R at room temperature at a
endotoxins and hypotensive substances. pressure of about 670 Pa for 16 h. Prepare solutions
CHARACTERS containing 1 mg/mL, 2 mg/mL and 3 mg/mL of dried
dermatan sulfate CRS in water R.
Appearance
White or almost white, hygroscopic powder. Chondroidnase A B C solution Dissolve chondroitinase A B C R in
tris-sodium acetate-sodium chloride buffer solution p H 8.0 R to
Solubility
Freely soluble in water. obtain an activity of 0.5-1.0 units per millilitre.
Chondroitinase A C solution Dissolve chondroitinase A C R in
IDENTIFICATION
tris-sodium acetate-sodium chloride buffer solution p H 7.4 R to
A. The ratio of anti-factor Xa activity to anti-factor Ha obtain an activity of 1.0-2.0 units per millilitre.
activity, determined as described under Assay and Tests
Procedure:
respectively, is not less than 22.
— Degradation with chondroidnase ABC: label 2 sets of
B. Molecular mass distribution (see Tests): the mass-average 10 tubes in triplicate: T l, T2 and T3 for the test
relative molecular mass ranges between 4000 and 7000. solutions; SD1, SD2 and SD3 for the dermatan sulfate
TESTS reference solutions; SCI, SC2 and SC3 for the
pH (2.2.3) chondroitin sulfate reference solutions; and B for the
5.5 to 7.0. blank (water R). To each tube add 1.25 mL of tris-sodium
Dissolve 0.5 g of the dried substance to be examined in acetate buffer solution p H 8.0 R and 150 pL of the test
carbon dioxide-free water R and dilute to 50 mL with the same solutions, dermatan sulfate reference solutions,
solvent. chondroitin sulfate reference solutions or water R.
A nti-factor Ila activity To each tube in 1 set of tubes add 75 jiL of
Maximum 0.5 units per milligram (dried substance). chondroitinase ABC solution. To determine the blank
response level, add 75 (iL of tris-sodium acetate-sodium
Test solutions Prepare 2 independent series of dilutions in
chloride buffer solution p H 8.0 R to each tube in the odaer
geometric progression of the substance to be examined in set of tubes. Mix the contents of the tubes using a vortex
phosphate buffer solution p H 6.5 R and in the concentration
mixer, cover with appropriate stoppers and incubate at
range of 0.0005 to 0.005 units of anti-factor Ha activity per 37 °C for at least 24 h.
millilitre.
— Degradation with chondroitinase AC: label 7 tubes in
Reference solutions Prepare 2 independent series of dilutions in triplicate: T l, T2 and T3 for the test solutions; SCI,
geometric progression of danaparoid sodium CRS in phosphate SC2 and SC3 for the chondroitin sulfate reference
buffer solution p H 6.5 R and in the concentration range of solutions; and B for the blank (water R). To each tube
0.0005 to 0.005 units of anti-factor Ha activity per millilitre. add 1.25 mL of tris-sodium acetate buffer solution p H 7.4 R
Transfer 50 jiL of each solution into the wells of a 96-well and 150 ^L of the test solutions,
microtitre plate. To each well add 50 jiL of antithrombin III chondroitin sulfate reference solutions or water R.
solution R3 and 50 |iL of human thrombin solution R l. Shake Add 75 jiL of chondroitinase AC solution to each tube.
the microtitre plate but do not allow bubbles to form. Mix the contents of the tubes using a vortex mixer, cover
Incubate for 75 min. To each well add 50 jiL of chromogenic with appropriate stoppers and incubate at 37 °C for at
substrate R4. Shake the microtitre plate. Measure the least 24 h. After the incubation period mix the contents
absorbances at 405 nm (2.2.25) using a suitable reading of the tubes using a vortex mixer and dilute to 12 times
device, exactly 4 min after the addition of the chromogenic with water R. Measure the absorbances (2.2.25) of the
substrate. The reaction may be stopped using 75 jiL of a diluted solutions at 234 nm against water R using a
20 per cent V/V solution of glacial acetic acid R. Determine suitable spectrophotometer.
the blank amidolytic activity in a similar manner, using Calculation Calculate the mean blank absorbance of each
phosphate buffer solution p H 6.5 R as the blank solution reference solution, i.e. the mean of die absorbances of the
(minimum 10 blanks per microtitre plate). Calculate the reference solutions to which no chondroitinase ABC has been
activity of the substance to be examined in units of anti added. Subtract the mean blank absorbance value from the
factor Ila activity per milligram using a suitable statistical individual absorbance of each reference solution. Calculate
method, for example the parallel-line assay. linear regression curves for the 2 chondroitin sulfate reference
Chondroitin sulfate and derm atan sulfate and the dermatan sulfate reference by plotting the blank-
Chondroitin sulfate: maximum 8.5 per cent (dried corrected absorbances against the concentrations.
substance); dermatan sulfate: 8.0 per cent to 16.0 per cent
(dried substance).
2016 Danaparoid Sodium 1-681
Calculate the average percentage content of dermatan sulfate determine the molecular mass distribution of the sample.
in the test solutions of all tested concentrations using the A calibration table is supplied with danaparoid sodium CRS.
following expression: Limits:
— chains with a relative molecular mass less than 2 0 0 0 .
a a (-^3 — A i — 1\) x B i T T maximum 13 per cent;
A2- Ai - ------------ =---- ------------h - h
— chains with a relative molecular mass less than 4000.
------------------ B % C ----------------- Xl0° maximum 39 per cent;
— chains with a relative molecular mass between 4000 and
Ai = blank absorbance of the test solution; 8000. minimum 50 per cent;
A2 = absorbance of the test solution with chondroitinase — chains with a relative molecular mass higher than 8000:
ABC; maximum 19 per cent;
A3 = absorbance of die test solution with chondroitinase — chains with a relative molecular mass higher than 1 0 000 '.
AC; maximum 11 per cent.
Bi = gradient of the curve obtained with the chondroitin Nitrogen (2.5.9)
sulfate reference solutions with chondroitinase AC; 2.4 per cent to 3.0 per cent (dried substance).
B2 = gradient of the curve obtained with the chondroitin Nucleic adds
sulfate reference solutions with chondroitinase ABC; Maximum 0.5 per cent (dried substance).
B3 = gradient of the curve obtained with the dermatan
Test solution Weigh about 50 mg of the dried substance to be
sulfate reference solutions with chondroitinase ABC;
C = concentration of the test solution, in milligrams per examined into a centrifuge tube and dissolve in 200 pL of
water R.
millilitre;
Ix = y-intercept of die curve obtained with the Reference solution Dissolve about 50 mg of ribonucleic
chondroitin sulfate reference solutions with acid CRS in 5 mL of 0.1 M sodium hydroxide and dilute to
chondroitinase AC; 20.0 mL with water R Transfer 200 pL of the solution into a
J2 = y-intercept of the curve obtained with the centrifuge tube.
chondroitin sulfate reference solutions with Add 4.0 mL of a 50 g/L solution of trichloroacetic acid R to
chondroitinase ABC; each tube and mix. Place all tubes in boiling water for
/3 = y-intercept of the curve obtained with the dermatan 30 min. Allow to cool to room temperature. Add again
sulfate reference solutions with chondroitinase ABC. 4.0 mL of a 50 g/L solution of trichloroacetic acid R to each
Calculate the average percentage content of chondroitin tube and mix. If any of the test solutions is not clear,
sulfate in the test solutions for all tested concentrations using sonicate all the tubes in an ultrasonic bath for 10 m in and
the following expression: centrifuge at 1500 g for 15 min. Dilute 1.0 mL of the clear
supernatant to 4.0 mL with water R. Measure the
(A 3 - A x - I x ) x 100 absorbances of the diluted reference and test solutions at
265 nm (2.2.25) against a blank solution prepared in the '
BxxC
same manner, and calculate the percentage nucleic acid 1
content of the sample.
M olecular mass distribution
Size-exclusion chromatography (2.2.30). Total protein (2.5.33, Method 2)
M axim um 0.5 per cent.
Test solution Dissolve 10 mg of the substance to be examined
in 2 mL of the mobile phase. Dissolve the substance to be examined in water R. Use bovine
albumin R as the reference substance.
Reference solution Dissolve 10 mg of danaparoid sodium CRS in
2 mL of the mobile phase. Sodium
Column:
9.0 per cent to 11.0 per cent (dried substance).
— size: I = 0.60 m, 0 = 7.5 mm; Atomic absorption spectrometry (2.2.23, Method I).
— stationary phase: hydropMHc silica gd for chromatography R Test solution Dissolve 0.125 g of the substance to be
(10 pm) with a fractionation range for proteins with a examined in 100.0 mL of a 1.27 mg/mL solution of caesium
relative molecular mass of approximately 5000-100 000; chloride R in 0.1 M hydrochloric acid.
— temperature: 30 °C. Reference solutions Prepare reference solutions containing
Mobile phase 28.4 g/L solution of anhydrous sodium sulfate R 50 ppm, 100 ppm and 150 ppm of Na by diluting sodium
adjusted to pH 5.0 with dilute sulfuric acid R standard solution (1000 ppm Na) R with a 1.27 mg/mL
Flow rate 0.9 ml 7min ± 2 per cent solution of caesium chloride R in 0.1 M hydrochloric add.
Detection Spectrophotometer at 210 nm. Source Sodium hollow-cathode lamp.
Run time For a period of time ensuring complete elution of Atomisation device Air-acetylene flame.
sample and solvent peaks (about 40 min). Loss on drying (2.2.32)
System suitability Inject the reference solution twice. Maximum 5.0 per cent, determined on 0.500 g by drying in
The difference between the retention times corresponding to an oven at 60 °C over diphosphorus pentoxide R at a pressure
the maxima of the peaks is not more than 5 s. of 670 Pa for 3 h.
Calibration Calibration is achieved by taking the relevant part Bacterial endotoxins (2.6.14)
of the chromatogram obtained with the reference solution, Less than 0.02 IU per unit of anti-factor Xa activity, if
i.e. excluding the sharp peak at the end of the intended for use in the manufacture of parenteral
chromatogram, and matching the chromatogram obtained preparations without a further appropriate procedure for the
with the test solution with the calibration table obtained with removal of bacterial endotoxins.
the reference solution. From the calibration curve obtained,
1-682 Dantrolene Sodium 2016
ASSAY CHARACTERISTICS
The anticoagulant activity of danaparoid sodium is A yellowish-orange to orange crystalline powder.
determined in vitro by an assay which determines its ability to Very slightly soluble in water, slightly soluble in ethanol
accelerate the inhibition of factor Xa by antithrombin HI (96%); sparingly soluble in methanol’, practically insoluble in
(anti-factor Xa assay). acetone.
Test solutions Prepare 2 independent series of dilutions in
IDENTIFICATION
geometric progression of the substance to be examined in
A. The infrared absorption spectrum, Appendix II A, is
tris(hydroxymethyl)aminomethane ED TA buffer solution
concordant with the reference spectrum of dantrolene sodium
p H 8.4 R and in the concentration range of 0.1 to 0.32 units
CRS 422).
of anti-factor Xa activity per millilitre.
B. In the Assay, the chromatogram obtained with solution
Reference solutions Prepare 2 independent series of dilutions in
(1) shows a peak with the same retention time as the
geometric progression of danaparoid sodium CRS in
principal peak in the chromatogram obtained with
tris(hydroxyrnethyl)aminomethane ED TA buffer solution
solution (2).
pH 8.4 R and in the concentration range of 0.08 to
0.35 units of anti-factor Xa activity per millilitre. C. To 0.1 g of the substance being examined add 20 mL of
water and 2 drops of acetic acid, shake well and filter.
Transfer 40 |iL of each solution into the wells of a 96-well
The filtrate yields the reactions characteristic of sodium salts,
microtitre plate. Add 40 jiL of antithrombin III solution R4 to
Appendix VI.
each well and shake the microtitre plate but do not allow
bubbles to form. Add 40 jiL of bovine factor X a solution R1 to TESTS
each well. Exactly 2 min after the addition of the factor Xa Alkalinity
solution, add 80 pL of chromogenic substrate R5. Measure the Shake 0.7 g in 10 mL of water for 5 minutes and centrifuge.
absorbance at 405 nm (2.2.25) using a suitable reading To 5 mL of die supernatant add 45 mL of water and 3 drops
device, exactly 4 min after the addition of the factor Xa of phenolphthalein solution R1 and 0.1 m l. of 0.1m hydrochloric
solution. The reaction may be stopped using 75 pL of a acid VS. A red colour is not produced.
20 per cent VIV solution of glacial acetic acid R. Determine Related substances
the blank amidolytic activity in the same manner, using Carry out the method for liquid chromatography,
tris(hydroxymethyl)aminomethane ED TA buffer solution Appendix IQ D, using the following solutions.
pH 8.4 R as the blank (minimum 8 blanks per microtitre
(1) Dissolve 50 mg of the substance being examined in
plate). Calculate the potency of the substance to be
20 mL of tetrahydrcfuran and 2 mL of glacial acetic acid and
examined in units of anti-factor Xa activity per milligram
dilute with sufficient absolute ethanol to produce 100 mL.
using a suitable statistical method, for example the parallel-
line assay. (2) Dilute 1 mL of solution (1) to 100 mL with absolute
ethanol.
STORAGE
(3) Dissolve 5 mg of dantrolene sodium BPCRS and 0.1 g of
In an airtight container. If the substance is sterile, store in a theophylline BPCRS in 20 mL of tetrahydrcfuran and 2 mL of
sterile, airtight, tamper-proof container. glacial acetic acid and dilute with sufficient absolute ethanol to
LABELLING produce 100 mT„ Further dilute 10 mL of this solution to
The label states the number of units of anti-factor Xa activity 100 m l. with absolute ethanol
per milligram. C H R O M A T O G R A P H IC C O N D IT IO N S
___________________________________________________________PhEtr (a) Use a stainless steel column (15 cm x 4.6 mm) packed
with silica gelfor chromatography (5 Jim) (Zorbax Sil is
suitable).
(b) Use isocratic elution and the mobile phase described
Dantrolene Sodium below.
(c) Adjust the flow rate of the mobile phase so that the
retention time of the peak corresponding to Dantrolene
Sodium is about 8 minutes.
(d) Use a column temperature of 30°.
NNa , 31/2 HzO
(e) Use a detection wavelength of 300 nm.
(f) Inject 10 |iL of each-solution.
(g) For solution (1) allow the chromatography to proceed for
at least twice the retention time of the principal peak.
C14H 9N4Na05 ,31/ 2H20 399.3 24868-20-0
MOBILE PHASE
Action and use 9 volumes of absolute ethanol, 10 volumes of glacial acetic acid
Skeletal muscle relaxant and 90 volumes of hexane.
Preparation SY ST E M S U IT A B IL IT Y
Dantrolene Oral Suspension The test is not valid unless, in the chromatogram obtained
with solution (3), the resolutionfactor between the peaks
DEFINITION
corresponding to theophylline and dantrolene is at least 6.
Dantrolene Sodium is l-(5-p-
nitrophenylfurfurylideneamino)hydantoin sodium. It contains L IM IT S
not less than 98.0% and not more than 102.0% of In the chromatogram obtained with solution (1):
Ci4H 9N 4Na 0 5 , calculated with reference to the anhydrous
substance.
2016 Dantron 1-683
the total area of all the secondary peaks is not greater than the CHARACTERISTICS
area of the principal peak in the chromatogram obtained with An orange, crystalline powder.
solution (2) (1%). Practically insoluble in water, slightly soluble in ether, very
Heavy metals slightly soluble in ethanol (96%). It dissolves in solutions of
1.0 g complies with limit test C for heavy metals, alkali hydroxides.
Appendix VII. Use 2 m l. of lead standard solution IDENTIFICATION
(10 ppm Pb) to prepare the standard (20 ppm).
A. The infrared absorption spectrum, Appendix II A, is
Water concordant with the reference spectrum of dantron (RS 083).
14.5 to 17.0% w/w, Appendix IX C. Use 0.2 g B. The light absorption, Appendix IIB , in the range 230 to
ASSAY 350 nm of a 0.001% w/v solution in dichbromethane exhibits
Carry out the method for liquid chromatography, maxima at 255 nm and 285 nm and a less well-defined
Appendix HI D, using the following solutions. maximum at 275 nm. The absorbance at the maximum at
(1) Dissolve 60 mg of the substance being examined in 255 nm is about 0.82 and at the maximum at 285 nm is
50 mL of dimethylformamide and dilute 1 volume of the about 0.48, each calculated with reference to the dried
resulting solution to 100 volumes with the mobile phase. substance.
(2) Dilute 1 volume of a 0.12% w/v solution of dantrolene C. Dissolve 5 mg in 5 m l. of 1m sodium hydroxide. A clear
sodium BPCRS in dimethylformamide to 100 volumes with the
red solution is produced immediately.
mobile phase. TESTS
C H R O M A T O G R A P H IC C O N D IT IO N S Mercury
(a) Use a stainless steel column (15 cm x 4.6 mm) packed To 0.50 g in a Kjeldahl flask add 2.5 mL of nitric acid and
with spherical particles of silica, 5 pm in diameter, the allow to stand until the initial vigorous reaction has subsided.
surface of which has been modified with chemically-bonded Add 2.5 mL of sulfuric acid and heat until dense white fumes
nitrile groups (Spherisorb CN is suitable). are evolved. Cool, add 2.5 mL of nitric add and heat until
fumes are again evolved. Repeat the procedure with a further
(b) Use isocratic elution and the mobile phase described 2.5 mL of nitric acid, cool, add 50 mL of water, boil the
below. solution until the volume has been reduced to about 25 mL
(c) Use a flow rate of 1 mL per minute. and cool. Transfer to a separating funnel using water, dilute
(d) Use an ambient column temperature. to about 50 mL with water and add 50 mL of 0.5m sulfuric
(e) Use a detection wavelength of 262 nm. add. Add 100 mL of water, 2 g of hydroxylamine
hydrochloride, 1 m l. of 0.05m disodium edetate, 1 ml. of glacial
(f) Inject 20 pL of each solution.
acetic add and 5 mL of dichbromethane, shake, allow to
MOBILE PHASE separate and discard the dichloromethane layer. Titrate the
15 volumes of acetonitrUe and 85 volumes of a phosphate aqueous layer with a 0.0008% w/v solution of dithizone in
buffer pH 6.8 prepared by dissolving 11.88 g of disodium dichbromethane, shaking vigorously after each addition,
hydrogen orthophosphate and 9.08 g of potassium dihydrogen allowing the layers to separate and discarding the
orthophosphate in 1000 m l. of water. dichloromethane layer, until the dichloromethane layer
D E T E R M IN A T IO N O F C O N T E N T
remains green. Repeat the operation using a solution
prepared by diluting 1 mL of mercury standard solution
Calculate the content of C 14H 9N 4Na 0 5 in the substance
(5 ppm Hg) to 100 mL with 0.5m sulfuric acid and beginning
being examined using the declared content of Ci4H9N4N a0 5
at the words ‘Add 100 mL of water ... ’. The volume of the
in dantrolene sodium BPCRS.
dithizone solution required by the substance being examined
does not exceed that required by the mercury standard
solution.
Related substances
Dantron Carry out the method for liquid chromatography,
Appendix EH D, using the following solutions.
OH O OH (1) Dissolve 50 mg of the substance being examined in
20 m l. of tetrahydrofuran and dilute to 100 mL with the
mobile phase.
(2) Dilute 1 volume of solution (1) to 50 volumes with the
O mobile phase.
(3) Dissolve 50 mg of dantron impurity standard BPCRS in
20 ml. of tetrahydrofuran and dilute to 100 mL with the
Ci4H80 4 240.2 117-10-2
mobile phase.
Action and use C H R O M A T O G R A P H IC C O N D IT IO N S
Anthraquinone stimulant laxative. (a) Use a stainless steel column (25 cm x 4.6 mm) packed
Preparation with octadecylsUyl silica gelfor chromatography (5 jim)
Co-danthrusate Capsules (Nucleosil C18 is suitable).
(b) Use an isocratic system using the mobile phase described
DEFINITION below.
Dantron is mainly 1,8-dihydroxyanthraquinone. It contains
(c) Use a flow rate of 1 mL per minute.
not less than 98.0% and not more than 102.0% of total
phenols, calculated as C 14H 804 and with reference to the (d) Use an ambient column temperature.
dried substance. (e) Use a detection wavelength of 254 nm.
1-684 Dapsone 2016
IMPURITIES IDENTIFICATION
A. The light absorption, Appendix IIB , in the range 230 to
0 OH
350 nm of a 0.05% w/v solution in 0.05m sulfuric acid
exhibits two maxima, at 262 nm and 270 nm. The absorbance
at the m a x i m u m at 262 nm is about 0.69 and at the
m a x i m u m at 270 nm is about 0.51.
Debrisoquine Sulphate C H R O M A T O G R A P H IC C O N D IT IO N S
Sulfated ash
Not more than 0.1%, Appendix IX A.
Demeclocycline Hydrochloride *****
ASSAY (Ph. Eur. monograph 0176) *
Carry out Method I for rum-aqueous titration,
Appendix VIII A, using 1 g and determining the end point
potentiometrically. Each mL of 0 .1 m perchloric acid FS is
equivalent to 44.85 mg of (CioHi3N 3) 2jH 2S0 4 .
STORAGE
Debrisoquine Sulfate should be protected from light.
PhEtr______________________ DEFINITION
DEFINITION (4S,4aSj5aS,6S,12aS)-7-Chloro-4-(dimethylamino)-
Mixture consisting of decyl esters of fatty acids, mainly oleic 3,6,10,12,12a-pentahydroxy-l,l l-dioxo-l,4,4a,5,5a,6,l 1,12a-
(as-9-octadecenoic) acid. octahydrotetracene-2-carboxamide hydrochloride.
A suitable antioxidant may be added. Substance produced by certain strains of Streptomyces
aureofaciens.
CHARACTERS
Content
Appearance
89.5 per cent to 102.0 per cent (anhydrous substance).
Clear, pale yellow or colourless liquid.
Solubility CHARACTERS
Practically insoluble in water, misdble with ethanol Appearance
(96 per cent), with methylene chloride and with light Yellow powder.
petroleum (bp: 40-60 °C). Solubility
Soluble or sparingly soluble in water, slighdy soluble in
IDENTIFICATION
alcohol, very slightly soluble in acetone. It dissolves in
A. Relative density (see Tests).
solutions of alkali hydroxides and carbonates.
B. Saponification value (see Tests).
IDENTIFICATION
C. Oleic add (see Tests).
A. Thin-layer chromatography (2.2.27).
TESTS Test solution Dissolve 5 mg of the substance to be examined
Relative density (2.2.5) in methanol R and dilute to 10 mL with the same solvent.
0.860 to 0.870. Reference solution (a) Dissolve 5 mg of demeclocycline
A dd value (2.5.1) hydrochloride CRS in methanol R and dilute to 10 mL with the
Maximum 1.0, determined on 10.0 g. same solvent.
Iodine value (2.5.4, Method A) Reference solution (b) Dissolve 5 mg of demeclocycline
55 to 70. hydrochloride CRS, 5 mg of chlortetracycHne hydrochloride R and
Peroxide value (2.5.5, Method A) 5 mg of tetracycline hydrochloride R in methanol R and dilute to
Maximum 10.0. 10 mL with the same solvent.
Saponification value (2.5.6) Plate T L C octadecylsilyl silica gel F 254 plate R.
130 to 140, determined on 2.0 g. Mobile phase Mix 20 volumes of acetonitrUe R, 20 volumes of
methanol R and 60 volumes of a 63 g/L solution of oxalic
Oleic a d d (2.4.22, Method A)
acid R previously adjusted to pH 2 with concentrated
Minimum 60.0 per cent in the fatty add. fraction of the
ammonia R.
substance.
Application 1 |oL.
Water (2.5.12)
Maximum 1.0 per cent, determined on 1.00 g. Development Over 3/4 of the plate.
Drying In air.
Total ash (2.4.16)
Maximum 0.1 per cent, determined on 2.0 g. Detection Examine in ultraviolet light at 254 nm.
System suitability The chromatogram obtained with reference
STORAGE
solution (b) shows 3 clearly separated spots.
Protected from light.
Results The prindpal spot in the chromatogram obtained with
______________________________________ ___________________ PhEur
the test solution is similar in position and size to the prindpal
spot in the chromatogram obtained with reference
solution (a).
1-688 Demeclocycline Hydrochloride 2016
Deptropine Citrate ★ ★
★ ★
(Ph. Eur. monograph 1308) * * * * *
HO H H3C —N H
CH3
HO C02H
C. (42?j4a5j5a5j65j 12a5)-4-(dimethylamino)-3j6j 10,12,12a- > H02C ^ y ^ C 0 2H
pentahydroxy-1,1 l-dioxo-l,4,4a,5,5a,6,l 1,12a-
octahydrotetxacene-2-carboxamide
(4-epidemethyltetxacycline),
DEFINITION
1,1 '-(Decane-1,10-diyl) bis (4-amino-2-methylquinolinium)
dichloride (dried substance).
Content
95.0 per cent to 101.0 per cent.
A. (1i?,3r,55)-8-methyl-8-azabicyclo[3.2.l]octan-3-ol CHARACTERS
(tropine), Appearance
White or yellowish-white powder, hygroscopic.
Solubility
Slightly soluble in water and in ethanol (96 per cent).
IDENTIFICATION
First identification: Bs E.
2016 Dequalinium Chloride 1-691
Second identification A , C, D, E. curve separating this peak from the peak due to
A. Ultraviolet and visible absorption spectrophotometry dequalinium chloride. If necessary, adjust the
(2.2.25). concentration of methanol in the mobile phase.
Test solution Dissolve
about 10 mg in water R and dilute to Limits:
100 mL with the same solvent. Dilute 10 mL of the solution — impurity A: not more than 0.5 times the area of the
to 100 mL with water R. principal peak in the chromatogram obtained with
Spectral range 230-350 nm.
reference solution (b) (1 per cent);
— total of impurities other than A: not more than 5 times the
Absorption maxima At 240 nm and 326 nm.
area of the principal peak in the chromatogram obtained
Shoulder At 336 nm. with reference solution (b) (10 per cent);
Absorbance ratios: — disregard limit: 0.025 times the area of the principal peak
— A2WA326 = 1-56 to 1.80; in the chromatogram obtained with reference solution (b)
— A32<j/A33g = 1.12 to 1.30. (0.05 per cent).
B. Infrared absorption spectrophotometry (2.2.24). Readily carbonisable substances
Spectral range 600-2000 cm-1. Dissolve 20 mg in 2 mL of sulfuric add R. After 5 min the
Comparison dequalinium chloride CRS. solution is not more intensely coloured than reference
solution BY4 (2.2.2, Method I).
C. To 5 ml, of solution S (see Tests) add 5 mL of potassium
ferricyanide solution R. A yellow precipitate is formed. Loss on drying (2.2.32)
D. To 10 mL of solution S add 1 mL of dilute nitric add R. Maximum 7.0 per cent, determined on 1.000 g by drying at
A white precipitate is formed. Filter and reserve the filtrate 105 °C at a pressure not exceeding 0.7 kPa.
for identification test E. Sulfated ash (2.414)
E. The filtrate from identification test D gives reaction (a) of Maximum 0.1 per cent, determined on 1.0 g.
chlorides (2.3.1). ASSAY
TESTS In order to avoid overheating in the reaction medium, mix
Solution S thoroughly throughout and stop the titration immediately after the
Dissolve 0.2 g in 90 mL of carbon dioxide-five water R, end-point has been reached
heating if necessary, and dilute to 100 mL with the same Dissolve 0.200 g in 5 ml, of anhydrous formic add R and add
solvent. 50 mL of acetic anhydride R. Titrate with 0.1 M perchloric
add, determining the end-point potentiometrically (2.2.20).
Appearance of solution
Solution S is clear (2.2.1) and colourless (2.2.2, Method II). 1 mL of 0.1 M perchloric add is equivalent to 26.38 mg of
C 30H 40CI2N 4.
Acidity or alkalinity
To 5 mL of solution S add 0.1 mL of bromothymol blue STORAGE
solution R l. Not more than 0.2 mL of 0.01 M hydrochloric In an airtight container.
acid or 0.01 M sodium hydroxide is required to change the
IMPURITIES
colour of the indicator. Specified impurities A.
Related substances Other detectable impurities (the following substances would, if
Liquid chromatography (2.2.29). present at a sufficient level, be detected by one or other of
Test solution Dissolve 10.0 mg of the substance to be the tests in the monograph. They are limited by the general
examined in the mobile phase and dihate to 10.0 mL with acceptance criterion for other/unspecified impurities and/or
the mobile phase. by the general monograph Substances for pharmaceutical use
Reference solution (a) Dissolve 10.0 mg of dequalinium chloride (2034). It is therefore not necessary to identify these
for performance test CRS in the mobile phase and dilute to impurities for demonstration of compliance. See also 5.10.
10.0 mL with the mobile phase. Control of impurities in substancesfor pharmaceutical use): B, C.
Reference solution (b) Dissolve 10.0 mg of dequalinium
chloride CRS in the mobile phase and dilute to 10.0 mL with
the mobile phase. Dilute 1.0 mL of the solution to 50.0 mL
with the mobile phase.
Column:
— size. I = 0.25 m, 0 = 4.6 mm; ch 3
— stationary phase: end-capped octadecylsûyl silica gel for
chromatography R.
A. 2-methylquinolin-4-amine,
Mobile phase Dissolve 2 g of sodium hexanesidfonate R in
300 mL of water R~, adjust to pH 4.0 with acetic add R and
add 700 mL of methanol R.
Flow rate 1.5 mL/min.
Detection Spectrophotometer at 240 nm.
Injection 10 |iL.
Run time 5 times the retention time of dequalinium chloride.
System suitability: reference solution (a):
B. 4-amino-1- [10- [(2-methylquinolin-4-yl) amino] decyl] -2-
— peak-to-vaüey ratio: minimum 2.0, where H p = height
methylquinolinium chloride,
above the baseline of the peak due to impurity B and
H v = height above the baseline of the lowest point of the
1-692 3-0-Desacyl-4’-Monophosphoryl lip id A 2016
— tetraacyl: 15 per cent to 35 per cent; liquid bulk is within the limits approved for the particular
— pentaacyl: 35 per cent to 60 per cent; product.
— hexaacyl: 20 per cent to 40 per cent; Sterility (2.6.1)
— heptaacyl: less than 0.5 per cent. It complies with the test, carried out losing 10 mL for each
Triethylamine medium.
4.2 to 5.8 per cent m/m, determined by a suitable method, Congener distribution
for example gas chromatography (2.2.28). The relative amount of tetraacyl, pentaacyl, hexaacyl and
Water (2.5.12) heptaacyl congener groups are determined by a suitable
Maximum 6.7 per cent mlm. method, for example reversed-phase HPLC analysis (2.2.29).
Free fatty acids The relative amount of each congener group in the 3-0-
Maximum 2.6 per cent mlm, determined by a suitable desacyl-4'-monophosphoryl lipid A liquid bulk is:
method, for example reversed-phase HPLC analysis (2.2.29). — tetraacyl: 15 per cent to 35 per cent;
2-Keto-3-deoxyoctonate — pentaacyl: 35 per cent to 60 per cent;
Less than 0.5 per cent m/m, determined by a suitable — hexaacyl: 20 per cent to 40 per cent;
method. For example, a colorimetric method may be used — heptaacyl: less than 0.5 per cent.
where 2-keto-3-deoxyoctonate is released by hydrolysis (0.2 ASSAY
N H2S 0 4 at 100 °C for 30 min), oxidised by periodic acid, The 3-0-desacyl-4'-monophosphoryl lipid A content is
and reacted with sodium arsenite to yield P-formylpyruvic determined by a suitable method, for example gas
acid, which subsequently is coupled to thiobarbituric acid to chromatographic quantification (2.2.28) of trifluoroacetic
give a red coloured chromophore with absorption maximum anhydride derivatised fatty add methyl esters of the 3-0-
at 550 nm. The amount of 2-keto-3-deoxyoctonate is desacyl-4'-monophosphoryl lipid A fatty adds dodecanoic
interpolated from a calibration curve. add (C l2:0), tetradecanoic add (C l4:0), 3-hydroxy
Identity tetradecanoic add (3-OH-Cl4:0) and hexadecanoic add
The test for congener distribution also serves to identify the (C16:0) obtained by hydrolysis of 3-0-desacyl-4'-
product monophosphoryl lipid A in an aqueous/methanol (50:50 VIV)
solution, containing 5 per cent of sodium hydroxide. To the
Microbial contamination
test sample, a reference sample and the dilutions of the
TAMC: acceptance criterion 101 CFU/10 mg (2.6.12).
calibration curve, pentadecanoic add (C l5:0) is added as an
Pyrogens (2.6.8) internal standard. The temperature gradient applied must
The triethylamine salt of 3-0-desacyl-4 '-monophosphoryl allow the separation of the fatty add methyl esters in about
lipid A complies with the test for pyrogens. Inject into each 40 min.
rabbit per kilogram of body mass 3 mL of a solution
The siim of the ratios between the area for each individual
containing 2.5 ng of 3-0-desacyl-4'-monophosphoryl lipid A.
fatty add methyl ester (C12:0, C14:0, 3-OH-Cl4:0 and
3-0-DESACYL-4' -MONOPHOSPHORYL UPID A LIQUID C l6:0) and the area of the internal standard (ratio = area
BULK Qc / area C15:0) is calculated The 3-0-desacyl-4'-
The triethylamine salt of 3-0-desacyl-4'-monophosphoryl monophosphoryl lipid A quantity corresponding to the sum
lipid A is dispersed in a liquid suitable for the subsequent ratio value on the calibration curve, established with the
processing steps at a defined target concentration. If the salt dilutions of the 3-0-desacyl-4'-monophosphoryl lipid A
is not soluble in water a microfluidisation step is necessary to standard, is reported.
prepare a stable aqueous suspension. The content of 3-0-desacyl-4'-monophosphoryl lipid A is
The liquid bulk is sterilised by filtration through a bacteria- not less than 80 per cent and not greater than 120 per cent
retentive filter. of the estimated content.
Only a 3-0-desacyl-4 '-monophosphoryl lipid A liquid bulk PhEur
that complies with the requirements given below under
Identification, Tests and Assay and that is within the limits
approved for the particular product may be used for the
preparation of 3-0-desacyl-4'-monophosphoryl lipid A in the
final lots. Desferrioxamine Mesilate ★ ★
★ ★
CHARACTERS (Deferoxamine Mesilate, Ph Eitr monograph 0896) *****
When dispersed in an aqueous solution: slightly turbid
suspension. OH
When dissolved in an organic solvent: a description of its
appearance is established and approved by the competent S03H
authority; the 3-0-desacyl-4'-monophosphoryl lipid A liquid
ch 3
bulk complies with this description.
IDENTIFICATION
Congener distribution (see Tests).
TESTS C26H52N5O11S 657 138-14-7
Particle size
Where applicable, the particle size in the microfluidised Action and use
liquid bulk is determined by a suitable method, for example Chelating agent (iron).
dynamic light scattering. The particle size for each batch of Preparation
Desferrioxamine Injection
1-694 Desfemoxamine Mesilate 2016
— impurities E, H: for each impurity, not m ore than the Reference solutions T o each of 1.0 m T, 2.0 mT, 3.0 mT.,
difference between the area of the corresponding peak in 4.0 m L and 5.0 m L o f antimony standard solution (100 ppm
the chromatogram obtained with reference solution (a) Sb) R add 20 m L of the solvent mixture and dilute to
and the area o f th e corresponding peak in the 100.0 m L with water R.
chromatogram obtained with the test solution Source Antimony hollow-cathode lam p using a transmission
(0.01 per cent V/V)’, band of 0.2 nm and a 75 per cent lamp current.
— impurity F: not m ore than the difference between the area Wavelength 217.6 nm.
of the corresponding peak in the chromatogram obtained
with reference solution (a) and the area o f the
Atomisation device Air-acetylene flame.
corresponding peak in the chromatogram obtained with N o n -v o latile m a tte r
the test solution (0.002 per cent V/V)', Maximum 100 mg/L.
— unspecified impurities: for each impurity, no t more than Evaporate 20.0 m L to dryness with the aid of a stream of
0.5 times the difference between the area o f the peak due nitrogen R. T h e residue weighs not more than 2.0 mg.
to impurity A in the chromatogram obtained with
STORA GE
reference solution (b) and the area o f the peak due to
In a glass botde fitted with a polyethylene-lined cap. Before
impurity A in the chromatogram obtained with the test
opening the botde, cool the contents to below 10 °C.
solution (0.005 per cent V/V)-,
— sum of impurities other than A, B, C, D, E, F, G and H: IM P U R IT IE S
not m ore than th e difference between the area of the peak Specified impurities A, B, C, D, E, F , G, H
due to impurity A in the chromatogram obtained with
reference solution (b) and the area of the peak due to F F
impurity A in th e chromatogram obtained with th e test
solution (0.01 p er cent V/V)-, F a C ^ 'O '^ 'C F j
— disregard Unde, the difference between the area of the peak
due to impurity A in the chromatogram obtained with A. 1,1 '-oxybis( 1,2,2,2-tetrafluoroethane),
reference solution (c) and the area o f the peak due to
impurity A in the chromatogram obtained with the test h a f
solution (0.002 per cent V/V). J. and enantiomer
f 3c o f
F lu o rid e s
M axim um 10 ppm .
B. (22?S)-2-chloro-2-(difluoromethoxy)-l,l,l-trifluoroethane
Potentiom etry (2.2.36, Method I).
(isofluorane),
Test solution T o 10.0 m L in a separating funnel, add 10 m L
o f a mixture o f 30.0 m L of dilute ammonia R2 and 70.0 m i. R R’
o f distilled water R. Shake for 1 min and collect the upper
layer. Repeat this extraction procedure twice, collecting the 0 ,^ 0 ,
upper layer each time. Adjust the combined upper layers to
p H 5.2 with dilute hydrochloric acid R. Add 5.0 m L o f fluoride C. R = H , R ' = F: dichlorofluoromethane,
standard solution (1 ppm F) R and dilute to 50.0 m L with D . R = Cl, R ' = F: trichlorofluoromethane,
distilled water R. T o 20.0 m L o f this solution add 20.0 m L of E. R = R ' = H : dichloromethane (methylene chloride),
totd-iomc-strength-adjustment buffer R and dilute to 50.0 m L
F. R = H , R ' = Cl: trichloromethane (chloroform),
w ith distilled water R.
Reference solutions T o each of 1.0 mL, 2.0 m L, 3.0 m L , ci ci
4.0 m L and 5.0 m L o f fluoride standard solution (10 ppm F) R
add 20.0 m L of total-ionic-strength-adjustment buffer R and
dilute to 50.0 m L w ith distilled water R. F F
3.0 m L of alkaline sodium picrate solution R and 5.0 m L of Reference solution (c) Dissolve 4 m g o f desloratadine for system
alcohol R prepared at the same time. suitability CRS (containing impurities A and B) in the mobile
Calculate the content o f C 47H 74019 from the absorbances phase and dilute to 5.0 m L with the mobile phase. Dilute
measured and th e concentrations of the solutions. 1.0 m L of the solution to 10.0 m L w ith the mobile phase.
STORAGE
Column:
— sizer. I = 0.25 m, 0 = 4.6 mm;
Store in an airtight, glass container, protected from light, at a
— stationary phase: end-capped octadecylsUyl silica gel for
tem perature below 10 °C.
chromatography R (4 |im);
__________________________________________________________ PhEur — temperature: 35 °C.
Mobile phase Dissolve 0.865 g of sodium dodecyl sulfate R in
water R, add 0.5 m L o f trifliwroacetic acid R and dilute to
1000 m L with water R; mix 57 volumes of this solution and
Desloratadine ***** 43 volumes *of acetonitrile R.
Flow rate 1.0 mL/min.
(Ph. Eur. monograph 2570) *
Detection Spectrophotometer at 280 run.
Injection 100 (iL of the test solution and reference
solutions (b) and (c).
Run time 2.5 times the retention time o f desloratadine.
Identification of impurities Use the chromatogram supplied
with desloratadine for system suitability CRS and the
chromatogram obtained with reference solution (c) to identify
the peaks due to impurities A and B.
C 19H i 9C1N2 310.8 100643-71-8 Relative retention W ith reference to desloratadine (retention
time = about 21 min): impurity A = about 0.8;
A ctio n a n d u se impurity B = about 0.9.
H istamine H l5 receptor .antagonist; antihistamine. System suitability: reference solution (c):
— resolution: minimum 2.0 between the peaks due to
PhEur__________________________________________________________
im purity B and desloratadine.
D E F IN IT IO N Calculation of percentage contents:
8-C hloro-l l-(piperidin-4-ylidene)-6,l l-dihydro-5//- — correction factors: multiply the peak areas o f the following
benzo [5,6] cyclohepta [ 1,2-6] pyridine. impurities by the corresponding correction factor:
C o n te n t impurity A = 1.6; impurity B = 1.6;
98.0 per cent to 102.0 per cent (anhydrous substance). — for each impurity, use the concentration of desloratadine
in reference solution (b).
CHARACTERS
Limits:
A p p e a ra n c e
— impurity B: maximum 0.3 per cent;
White or almost white powder.
— impurity A: maximum 0.2 per cent;
Solubility — unspecified impurities: for each impurity, maximum
Very slightly soluble o r pratically insoluble in water, freely 0.10 per cent;
soluble in ethanol (96 per cent), slightly soluble o r very — total: m aximum 0.4 per cent;
slightly soluble in heptane. — reporting threshold: 0.05 per cent.
It shows polymorphism (5.9). H eav y m e ta ls (2.4.8)
ID E N T IF IC A T IO N M aximum 20 ppm.
Infrared absorption spectrophotometry (2.2.24). Solvent methanol R.
Comparison desloratadine CRS. 0.250 g complies with test H . Prepare the reference solution
If the spectra obtained in the solid state show differences, using 0.5 m L o f lead standard solution (10 ppm Pb) R.
dissolve the substance to be examined and the reference W a te r (2.5.32)
substance separately in methyl isobutyl ketone R, evaporate to M aximum 0.5 per cent, determined on 0.250 g.
diyness and record new spectra using the residues. S u lfa te d a sh (2.4.14)
TESTS M aximum 0.2 per cent, determined on 0.5 g.
R e la te d su b s ta n c e s ASSAY
Liquid chrom atography (2.2.29).
Liquid chromatography (2.2.29) as described in the test for
Test solution Dissolve 20.0 mg o f the substance to be related substances with the following modifications.
examined in the mobile phase and dilute to 25.0 m L with
Injection T est solution and reference solution (a).
the mobile phase. D ilute 5.0 m L of the solution to 50.0 m L
System suitability: reference solution (a):
with the mobile phase.
— symmetry factor. 0.5 to 1.5 for the peak due to
Reference solution (a) Dissolve 20.0 mg of desloratadine CRS in desloratadine.
the mobile phase and dilute to 25.0 m L with the mobile
Calculate the percentage content of Q 9H 19CIN 2 taking into
phase. Dilute 5.0 m L o f the solution to 50.0 m L with the
account the assigned content o f desloratadine CRS.
mobile phase.
Reference solution (b) Dilute 1.0 m L o f the test solution to IMPURITIES
100.0 m L with th e mobile phase. Dilute 1.0 .m L o f this Specified impurities A, B.
solution to 10.0 m L with the mobile phase.
1-700 Desmopressin 2016
PhEur. 40 - 50 76 24
IM P U R IT IE S
Solubility
Practically insoluble in water, very soluble in methanol, freely
Other detectable impurities (the following substances would, if
soluble in anhydrous ethanol and in methylene chloride.
present at a sufficient level, be detected by one or other o f
the tests in the monograph. They are limited by the general IDENTIFICATION
acceptance criterion for other/unspecified impurities and/or A. Infrared absorption spectrophotom etry (2.2.24).
by the general monograph Substances for pharmaceutical use Comparison desogestrel CRS.
(2034). It is therefore n o t necessary to identify these B. Specific optical rotation (see Tests).
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): A , B, TESTS
C, D, E, F, G. Specific optical rotation (2.2.7)
+ 53 to + 57 (dried substance).
Dissolve 0.250 g in anhydrous ethanol R and dilute to
T yr-P he— X— Y— C ys-Pro— Z—Gly-NH2 25.0 m L with the same solvent.
4 i 8
Related substances
Liquid chromatography (2.2.29).
A. X = Gin, Y = Asp, Z = D-Arg: [5-L-aspartic
a d d ] desmopressin, Test solution Dissolve 20.0 mg o f the substance to be
examined in 25 m L of acetomtrUe R1 an d dilute to 50.0 m L
B. X = Glu, Y = A sn, Z = D-Arg: [4-L-glutamic
with water R.
a d d ] desm opressin,
Reference solution (a) Dissolve 4 mg o f desogestrelfor system
D . X = Gin, Y = Asn, Z = L-Arg: [8-l-
suitability CRS (containing impurities A, B, C and D) in
arginine] desmopressin,
5 m L o f acetonitrüe R1 and dilute to 10.0 m L w ith water R.
1-702 Desogestrel 2016
Reference solution (b) D ilute 1.0 m l. o f the test solution to Calculate the percentage content o f C 22H 30O from the areas
100.0 m L with a mixture o f equal volumes of acetonitrile R1 of the peaks and the declared content o f desogestrel CRS.
and water R.
IM P U R IT IE S
Reference solution (c) Dilute 1.0 m L of reference solution (b) Specified impurities A, B, C, D
to 10.0 m L with a mixture of equal volumes of acetonitrik R l
Other detectable impurities (the following substances would, if
and water R.
present at a sufficient level, be detected by one or other of
Reference solution (d) Dissolve 20.0 mg of desogestrel CRS in the tests in the monograph. T hey are limited by the general
25 m L of acetonitrik R l and dilute to 50.0 m L with water R. acceptance criterion for other/unspecified impurities and/or
Column: by the general monograph Substances for pharm aceutical use
— size. I = 0.25 m , 0 = 4.6 mm, (2034). It is therefore not necessary to identify these
— stationary phase: sterically protected octadecylsUyl silica gel impurities for demonstration of compliance. See also 5.10.
for chromatography R (5 fim), Control of impurities in substances for pharmaceutical use): E.
— temperature: 50 °C.
Mobile phase water R, acetonitrik R l (27:73 ViV). CH
Reference solution (b) Dissolve 10 mg of cortisone acetate R in Retention time Betamethasone 17-valerate = about 7.5 min;
reference solution (a) and dilute to 10 m L with reference desoxycortone acetate = about 9.5 min.
solution (a). System suitability: reference solution (a):
— resolution: minimum 4.5 between the peaks due to
Plate TLC silica gel F254 plate R.
betam ethasone 17-valerate and desoxycortone acetate;
Mobile phase Add a mixture of 1.2 volumes o f water R and if necessary, adjust the concentration o f acetonitrile in the
8 volumes o f methanol R to a mixture of 15 volumes of mobile phase.
ether R and 77 volumes o f methylene chloride R.
Limits:
Application 5 |iL. — unspecified impurities: for each impurity, not more than
Development Over 2/3 o f the plate. 0.2 times the area of the principal peak in the
Drying In air. chromatogram obtained with reference solution (b)
Detection A Examine in ultraviolet light at 254 nm. ( 0.10 p er cent);
— total: n o t more than the area of the principal peak in the
Results A T he principal spot in the chromatogram obtained
chromatogram obtained with reference solution (b)
w ith the test solution is similar in position and size to the
(0.5 p er cent);
principal spot in the chromatogram obtained with reference
— disregard limit: 0.1 times the area o f the principal peak in
solution (a).
the chromatogram obtained with reference solution (b)
(0.05 p er cent).
1-704 Dexamethasone 2016
F. 9-fluoro-l lß ,21-dihydroxy-16a-methylpregna-l,4-diene-
3,20-dione,
A c tio n a n d u se
Glucocorticoid.
G. 9-fluoro-l 1ß, 17-dihydroxy-16a-methyl-3,20-dioxopregna-
l,4-dien-21-yl acetate (dexamethasone acetate), PhEur__________________________________________________________
D E F IN IT IO N
9-Fluoro-l 1P, 17-dihydroxy-16a-methyl-3,20-dioxopregna-
l,4-dien-21-yl acetate.
C o n te n t
97.0 per cent to 103.0 per cent (dried substance).
CHARACTERS
A p p e a ra n c e
W hite or alm ost white, crystalline powder.
H. 17-hydroxy-16a-m ethyl-3,20-dioxopregna-l,4,9(l l)-trien-
21-yl acetate, S o lu b ility
Practically insoluble in water, freely soluble in ethanol
(96 per cent), slightly soluble in methylene chloride.
It shows polymorphism (5.9).
ID E N T IF IC A T IO N
First identification B, C.
Second identification A , C, D, E, F.
A. Dissolve 10.0 m g in anhydrous ethanol R and dilute to
100.0 m L with the same solvent. Place 2.0 m L of this
I. 9a,lla-epoxy-17,21-dihydroxy-16a-m ethylpregna-l,4- solution in a ground-glass-stoppered tube, add 10.0 m L o f
diene-3,20-dione, phenylhydrazine-sidfitric acid solution R, mix and heat in a
water-bath at 60 °C for 20 m in. Cool immediately.
T he absorbance (2.2.25) m easured at the absorption
m aximum at 419 nm is no t less than 0.35.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison dexamethasone acetate CRS.
If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and the reference
substance separately in methylene chloride R, evaporate to
J. 17,21-dihydroxy-16a-m ethylpregna-l,4-diene-3,l 1, 20 -
dryness and record new spectra using the residues.
trione,
C. Thin-layer chromatography (2.2.27).
Solvent mixture methanol R, methylene chloride R (1:9 V/V).
Test solution Dissolve 10 m g o f the substance to be examined
in the solvent mixture and dilute to 10 m l. with the solvent
mixture.
2016 Dexamethasone Acetate 1-707
Reference solution (a) Dissolve 20 mg of dexamethasone Reference solution (b) Dilute 1.0 m L o f the test solution to
acetate CRS in the solvent mixture and dilute to 20 m l. with 100.0 m L with the mobile phase. Dilute 1.0 m L of this
the solvent mixture. solution to 10.0 m L with the mobile phase.
Reference solution (b) Dissolve 10 mg o f cortisone acetate R in Reference solution (c) Dissolve the contents of a vial of
reference solution (a) and dilute to 10 m L with reference dexamethasone acetate impurity E CRS in 1.0 m L o f the
solution (a). mobile phase.
Plate TLC silica gel F 254 plate R. Column:
Mobile phase Add a mixture of 1.2 volumes of water R and — size. I = 0.25 m , 0 = 4.6 mm;
8 volumes of methanol R to a mixture o f 15 volumes of — stationary phase: octadecylsilyl silica gel for chromatography R
ether R and 77 volumes of methylene chloride R. (5 |im).
Application 5 |iL. Mobile phase Mix 380 m L of acetonitrile R with 550 m L of
water R and allow to equilibrate; dilute to 1000.0 m L with
Development Over 3/4 of the plate.
water R and mix again.
Drying In air.
Flow rate 1 mL/min.
Detection A Examine in ultraviolet light at 254 nm.
Detection Spectrophotometer at 254 nm.
Results A T he principal spot in the chromatogram obtained
Injection 20 |iL.
with the test solution is similar in position and size to the
principal spot in the chromatogram obtained with reference Run time 2.5 times the retention time of dexamethasone
solution (a). acetate.
Detection B Spray with alcoholic solution of sulfuric add R, heat Identification of impurities Use the chromatogram obtained
at 120 °C for 10 min or until the spots appear, and allow to with reference solution (a) to identify the peaks due to
cool; examine in daylight and in ultraviolet light at 365 nm . impurities A and D; use the chromatogram obtained with
reference solution (c) to identify the peak due to impurity E.
Residts B T he principal spot in the chromatogram obtained
with the test solution is similar in position, colour in daylight, Relative retention W ith reference to dexamethasone ^cetate
fluorescence in ultraviolet light at 365 nm and size to the (retention time = about 22 min): impurity A = about 0.4;
principal spot in the chromatogram obtained with reference impurity D = about 0.9; impurity E = about 1.2.
solution (a). System suitability, reference solution (a):
System suitability, reference solution (b): — resolution: minimum 3.3 between die peaks due to
— the chromatogram shows 2 clearly separated spots. impurity D and dexamethasone acetate.
D . Add about 2 mg to 2 m L of sulfuric add R and shake to Limits:
dissolve. W ithin 5 min, a faint reddish-brown colour — impurity D: n o t more than 3 times the area o f the
develops. Add this solution to 10 m L o f water R and mix. principal peak in the chromatogram obtained with
T he colour is discharged and a clear solution remains. reference solution (b) (0.3 per cent);
— impurities A , E: for each impurity, not more than twice
E. Mix about 5 mg with 45 mg of heavy magnesium oxide R
the area of the principal peak in the chromatogram
and ignite in a crucible until an almost white residue is
obtained with reference solution (b) (0.2 per cent);
obtained (usually less than 5 min). Allow to cool, add 1 m L
— unspecified impurities: for each impurity, n o t more than the
of water R, 0.05 m L of phenolphthalein solution R1 and about
area of the principal peak in the chromatogram obtained
1 m L of dilute hydrochloric add R to render the solution
with reference solution (b) (0.10 per cent);
colourless. Filter. T o a freshly prepared mixture of 0.1 m L of
— total: not m ore than 5 times the area of the principal peak
alizarin S solution R and 0.1 m L of zirconyl nitrate solution R,
in the chromatogram obtained with reference solution (b)
add 1.0 m L o f the filtrate. Mix, allow to stand for 5 min and
(0.5 per cent);
compare the colour o f the solution with that of a blank
— disregard limit. 0.5 times the area of the principal peak in
prepared in the same manner. The test solution is yellow and
the chromatogram obtained with reference solution (b)
the blank is red.
(0.05 per cent).
F. About 10 mg gives the reaction o f acetyl (2.3.1).
Loss o n d ry in g (2.2.32)
TESTS M aximum 0.5 per cent, determined on 0.500 g by drying
S pecific o p tical ro ta tio n (2.2.7) in vacuo in an oven at 105 °C.
+ 94 to + 99 (dried substance).
ASSAY
Dissolve 0.250 g in anhydrous ethanol R and dilute to Dissolve 0.100 g in ethanol (96 per cent) R and dilute to
25.0 m L with the same solvent. 100.0 m L with the same solvent. Dilute 2.0 m L o f this
R e la te d su b sta n c e s solution to 100.0 m L with ethanol (96 per cent) R. Measure
Liquid chromatography (2.2.29). Carry out the test protected the absorbance (2.2.25) at the absorption maximum at
from light. 238.5 nm.
Test solution Dissolve 25 mg of the substance to be examined Calculate the content o f C 24H 31F 0 6 taking the specific
in about 4 m L of acetonitrile R and dilute to 10.0 m L with absorbance to be 357.
water R. STORAGE
Reference solution (a) Dissolve 2 mg o f dexamethasone CRS Protected from light.
(impurity A) and 2 m g of betamethasone acetate CRS
(impurity D) in 100.0 mT. o f the mobile phase and sonicate IMPURITIES
for about 10 m in (solution A). Mix 6.0 m L o f the test Specified impurities A, D , E
solution and 1.0 mT. o f solution A and dilute to 10.0 m L Other detectable impurities (the following substances would, if
with the mobile phase. present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
1-708 Dexamethasone Isonicotinate 2016
G. 9-fluoro-l ip-hydroxy-16a-methyl-3j20-dioxopregna-lj4-
dien- 21-yl acetate.
o o
W t-o h ch3
★* ★
★ ★
B. 14-fluoro-l ip,17-<iihydroxy-16a-methyl-3j20-dioxopregna- Dexamethasone Isonicotinate ★ ★
l }4-dien-21-yl acetate, *****
(Ph. Eur. monograph 2237)
\ 0
H CH3)
■OH CHj
PhEur_____________
D E F IN IT IO N
9-Fluoro-l 1P, 17-dihydroxy-l 6a-methyl-3,20-dioxopregna-
l,4-dien-21-yl pyridine-4-carboxylate.
D . 9-fluoro-llp,17-dihydroxy-16p-m ethyl-3,20-dioxopregna- Content
l,4-dien-21-yl acetate (betamethasone acetate), 99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
Appearance
H CH \ v --(
-o h 0 ^ W hite or alm ost white crystalline powder.
CHj
Solubility
Practically insoluble in water, slighdy soluble in anhydrous
ethanol and in acetone.
ID E N T IF IC A T IO N
E. 9-fluoro- 11 P, 17-dihydroxy-16a-methyl-3,20-dioxopregn-4- Infrared absorption spectrophotometry (2.2.24).
en- 21 -yl acetate. Comparison dexamethasone isonicotinate CRS.
TESTS
Specific optical rotation (2.2.7)
+ 142 to + 146 (dried substance).
Suspend 0.200 g in 4.0 m l. o f ethyl acetate R and dilute to
20.0 m L w ith ethanol (96 per cent) R. T reat in an ultrasonic
bath until a clear solution is obtained.
Related substances
F. 17-hydroxy-16a-methyl-3,20-dioxo-9P,l ip-epoxypregna- Liquid chrom atography (2.2.29). Prepare solutions immediately
l,4-dien-21-yl acetate, before use.
2016 Dexamethasone Sodium Phosphate 1-709
25-35 68 32
B. 9-fluoro-l 1ß, 17-dihydroxy-l6a-methyl-3,20-dioxopregna-
l,4-dien-21-yl acetate (dexamethasone acetate),
Flow rate 1.2 mlVmin.
Detection Spectrophotom eter at 240 nm .
Injection 10 jjJL.
Identification of impurities Use the chromatogram supplied
with dexamethasone isomcotinate for impurity C
identification CRS and the chromatogram obtained with
reference solution (c) to identify the peak due to impurity C.
Relative retention W ith reference to dexamethasone C. 9-fluoro-llß , 17-dihydroxy-16a-methylpregna-l,4-diene-
isonicotinate (retention time = about 12 min):
3,20-dione (21-deoxydexamethasone).
impurity A = about 0.4; impurity C = about 0.6;
impurity B = about 0.8. _________________________________________________________ PhEur
Reference solution (c) Dilute 1.0 mL of the test solution to Dissolve 50 mg in water R and dilute to 100 m L with the
100.0 m L with mobile phase A. D ilute 1.0 m L of this same solvent. T o 10 m L of this solution add 5 m L of
solution to 10.0 m L with mobile phase A. molybdovanadic reagent R, mix and allow to stand for 5 min.
Column: Any yellow colour in the solution is not m ore intense than
— sizer. I = 0.125 m, 0 = 4.6 mm; that in a standard prepared at the same time in the same
— stationary phase, end-capped, octylsifyl silica gel for m anner using 10 m L o f phosphate standard solution (5 ppm
chromatography R (5 pm); POJ R.
— temperature: 30 °C. E th a n o l (2.4.24, System A)
Mobile phase. M aximum 1.5 per cent.
— mobile phase A: mix 300 mT. of solution A and 350 mT. o f W a te r (2.5.12)
water R, adjust to p H 3.8 with acetic acid R, then add M aximum 10.0 per cent, determined on 0.200 g.
350 m L of methanol R;
— mobile phase B: adjust 300 m L of solution A to p H 4.0 A SSAY
with acetic acid R, then add 700 m L of methanol R; Liquid chromatography (2.2.29).
Test solution Dissolve 30.0 mg of the substance to be
examined in the mobile phase and dilute to 50.0 m L with
Time Mobile phase A Mobile phase B
(min)
the mobile phase. Dilute 5.0 mL o f the solution to 50.0 m L
(per cent V/V) (per cent V/V)
0 -3 .5
with the mobile phase.
90 10
Reference solution (a) Dissolve 2 m g of dexamethasone CRS
3.5-23.5 90 -»60 10 -» 40
(impurity A) and 2 mg of dexamethasone sodium
23.5 - 34.5 60 -»5 40 -» 95 phosphate CRS in 2 m L o f tetrahydrofuran R, then dilute to
34.5 - 50 5 95 100.0 m L with the mobile phase. Dilute 5.0 m L of this
solution to 50.0 m L with the mobile phase.
Reference solution (b) Dissolve 30.0 mg of dexamethasone
Flow rate 1.0 mlVmin. sodium phosphate CRS in the mobile phase and dilute to
Detection Spectrophotometer at 254 nm . 50.0 m L with the mobile phase. Dilute 5.0 m L o f the
Injection 20 |±L. solution to 50.0 m L with the mobile phase.
Identification of impurities Use the chromatogram supplied Column:
with dexamethasone sodium phosphate for peak — size. I = 0.15 m , 0 = 4.6 mm;
identification CRS and the chromatogram obtained with — stationary phase: end-capped octadecylsUyl silica gel for
reference solution (b) to identify the peaks due to impurities chromatography R (7 Jim).
A, C, D , E , F and G; use the chromatogram obtained with Mobile phase Mix 520 m L of water R with 2 m L of phosphoric
reference solution (a) to identify the peak due to impurity B. add R. Adjust the temperature to 20 °C, then adjust to
Relative retention W ith reference to dexamethasone sodium p H 2.6 with sodium hydroxide R. M ix this solution with
phosphate (retention time = about 22 min): 36 m L o f tetrahydrofuran R and 364 mL of methanol R.
im purity C = about 0.5; impurity D = about 0 . 6; Flow rate 1.5 mlVmin.
impurity E = about 0.8; impurity F = about 0.92; Detection Spectrophotometer at 254 nm.
im purity B = about 0.95; impurity A = about 1.37; Injection 20 |±L.
impurity G = about 1.41.
Run time 3 times the retention tim e of dexamethasone
System suitability,: reference solution (a): sodium phosphate.
— resolution: minimum 2.0 between the peaks due to
Identification of impurities Use the chromatogram obtained
impurity B and dexamethasone sodium phosphate.
with reference solution (a) to identify the peak due to
Limits'. impurity A.
— correction factor, for the calculation of content, multiply the
peak area o f impurity A by 0.75;
Relative retention W ith reference to dexamethasone sodium
— impurity A: not more than 5 times the area o f the phosphate (retention time = about 8 min):
impurity A = about 2.0.
principal peak in the chromatogram obtained with
reference solution (c) (0.5 per cent); System suitability: reference solution (a):
— impurity G: n o t more than 3 times the area o f the — resolution: minimum 6.0 between the peaks due to
principal peak in the chromatogram obtained with dexamethasone sodium phosphate and im purity A.
reference solution (c) (0.3 per cent); Calculate the percentage content o f C22H28FNa20 gP using
— impurities B, C, D, E, F: for each impurity, no t m ore than the chromatogram obtained with reference solution (b) and
twice the area of the principal peak in the chromatogram taking into account the assigned content o f dexamethasone
obtained with reference solution (c) (0.2 per cent); sodium phosphate CRS.
— unspecified impurities: for each impurity, n o t more than the STORAGE
area o f the principal peak in the chromatogram obtained
In an airtight container, protected from light.
with reference solution (c) (0.10 per cent);
— total: n o t more than 10 times the area o f the principal IM P U R IT IE S
peak in the chromatogram obtained with reference Spedfied impurities A, B, C, D, E, F , G
solution (c) ( 1.0 per cent); Other detectable impurities (the following substances would, if
— disregard limit: 0.5 times the area o f the principal peak in present at a sufficient level, be detected by one or other of
the chromatogram obtained with reference solution (c) the tests in the monograph. They are limited by the general
(0.05 per cent). acceptance criterion for other/unspecified impurities and/or
In o rg a n ic p h o sp h a te s by the general monograph Substances for pharmaceutical use
M axim um 1 per cent. (2034). It is therefore n o t necessary to identify these
1-712 Dexamfetamine Sulfate 2016
C n a r^ O H
.HjSCU
A ’CHs tt Me
)
X (C 9H 13N ) 2,H 2S 0 4 368.5 51-63-8
A. 9 -flu o ro -lip , 17,21 -trihydroxy-16a-methylpregna-1,4-
A c tio n a n d u se
diene-3,20-dione (dexamethasone),
Amfetamine.
P r e p a r a tio n
D e x a m f e t a m in e Tablets
D E F IN IT IO N
Dexamfetamine Sulfate is (5)-a-m ethylphenethylam ine
sulfate. It contains not less th an 9 9 .0 % and not m ore than
1 0 0 .5 % o f (C 9H 13N ) 2JH 2S 04 , calculated with reference to
the dried substance.
B. 9-fluoro-l ip,17-dihydroxy-16p-methyl-3,20-dioxopregna-
l,4-dien-21-yl dihydrogen phosphate (betamethasone C H A R A C T E R IS T IC S
phosphate), A white or almost white, crystalline powder.
F red y soluble in water, slightly soluble in ethanol (96%) \
practically insoluble in ether.
ID E N T IF IC A T IO N
A. Dissolve 1 g in 5 0 m L of water, add 10 m L of 5m sodium
hydroxide and 0 .5 m L of benzoyl chloride and shake. Repeat
the addition o f benzoyl chloride in 0 .5 m L quantities until no
further predpitate is produced. T h e melting point o f the
predpitate, after recrystallising twice from ethanol (50%), is
about 157°, Appendix V A.
B. Dissolve 2 mg in 4 m L o f water, add 1 m L of
1m hydrochloric add, 2 m L o f diazotised nitroanHine solution,
4 m L of 1m sodium hydroxide and 2 m L o f butan-l-ol, shake
and allow to separate. A red colour is produced in the
butanol layer (distinction from m ethylam fetamine).
C, D , E, F. for each impurity, one or more C. Yields the reactions characteristic of sulfates, Appendix VI.
diastereoisomer(s) o f (9-fluoro-11(3,17a-dihydroxy-16-methyl-
3.17 -dioxo-D -hom o-androsta-1,4-dien-17 a-yl)methyl TESTS
dihydrogen phosphate (undefined stereochemistry at C-16 A c id ity o r alk alin ity
and C-17a), o r (9-fluoro-11 p, 17-dihydroxy-16a-methyl- Dissolve 0 .5 g in 10 m L o f water and titrate with
3.17 a-dioxo-D -hom o-androsta-1,4-dien-17-yl)methyl 0 .0 1 m hydrochloric add FS o r 0 .0 1 m sodium hydroxide VS
dihydrogen phosphate (undefined stereochemistry at C -17), using methyl red solution as indicator. N o t m ore than 0.1 m L
o f 0.01m hydrochloric add VS or 0.01m sodium hydroxide VS is
required to change the colour o f the solution.
S p ecific o p tic a l ro ta tio n
In an 8 .0 % w/V solution, + 1 9 .5 to + 2 2 .0 , calculated with
reference to the dried substance, Appendix V F.
L oss o n d ry in g
W hen dried to constant w dght at 105°, loses not m ore than
G. 9-fluoro- 11P, 17-dihydroxy-16a-m ethyl-3-oxoandrosta-1,4- 1 .0 % of its weight. Use 1 g.
diene-17P-carboxylic ad d , S u lfa te d a s h
N o t more than 0.1% , Appendix IX A.
A SSA Y
Dissolve 0 .4 g in 1 2 0 m L o f water, add 2 m L of 5m sodium
hydroxide and distil into 5 0 rnT. of 0 .1 m hydrochloric add KS,
continuing the distillation until only 5 m L o f liquid is left in
the distillation flask. T itrate the excess o f a d d with
0.1m sodium hydroxide KS using methyl red solution as
indicator. Each m l. o f 0.1m hydrochloric add KS is equivalent
H . 9-fluoro-l ip , 17-dihydroxy-16a-methyl-3,20-dioxopregn- to 1 8 .4 2 m g of (C 9H 13N ) 2,H 2S 0 4 .
4-en-21-yI dihydrogen phosphate.
PhEur
2016 Dexchlorphenirainine Maleate 1-713
★ ★ A p p e a ra n c e o f so lu tio n
Dexchlorpheniramine Maleate ★ ★ Solution S is clear (2.2.1)- and not more intensely coloured
★ . ★
(Ph. Eur. monograph 1196) *★* than reference solution BY 6 (2.2.2, Method II).
p H (2.2.3)
4.5 to 5.5.
c o 2h Dissolve 0.20 g in 20 m L of water R.
( c o 2h
S pecific o p tic a l ro ta tio n (2.2.7)
+ 22 to + 23 (dried substance), determined on solution S.
R e la te d su b sta n c e s
Gas chromatography (2.2.28).
2438-32-6 Test solution Dissolve 10.0 mg of the substance to be
examined in 1.0 m L of methylene chloride R.
A c tio n a n d u se Reference solution Dissolve 5.0 mg o f brompheniramine
Histam ine H I receptor antagonist; antihistamine. maleate CRS in 0.5 m L of methylene chloride R and add
0.5 m L of the test solution. Dilute 0.5 m L o f this solution to
PhEur.
50.0 m L with methylene chloride R.
D E F IN IT IO N Column:
(3S)-3-(4-Chlorophenyl)-N,N-dimethyl-3-(pyridin-2- — material: glass;
yl) propan- 1-amine (Z)-butenedioate. — sizer. I = 2.3 m , 0 = 2 mm;
C o n te n t — stationary phase, ad d - and base-washed silanised
98.0 per cent to 100.5 per cent (dried substance). diatomaceous earth for gas chromatography R (135-175 |im)
impregnated with 3 per cent mlm of a mixture of
CHARACTERS
50 per cent of poly(dimethyl)siloxane and 50 per cent of
A p p e a ra n c e
poly(diphenyl)siloxane.
W hite or almost white, crystalline powder.
Carrier gas nitrogen for chromatography R.
S o lu b ility
Flow rate 20 mlVmin.
Very soluble in water, freely soluble in ethanol (96 per cent),
in m ethanol and in methylene chloride. Temperature:
— column: 205 °C;
ID E N T IF IC A T IO N — infection port and detector. 250 °C.
First identification A , C, E Detection Flame ionisation.
Second identification A , B, D, E Iiyection 1 jiL.
A. Specific optical rotation (see Tests). Run time 2.5 times the retention time of
B. Melting point (2.2.14): 110 °C to 115 °C. dexchlorpheniramine.
C . Infrared absorption spectrophotometry (2.2.24). System suitability: reference solution:
Preparation Discs of potassium bromide R. — resolution: minimum 1.5 between the peaks due to
Comparison dexchlorpheniramine maleate CRS. dexchlorpheniramine and brompheniramine.
D . Thin-layer chromatography (2.2.27). Limits:
— impurity A: n o t more than 0.8 times the area of the peak
Test solution Dissolve 0.10 g of the substance to be examined
due to dexchlorpheniramine in the chromatogram
in methanol R and dilute to 5.0 m L with the same solvent.
obtained with the reference solution (0.4 per cent);
Reference solution Dissolve 56 mg o f maleic acid R in — total: not m ore than twice the area of the peak due to
methanol R and dilute to 10 m L with the same solvent. dexchlorpheniramine in the chromatogram obtained with
Plate TLC silica gel F2 5 4 plate R. the reference solution (1 per cent).
Mobile phase water R, anhydrous formic acid R, methanol R, di E n a n tio m e ric p u rity
isopropyl ether R (3:7:20:70 V/V/V/V). Liquid chromatography (2.2.29).
Application 5 jiL. Test solution Dissolve 10.0 mg of the substance to be
Development Over a path o f 12 cm. examined in 3 m L of water R. Add a few drops of
Drying In a current o f air for a few minutes. concentrated ammonia R until an alkaline reaction is produced.
Shake with 5 m L of methylene chloride R. Separate the layers.
Detection Examine in ultraviolet light at 254 nm .
Evaporate the lower, methylene chloride layer to an oily
Results T he chromatogram obtained with the test solution residue on a water-bath. Dissolve the oily residue in
shows 2 clearly separated spots. The upper spot is similar in 2-propanol R and dilute to 10.0 m L with the same solvent.
position and size to the spot in the chromatogram obtained
Reference solution (a) Dissolve 10.0 mg o f dexchlorpheniramine
w ith the reference solution.
maleate CRS in 3 m L o f water R. Add a few drops of
E. T o 0.15 g in a porcelain crucible add 0.5 g of anhydrous concentrated ammonia R until an alkaline reaction is produced.
sodium carbonate R. H eat over an open flame for 10 min. Shake with 5 m L of methylene chloride R. Separate the layers.
Allow to cool. Take up the residue with 10 m L of dilute nitric Evaporate the lower, methylene chloride layer to an oily
acid R and filter. T o 1 m L o f the filtrate add 1 m L of residue on a water-bath. Dissolve the oily residue in
water R. T he solution gives reaction (a) of chlorides (2.3.1). 2-propanol R and dilute to 10.0 m L with the same solvent.
TESTS Reference solution (b) Dissolve 10.0 mg o f chlorphenamine
S o lu tio n S maleate CRS in 3 m L of water R. Add a few drops of
Dissolve 2.0 g in water R and dilute to 20.0 m L with the concentrated ammonia R until an alkaline reaction is produced.
sam e solvent.
1-714 Dexpanthenol 2016
p H (2.2.3)
T he p H of solution S is not greater than 10.5.
Dextran 1 for Injection *****
+*
S p ecific o p tic a l ro ta tio n (2.2.7) (Ph. Eur. monograph 1506) *
T he specific optical rotation is + 29.0 to + 32.0, determined
on solution S and calculated with reference to the anhydrous A ctio n a n d u se
substance. Plasma substitute.
3 -A m in o p ro p an o l PhEur_________________________________________________________
Examine by thin-layer chromatography (2.2.27), using silica
D E F IN IT IO N
gel G R as the coating substance.
Low-molecular-weight fraction o f dextran, consisting of a
Test solution (a) Dissolve 0.25 g of the substance to be mixture o f isomaltooligosaccharides.
examined in anhydrous ethanol R and dilute to 5 m L with the
Average relative molecular mass About 1000.
same solvent.
Test solution (b) Dilute 1 m L of test solution (a) to 10 m L P R O D U C T IO N
with anhydrous ethanol R. It is obtained by hydrolysis and fractionation of dextrans
Reference solution (a) Dissolve the contents of a vial of produced by fermentation of sucrose using Leuconostoc
dexpanthenol CRS in 1.0 m L of anhydrous ethanol R to obtain mesenteroides strain N R R L B-512 = C IP 78.59 or substrains
a concentration of 5 mgfrnL. thereof (for example L. mesenteroides B-512 F = N C T C
10817).
Reference solution (b) Dissolve 25 m g o f 3-aminopropanol R in
anhydrous ethanol R and dilute to 100 m L with the same It is prepared in conditions designed to minimise the risk of
microbial contamination.
solvent.
Apply separately to the plate 10 pL of each solution. Develop CHARACTERS
over a path of 15 cm using a mixture of 20 volumes of A p p e a ra n ce
concentrated ammonia R, 25 volumes of methanol R and White or almost white hygroscopic powder.
55 volumes of butanol R. Allow the plate to dry in air, spray S olub ility
with a 100 g/L solution o f trichloroacetic acid R in methanol R Very soluble in water, very slightly soluble in ethanol
and heat at 150 °C for 10 min. Spray with a 1 g/L solution (96 per cent).
of ninkydrin R in methanol R and heat at 120 °C until a
ID E N T IF IC A T IO N
colour appears. Any spot due to 3-am inopropanol in the
chromatogram obtained with test solution (a) is n o t more A. Dissolve 3.000 g in water R, heat on a water-bath and
intense than the spot in the chromatogram obtained with dilute to 100.0 m L with the same solvent. The specific
reference solution (b) (0.5 per cent). optical rotation (2.2.7) is + 148 to + 164, calculated with
reference to the dried substance. Dry an aliquot of the
H eav y m e ta ls (2.4.8) solution first on a water-bath and then to constant weight
12 m L o f solution S complies with limit test A for heavy in vacuo at 70 °C. Calculate the dextran content after
metals (20 ppm). Prepare the reference solution using lead correction for the content of sodium chloride.
standard solution (1 ppm Pb) R.
B. Infrared absorption spectrophotometry (2.2.24).
W a te r (2.5.12)
Preparation T o 1-2 m g add 1 or a few drops of water R.
N ot more than 1.0 per cent, determined on 1.000 g.
Grind in an agate m ortar for 1-2 min. Add about 300 m g of
S u lfa te d a sh (2.4.14) potassium bromide R and mix to a slurry b u t do not grind.
N ot m ore than 0.1 per cent, determined on 1.0 g. Dry in vacuo at 40 °C for 15 min. Crush the residue. If it is
A SSA Y not dry, dry for another 15 min. Prepare a disc using
T o 0.400 g add 50.0 m L of 0.1 Mperchloric acid. Boil under potassium bromide R.
a reflux condenser for 5 h protected from humidity. Allow to Comparison Repeat the operations using dextran 1 CRS.
cool. Add 50 m L of dioxan R by rinsing the condenser, Blank Run the infrared spectrum with a blank disc using
protected from hum idity. Add 0.2 m L o f naphtholbenzein potassium bromide R in the reference beam.
solution R and titrate with 0.1 M potassium hydrogen phthalate C. Molecular-mass distribution (see Tests).
until the colour changes from green to yellow. Carry out a
blank titration. TESTS
S o lu tio n S
1 m L of 0.1 M perchloric acid is equivalent to 20.53 mg
Dissolve 7.5 g in carbon dioxide-free water R, heat on a water-
of C 9 H 19 NO 4 .
bath and dilute to 50 m L with the same solvent
STO RA G E
A b so rb an ce (2.2.25)
In an airtight container. M aximum 0.12, determined at 375 a m on solution S.
PhEur A cidity o r alk alin ity
T o 10 m L of solution S add 0.1 m L o f phenolphthalein
solution R. The solution is colourless. Add 0.2 m L of 0.01 M
sodium hydroxide. The solution is pink. Add 0.4 m L of
0.01 M hydrochloric acid. The solution is colourless.
Add 0.1 m L o f methyl red solution R. T h e solution is red or
orange.
N itro g e n -c o n ta in in g su b sta n c e s
M aximum 110 ppm o f N .
Carry out the determination o f nitrogen by sulfuric acid
digestion (2.5.9), using 0.200 g and heating for 2 h. Collect
1-716 Dextran 40 2016
PhEur_________________________________________________________ _
D E F IN IT IO N
M ixture of polysaccharides, principally of the a - l , 6-glucan
type.
Average relative molecular mass A bout 40 000.
2016 Dextran 60 1-717
Comparison dextran CRS. is not greater than 185 000. The average molecular mass of
C. Molecular-mass distribution (see Tests). the 10 per cent low fraction is not less than 15 000.
Introduce 0.50 g to 30 m L o f a freshly prepared 74.6 g/L hydroxide solution R and, dropwise with shaking, 0.5 m L o f
solution of potassium chloride R. Allow to stand for 2 min. copper sulfate solution R and boil. A red p redpitate is
Determ ine the p H on the mucilage obtained. produced.
B o ro n c . It is very soluble in boiling water R, forming a
M aximum 30 ppm . mucilaginous solution.
Inductively coupled plasma-atomic emission spectrometry TESTS
(ICP-AES) (2.2.57). p H (2.2.3)
Test solution Introduce 3.0 g into a platinum dish and 2.0 to 8 .0 .
m oisten with 5 m L o f a 32.1 g/L solution of magnesium Disperse 5.0 g in 100 m L o f carbon dioxide-free water R.
nitrate R ina, mixture o f equal volumes o f ethanol
C h lo rid e s
(96 per cent) R and distilled water R. Evaporate to dryness on
M axim um 0.2 per c e n t
a water-bath. Ignite a t 550 °C for 5 h. Take up the residue
with 5 m L of 6 M hydrochloric acid R and transfer to a 50 m L Dissolve 2.5 g in 50 m L o f boiling water R, dilute to 100 m L
volumetric flask. A dd about 20 m L o f distilled water R and with water R and filter. Dilute 1 m L o f the filtrate to 15 mL,
allow to digest for 1 h on a w ater-bath. Allow to cool and add 1 m L o f dilute nitric add R, p our the mixture as a single
dilute to 50.0 m L with distilled water R. addition into 1 m L o f silver nitrate solution R2 and allow to
stand for 5 m in protected from light. W hen viewed
Reference solutions Prepare the reference solutions using a transversely against a black background any opalescence
solution o f boric acid R containing 10 ppm of boron. Proceed
produced is n o t more intense than th at obtained by treating a
as described for the test solution.
mixture o f 10 m L o f chloride standard solution (5 ppm Cl) R
Wavelength 249.773 nm. and 5 m L o f water R, prepared in the same m anner.
H eav y m e ta ls (2.4.8) R e d u c in g su g a rs
M axim um 30 ppm . Maximum 10 per cent, calculated as glucose C ôH^O tì.
1.0 g complies with test F. Prepare the reference solution T o a quantity of dextrin equivalent to 2.0 g (dried substance)
using 3 m L o f lead standard solution (10 ppm Pb) R. add 100 m L o f water R, shake for 30 min, dilute to
L oss o n d ry in g C2.2.32) 200.0 m L with water R and filter. T o 10.0 m L o f alkaline
M axim um 10.0 per cent, determ ined on 1.000 g by drying in cupri-tartaric solution R add 20.0 m L o f the filtrate, mix, and
an oven at 105 °C for 15 h. heat on a h o t plate adjusted to bring the solution to boil
S u lfa te d a sh (2.4.14) within 3 m in. Boil for 2 min, and cool immediately.
M aximum 0.4 p er cent, determined on 1.0 g. Add 5 m L o f a 300 g/L solution o f potassium iodide R and
10 m L o f 1 M sulfuric add, mix, and titrate immediately with
M ic ro b ia l c o n ta m in a tio n 0.1 M sodium thiosulfate, using starch solution R, added
TA M C : acceptance criterion 10 2 C FU /g (2.6.12), towards the end of the titration, as indicator. R epeat the
determ ined using the pour-plate method. procedure beginning with “T o 10.0 m L o£..”, using, in place
___________________________________________________________ PhEur of the filtrate, 20.0 m L o f a 1 g/L solution of glucose R,
accurately prepared. Perform a blank titration. (Vb — VÙ) is
not greater than (VB — v$)i in which Vjs, Vu and Vs are the
num ber o f millilitres of 0.1 M sodium thiosulfate consum ed in
the titrations o f the blank, the dextrin and the glucose,
Dextrin ***** respectively.
(Ph. Eur. monograph 1507) * H eav y m e ta ls (2.4.8)
M axim um 20 ppm .
A ctio n a n d u se
1.0 g complies with test c. Prepare the reference solution
Excipient.
using 2 m L o f lead standard solution (10 ppm Pb) R.
PhEur___________________________________________________________ L oss o n d ry in g (2.2.32)
M aximum 13.0 per cent, determined on 1.000 g by drying at
D E F IN IT IO N
130-135 ° c for 90 min.
M aize, potato or cassava starch partly hydrolysed and
modified by heating with or w ithout the presence o f adds, S u lfa te d a s h (2.4.14)
alkalis or pH -control agents. M axim um 0.5 per cent, determined on 1.0 g.
CHARACTERS F U N C T IO N A L IT Y -R E L A T E D C H A R A C T E R IS T IC S
A p p e a ra n c e This section provides information on characteristics that are
W hite or alm ost white, free-flowing powder. recognised as bâng relevant control parameters for one or more
functions of the substance when used as an excipient (see chapter
S o lu b ility
5.15). This section ừ a non-mandatory part of the monograph
Very soluble in boiling water forming a mucilaginous
and ừ is not necessary to verify the characteristics to demonstrate
solution, slowly soluble in cold water, practically insoluble in
compliance. Control of these characteristics can however contribute
ethanol (96 per cent).
to the quality of a medicinal product by improving the consistency
ID E N T IF IC A T IO N of the manufacturing process and the performance of the medicinal
A. Suspend 1 g in 50 m L o f water R, boil for 1 min and product during use. Where control methods are cited, they are
cool. T o 1 m L o f the solution add 0.05 m L o f iodine recognised as being suitable for the purpose, but other methods can
solution R l. A dark blue or reddish-brown colour is also be used Wherever results for a particular characteristic are
produced, which disappears o n heating. reported, the control method must be indicated
B. Centrifuge 5 m L o f the mucilage obtained in identification The following characteristics may be relevant for dextrm used as
test A. T o the upper layer add 2 m L o f dilute sodium filler and binder, in tablets and capsules.
2016 Dextromethorphan Hydrobromide 1-721
P a rtic le -s iz e d istrib u tio n (2.9.31) Test solution Dissolve 25 mg of the substance to be examined
or 2.9.38). in methanol R and dilute to 10 m L with the same solvent.
P o w d e r flow (2.9.36) Reference solution Dissolve 25 mg o f dextromethorphan
The following characteristic may be relevant for dextrin used as hydrobromide CRS in methanol R and dilute to 10 m L with
viscosity-increasing agent. the same solvent.
A p p a re n t v iscosity (2.2.10) Plate TLC silica gel G plate R.
Typically 100 mPa-s to 350 mPa-s (dried substance), Mobile phase concentrated ammonia R, methylene chloride R,
depending on the grade of dextrin. methanol R, ethyl acetate R, toluene R
In a beaker, prepare a 10-50 per cent slurry so that the (2:10:13:20:55 VIVIV/VIV).
viscosity value ranges from 100 mPa-s to 350 mPa-s. Application 5 pL.
T he total mass of the sample plus water m ust be 600 g. Development Over 2/3 of the plate.
Mix with a plastic rod to obtain a homogeneous slurry. Place
Drying In air.
the beaker in a water-bath at 100 ± 1 °C. Introduce the
paddle of a stirrer into the beaker and close the beaker with a Detection Spray with potassium iodobismuthate solution R2.
lid. Start agitation at 250 r/min as rapidly as possible and Results T he principal spot in the chromatogram obtained with
carry on for exactly 30 min. Transfer the paste immediately the test solution is similar in position and size to the principal
to the beaker to be used for viscosity measurement, placed in spot in the chromatogram obtained with the reference
a water-bath at 40 ± 1 °C. Stir until the tem perature in the solution.
beaker is 40 ± 1 °C then measure the apparent viscosity D . It gives reaction (a) o f bromides (2.3.1).
using spindle no. 2 and a rotation speed of 100 r/min.
TESTS
__________________________________________________________ PhEur S o lu tio n S
Dissolve 1.0 g in ethanol (96per cent) R and dilute to 20 m l.
with the same solvent.
A p p e a ra n c e o f so lu tio n
Dextromethorphan Hydrobromide ***** Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
★ * A cid ity o r alk alin ity
(Ph. Eur. monograph 0020) *
Dissolve 0.4 g in carbon dioxide-free water R with gentle
heating, cool and dilute to 20 m L with the same solvent.
/CH,* Add 0.1 m L of methyl red solution R and 0.2 m L of 0.01 M
sodium hydroxide. T he solution is yellow. N ot more than
0.4 m L of 0.01 M hydrochloric acid is required to change the
colour of the indicator to red.
S p ecific o p tic a l ro ta tio n (2.2.7)
+ 28 to + 30 (anhydrous substance).
Dissolve 0.200 g in 0.1 M hydrochloric acid and dilute to
Ci8H 26B rN 0 ^ i20 370.3 6700-34-1 10.0 m L with the same acid.
R e la te d su b sta n c e s
A ctio n a n d u se Liquid chromatography (2.2.29).
Opioid receptor agonist; cough suppressant.
Test solution Dissolve 10.0 mg of the substance to be
PhEur__________________________________________________________ examined in the mobile phase and dilute to 10.0 m L with
the mobile phase.
D E F IN IT IO N
erzr-3-Methoxy-17 -methylinorphinan hydrobromide Reference solution (a) Dissolve 2 m g of dextromethorphan
monohydrate.
impurity A CRS in 2 m L of the test solution and dilute to
25.0 m L with the mobile phase.
C o n te n t
Reference solution (b) Dilute 1.0 m L of the test solution to
99.0 per cent to 101.0 per cent (anhydrous substance).
200.0 m L with the mobile phase.
CHARACTERS Column:
A p p e a ra n c e — size: I = 0.25 m , 0 = 4.6 mm;
A lmost white, crystalline powder. — stationary phase: octadecylsQyl silica gel for chromatography R
S o lu b ility (5 pm).
Sparingly soluble in water, freely soluble in ethanol Mobile phase Dissolve 3.11 g of docusate sodium R in a
(96 per cent). mixture of 400 m L of water R and 600 m L o f acetonitrile R,
mp add 0.56 g o f ammonium nitrate R and adjust to apparent
A bout 125 °C, with decomposition. p H 2.0 with glacial acetic acid R.
Flow rate 1.0 m lVmin.
ID E N T IF IC A T IO N
First identification A, B, D Detection Spectrophotometer at 280 nm.
Second identification A , C, D Irgection 20 pL.
A. Specific optical rotation (see Tests). Run time Twice the retention time of dextromethorphan.
B. Infrared absorption spectrophotometry (2.2.24). Relative retention W ith reference to dextromethorphan
(retention time = about 22 min): impurity B = about 0.4;
Comparison dextromethorphan hydrobromide CRS.
im purity C = about 0.8; impurity D = about 0.9;
C. Thin-layer chromatography (2.2.27). impurity A •= about 1.1.
1-722 Dextromoramide Tartrate 2016
ch3
System suitability: reference solution (a): o h n
— resolution: minimum 1.5 between the peaks due to
dextrom ethorphan and im purity A.
Limits:
— correction factor, for the calculation o f content, multiply the
peak area of im purity C by 0.2; h3c o
— impurities A , B, C, D: for each impurity, not more than
C. enr-3-methoxy-17-m ethylm orphinan-10-one,
the area o f the principal peak in the chromatogram
obtained with reference solution (b) (0.5 per cent), and ch3
(RS 092). I f the spectra are not concordant, dissolve a corresponding ratios in the chromatogram obtained with
sufficient quantity in the minimum volume of dichloromethane solution (3) (0.67% each).
IR, evaporate to dryness, dry the residue at 105° for 1 hour Sulfated ash
and prepare a new spectrum. N ot more than 0.1%, Appendix IX A.
TESTS Water
Specific optical rotation 3.0 to 5.0% w/w, Appendix IX C . Use 0.5 g.
In a 5% w/v solution in ethanol (96%), +26 to +31,
ASSAY
calculated with reference to the anhydrous substance,
T o 0.75 g add 50 m L of water, swirl to disperse, add 5 mT.
Appendix V F.
o f 5m sodium hydroxide and extract with five 25-m L quantities
Related substances o f dichloromethane, washing each extract with the same
Carry out the m ethod for gas chromatography, 20 m L o f water. Dry the combined extracts with anhydrous
Appendix IE B. Dissolve 10 mg o f triphenylamtne (internal sodium sulfate, evaporate to about 3 m L on a water bath in a
standard) in sufficient dichloromethane to produce 50 m l, current o f air and allow to evaporate to dryness at room
(solution A). tem perature. Carry out Method I for non-aqueous titration on
(1) Dissolve 0.3 g o f the substance being examined in 5 m L the residue, Appendix VUI A, using 1-naphtholbenzein solution
o f dichloromethane, add 10 m L o f water, 2 m L of as indicator. Each m l, o f 0.1m perchloric acid VS is equivalent
1.25m sodium hydroxide and 15 m L o f dichloromethane and to 54.78 mg o f C 22H 29N O 2JC j 0H 8O 3S .
shake. Extract the aqueous layer with two 20-m L quantities
o f dichloromethane. Shake the combined dichloromethane
extracts with 5 g of anhydrous sodium sulfate, filter and
evaporate to dryness at a tem perature not exceeding 40°
using a rotary evaporator. Dissolve the residue in 10 m L of Diacerein *****
★ ★
dichloromethane. * * * * *
(Ph. Eur. monograph 2409)
(2) Prepare solution (2) in the same m anner as solution ( 1)
b u t add 5 m L o f solution A to the initial solution o f the
substance being examined.
c o 2h
(3) A dd 5 m L of solution A, 10 m L o f water, 2 m L of
1.25m sodium hydroxide and 15 m L o f dichloromethane to
5 m L o f a solution in dichloromethane containing 0.022% w/v
o f (lS,2R)-l-benzyl-3-dimethylamino-2-methyl-l-phenylpropyl
acetate BPCRS and 0 .020% w/v of 4^imethylainino-3-metkyl-
l,2-diphenylbutan-2-ol hydrochloride BPCRS and shake. Extract
the aqueous layer with two 20-m L quantities of Ci9H120 8 368.3 13739-02-1
dichloromethane. Shake the combined dichloromethane
extracts with 5 g of anhydrous sodium sulfate, filter and Action and Use
evaporate to dryness at a tem perature not exceeding 40° Anti-inflammatory used in the treatm ent o f arthritis and
using a rotary evaporator. Dissolve the residue in 10 mT. of osteoarthritis.
dichloromethane.
PhEir.
CHROMATOGRAPHIC CONDITIONS
(a) Use a glass column (60 cm x 3 mm) packed with acid- DEFINITION
washed, sUanised diatomaceous support (100 to 120 mesh) 4,5-Diacetoxy-9,10-dioxo-9, 10-dihydroanthracene- 2-
coated with 3% w/w of dimethyl silicone fluid (O V-lO l is carboxylic acid.
suitable). Content
(b) Use helium as the carrier gas at 60 m L per minute. 98.0 per cent to 102.0 per cent (dried substance).
(c) Use isothermal conditions maintained at 160°. CHARACTERS
(d) Use an inlet tem perature of 150°. Appearance
(e) Use a flame ionisation detector. Yellow, crystalline powder.
Solution C M ix 25.3 volumes o f solution A and 74.6 volumes Reference solution (c) Dissolve the contents of a vial of
of solution B. If necessary, adjust to p H 9.5 using dilute diacerein impurity mixture CRS (impurities C and F) in a
sodium hydroxide solution R o r dilute sulfuric add R. mixture of 0.5 m l. of tetrahydrofuran R and 0.5 m L of the
Solution D D ilute 5 m L of dilute sulfuric add R to 500 m L solvent mixture.
with water R. Column:
Test solution Dissolve 0.100 g o f the substance to be — size: I = 0.10 m, 0 = 4.6 mm;
examined in 30 m L of solution A, mix for 10 min. — stationary phase: end-capped polar-embedded octadecylsifyl
Add 70 m L o f solution B and adjust to p H 9.5 with dilute amorphous organosUica polymer R (5 nm);
sodium hydroxide solution R or dilute sulfuric add R, if — temperature: 30 °C.
necessary. Extract with 3 quantities, each of 25 mL, of Mobile phase:
methylene chloride R. Combine the methylene chloride extracts — mobile phase A: to 353 m L o f water R add 147 m L of
and wash w ith 2 quantities, each of 8 mL, of solution C and phosphoric add R and mix; dilute 2 m L of the solution to
then once with 10 m L of solution D . Evaporate the organic 1000 m L w ith water R;
layer to dryness at 33 °C, completing the drying procedure — mobile phase B: acetonitrUe Rm
,
using compressed air. Dissolve the residue in 2.0 m L o f the
mobile phase. Time Mobile phase A Mobile phase B
Reference solution (a) Dissolve 7.5 m g of diacerein (min) (per cent V/V) (per cent V/V)
impurity B CRS in tetrahydrofuran R and dilute to 25.0 m L 0 -3 80 20
with the same solvent. Sonicate for n o t more than 30 s. 20*40
3 - 13 8 0 * 60
Dilute 1.0 m L o f the solution to 100.0 m L with solution A.
Dilute 5.0 m L of this solution to 50.0 m L with solution A. 1 3-20 60 40
Mix 5.0 m L o f this solution with 25 m L o f solution A for
10 m in. Add 70 m L o f solution B and adjust to p H 9.5 with
Flow rate 1.2 mlVmin.
dilute sodium hydroxide solution R or dilute sulfuric add R, if
necessary. Perform the extraction as described for the test Detection Spectrophotom eter at 254 nm.
solution. Care should be taken that the time between dissolution Injection 20 jiL.
of diacerdn impurity B in tetrahydrofuran and extraction does not Identification of impurities Use the chromatogram supplied
exceed 30 min. with diacerdn impurity mixture CRS and the chromatogram
Reference solution (b) Dilute 1.0 m L o f reference solution (a) obtained with reference solution (c) to identify the peaks due
to 5.0 m L with the mobile phase. to impurities C and F; use the chromatogram obtained with
Column: reference solution (b) to identify the peaks due to
— sizer. I = 0.125 m, 0 = 4.6 mm; impurities D and E.
— stationary phase: irregular octadecylsifyl silica gel for Relative retention W ith reference to diacerein (retention
chromatography R (5 pm); time = about 13.5 min): impurity D = about 1.1;
— temperature'. 1 6 + 1 °C. impurity E = about 1.15; impurity C = about 1.2;
Mobile phase tetrahydrofuran R, acetonitrUe R, 4 g/L solution of impurity F = about 1.3.
citric add R (8:27.5:64.5 VIV/V). System suitability:
Flow rate 1.0 mlVmin. — resolution: minimum 1.5 between the peaks due to
impurities D and E in the chrom atogram obtained with
Detection Spectrophotom eter at 254 nm .
reference solution (b);
Injection 100 jiL. — signal-to-noise ratio: minimum 100 for the principal peak
Run time 2.5 times the retention time o f impurity B. in the chromatogram obtained with reference solution (a).
Retention time Im purity B = about 11 min. Limits:
System suitability: reference solution (b): — correction factors: for the calculation of content, multiply
— signal-to-noise ratio: minimum 10 for the principal peak. the peak areas of the following impurities by the
Limit: corresponding correction factor: impurity C = 1.4;
— sum of impurities B and H: n o t more than the area o f the impurity D = 1.3; im purity E = 1.3; impurity F = 9.5;
peak due to impurity B in the chromatogram obtained — impurities D, E: for each impurity, not more than 5 times
w ith reference solution (a) (15 ppm). the area of the principal peak in the chromatogram
obtained with reference solution (a) (0.5 per cent);
Related substances
— impurity C: n o t more than twice the area o f the principal
Liquid chrom atography (2.2.29). Cany out the test protected
peak in the chromatogram obtained with reference
from light.
solution (a) (0.2 per cent);
Solvent mixture Mobile phase A, mobile phase B (50:50 V/V). — impurity F: n o t more than 1.5 times the area of the
Test solution Dissolve 0.100 g of the substance to be principal peak in the chrom atogram obtained with
examined in 50 m L of tetrahydrofuran R and dilute to reference solution (a) (0.15 per cent);
100.0 m L with the solvent mixture. — unspecified impurities: for each impurity, not more than the
Reference solution (a) Dilute 1.0 'm l. o f the test solution to area o f the principal peak in the chromatogram obtained
100.0 m L with tetrahydrofuran R. Dilute 1.0 m L of this with reference solution (a) (0.10 per cent);
solution to 10.0 m L w ith the solvent mixture. — total: not more than 20 times the area of the principal
Reference solution (b) In order to prepare impurities D and E peak in the chromatogram obtained with reference
in situ, add 10.0 m L o f 0.01 M sodium hydroxide to 0.100 g of solution (a) (2.0 per cent);
the substance to be examined. Add 40 m L of — disregard limit: 0.5 times the area of the principal peak in
tetrahydrofuran R and dilute to 100.0 m L with the solvent the chromatogram obtained with reference solution (a)
mixture. (0.05 per cent).
2016 Diacerein 1-727
C h ro m iu m
c o 2h
Maximum 10 ppm .
Atomic absorption spectrometry (2.2.23, Method I).
Test solution In a digestion bomb, dissolve 0.25 g o f the
oh o oh
substance to be examined in a mixture o f 2 m L of strong
hydrogen peroxide solution R and 6 mT. of nitric acid R.
C . 4,5-dihydroxy-9,10-dioxo-9,10-dihydroanthracene-2-
Mineralise using a microwave oven with a power-
carboxylic acid (rhein),
incrementing system. Transfer quantitatively to a volumetric
flask with wetter R and dilute to 50.0 m L with water R.
Centrifuge. Dilute 5.0 m L of the clear supernatant to
c o 2h
50.0 m L with water R.
Blank solution Prepare as described for the test solution,
omitting the substance to be examined.
Stock solution Dilute 5.0 m L of chromium standard solution
(100 ppm Cr) R to 50.0 m L with water R. Dilute 5.0 m L of
this solution to 100.0 mT. with water R. Dilute 2.0 mT. o f this
solution to 100.0 m L with a 0.12 per cent VIV solution of D . 5-acetoxy-4-hydroxy-9,10-dioxo-9,10-dihydroanthracene-
dilute nitric acid R. 2-carboxylic acid (monoacetyl rhein isomer A),
Reference solutions Prepare the reference solutions using the
stock solution, diluting with die blank solution.
Source Chromium hollow-cathode lamp using a transmission c o 2h
band preferably o f 0.2 nm.
Wavelength 357.9 nm .
Atomisation device G raphite furnace.
Loss o n d ry in g (2.2.32)
Maximum 0.5 per cent, determined on 1.000 g by drying in
an oven at 105 °C. E. 4-acetoxy-5-hydroxy-9,10-dioxo-9, 10-dihydroanthracene-
S u lfated a s h (2.4.14) 2-carboxylic a d d (monoacetyl rhein isomer B),
Maximum 0.1 per cent, determined on 1.0 g.
ASSAY
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modifications.
Test solution Dissolve 60.0 mg of the substance to be
examined in tetrahydrofuran R and dilute to 50.0 m L with die
same solvent. D ilute 2.0 m L of the solution to 25.0 m L with Y
o
the solvent mixture.
Reference solution Dissolve 60.0 mg o f diacerein CRS in
tetrahydrofuran R and dilute to 50.0 m L with the same
solvent. Dilute 2.0 m L of the solution to 25.0 m L with the
solvent mixture.
Calculate the percentage content o f C i 9H 120 8 taking into
account die assigned content of diacerein CRS.
F. (10S)-3-(acetoxymethyl)-10-(2,3,4,6-tetra-0-acetyl-P-r>-
STORA GE glucopyranosyl)-9-oxo-9,10-dihydroanthracene- 1, 8-diyl
In an airtight container, protected from light. diacetate (heptaacetyl aloin, heptaacetyi barbaloin),
IM P U R IT IE S
Specified impurities B, C, D , E, F, H
Other detectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of
the tests in the m onograph. They are limited by die general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use Y
0
(2034). It is therefore not necessary to identify these
impurities for dem onstration o f compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use): G.
G . 3-(acetoxymethyl)-10-(2,3,4,6-tetra-0-acetyl-P-i>
OH O OH glucopyranosyl) anthracene-1,8 ,9-triyl triacetate,
B. 1,8-dihydroxy-3-(hydroxymethyl)-anthracene-9, 10-dione
(aloe-emodin),
1-728 Diamorphine Hydrochloride 2016
CHROMATOGRAPHIC CONDITIONS
(a) Use a stainless steel colum n (12.5 cm x 4.6 mm) packed
with base-deactivated octylsUyl silica gel for chromatography,
(5 |im) (Lichrospher RP-select B is suitable).
H3C O 0 o ch3
(b) Use isocratic elution and the mobile phase described
T0 T0 below.
(c) Use a flow rate o f 1 m L per minute.
H. 3-(acetoxymethyl)-9j 10-dioxo-9, 1O-dihydroanthracene-
I , 8-diyl diacetate (triacetyl aloe-emodin). (d) Use an am bient colum n temperature.
PhEur
(e) Use a detection wavelength of 283 nm.
(f) Inject 20 pL o f each solution.
(g) Allow die chromatography to proceed for twice the
retention time of the peak due to diamorphine hydrochloride.
Diamorphine Hydrochloride M O B IL E P H A S E
CHARACTERS
A white or almost white, fine or crystalline powder,
practically insoluble in water, freely soluble in
dimethylformamide, slightly soluble in alcohol. It is very
soluble in dilute solutions of the alkali hydroxides.
ID E N T IF IC A T IO N
First identification B.
B. R = C O -C H 2-Cl: 2 -chloro-N-(4-chloro-2-benzoylphenyl)- Second identification A, C, D.
Af-methylacetaroide,
A. Dissolve 50.0 mg in 5 m L of 1 M sodium hydroxide and
D. R = H : [5-chloro-2- dilute to 50.0 m L with water R. D ilute 1.0 m L of this
(methylamino)phenyl] phenylmethanone. solution to 100.0 m L with 0 . 1 M sodium hydroxide. Examined
ch3 between 230 nm and 350 nm (2.2.25), the solution shows an
absorption maximum at 280 nm and a shoulder at 304 nm.
T he specific absorbance at the maximum is 570 to 610.
B. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
diazoxide CRS. Examine the substances prepared as discs
using potassium bromide R.
C. Examine the chromatograms obtained in the test for
C. 3-amino-6-chJoro - 1-methyl-4-phenylquinolin-2(lii)-one, related substances in ultraviolet light at 254 nm.
çh3 T he principal spot in the chromatogram obtained with test
solution (b) is similar in position and size to the principal
spot in the chromatogram obtained with reference
solution (b).
D . Dissolve about 20 mg in a mixture of 5 m L of hydrochloric
add R and 10 m L of water R. Add 0.1 g of zinc powder R.
Boil for 5 min, cool and filter. T o the filtrate add 2 m L of a
1 g/L solution o f sodium nitrite R and mix. Allow to stand for
E. 6-chloro-1-methyl-4-phenylquinazoIin-2( lH)-onc, 1 min and add 1 m L of a 5 g/L solution of
naphthylethylenediamine dihydrochloride R. A red or violet-red
colour develops.
TESTS
Ci A p p e a ra n c e o f so lu tion
Dissolve 0.4 g in 2 m l, of 1 M sodium hydroxide and dilute to
20 m L with water R. The solution is clear (2.2.1) and not
m ore intensely coloured than reference solution Y 7 (2.2.2,
F. 7-chloro-2-m ethoxy-5-phenyl-3/i- 1,4-benzodiazepine. Method II).
A c id ity o r alk alin ity
___ PhEur
T o 0.5 g of the powdered substance to be examined add
30 m L of carbon dioxide-free water R, shake for 2 m in and
filter. T o 10 m L o f the filtrate add 0.2 m L of 0.01 M sodium
★* ★ hydroxide and 0.15 mT. o f methyl red solution R. T he solution
Diazoxide ★ ★ is yellow. N o t more than 0.4 m L of 0.01 M hydrochloric add
★ ★
is required to change the colour o f the indicator to red.
(Ph. Etcr. monograph 0550) *****
R e la te d su b sta n c e s
Examine by thin-layer chromatography (2.2.27), using silica
gel GF2 5 4 R as the coating substance.
Test solution (a) Dissolve 0.1 g of the substance to be
examined in a mixture of 0.5 m L of 1 M sodium hydroxide
and 1 m L o f methanol R and dilute to 5 m L with methanol R.
C8H7a N 20 2S 230.7 364-98-7
Test solution (b) Dilute 1 m L of test solution (a) to 5 m L
A c tio n a n d u se with a mixture of 1 volume of 1 M sodium hydroxide and
Vasodilator; T reatm ent of hypertension. 9 volumes of methanol R.
P re p a r a tio n s Reference solution (a) Dilute 0.5 m L o f test solution (a) to
Diazoxide Injection 100 m L with a mixture of 1 volume of 1 M sodium hydroxide
and 9 volumes of methanol R.
Diazoxide Tablets
Reference solution (b) Dissolve 20 m g of diazoxide CRS in a
PhEur________________________________ mixture o f 0.5 m l, of 1 M sodium hydroxide and 1 m L of
D E F IN IT IO N
methanol R and dilute to 5 m L with methanol R.
Diazoxide contains n o t less than 98.0 per cent and n o t more Apply separately to the plate 5 (jL o f each solution. Develop
than the equivalent o f 101.0 per cent of 7-chloro-3-methyl- over a path o f 15 cm using a mixture o f 7 volumes o f
2ii-l,2,4-benzothiadiazine 1, 1-dioxide, calculated with concentrated ammonia R, 25 volumes of methanol R and
reference to the dried substance.' 68 volumes of chloroform R. Allow the plate to dry in air and
2016 Dibrompropamidine Isetionate 1-731
examine in ultraviolet light at 254 nm. Any spot in the B. Mix 0.1 g w ith 0.5 g o f anhydrous sodium carbonate R,
chrom atogram obtained with test solution (a)3 apart from the ignite and take up the residue with 20 m L of water R. Filter
principal spot3 is not m ore intense than the spot in the and neutralise the filtrate to blue litmus paper R with nitric
chrom atogram obtained with reference solution (a) add R. T he filtrate gives reaction (a) of bromides (2.3.1).
(0.5 per cent).
TESTS
L oss o n d ry in g (2.2.32) p H (2.2.3)
N ot more than 0.5 per cent3 determined on 1.000 g by 5.0 to 6.0.
drying in an oven at 105 °C for 2 h.
Dissolve 0.50 g in carbon dioxide-free water R and dilute to
S u lfa te d a s h (2.4.14) 10 m L with the same solvent.
N ot more than 0.1 per cent3 determined on 1.0 g.
R e la te d su b sta n c e s
A SSA Y Liquid chromatography (2.2.29).
Dissolve 0.200 g with gentle heating in 50 m L of a mixture Solvent mixture anhydrous formic add R, methanol R, ethyl
of 1 volume o f water R and 2 volumes of acetate R (0.01:8:12 VIVIV).
dimethytformamide R. T itrate with 0.1 M sodium hydroxide, Test solution T o 8 m L o f methanol R add 20.0 m g of the
determining the end-point potentiometrically (2.2.20). Carry substance to be examined and dissolve with the aid of an
out a blank titration. ultrasonic bath. Add 11 m L of ethyl acetate R then 10 (iL of
1 m L of 0.1 M sodium hydroxide is equivalent to 23.07 m g of anhydrous formic acid R and mix. Dilute to 20.0 m L with ethyl
C 8H 7GIN 2O 2S. acetate R.
___________________________________________________________ PhEur Reference solution (a) Dilute 1.0 m L of the test solution to
100.0 m L with the solvent mixture. D ilute 1.0 m L of this
solution to 10.0 m L with the solvent mixture.
Reference solution (b) Dissolve 10 mg of dibrompropamidine for
system suitability CRS (containing impurities A and B) in
Dibrompropamidine Isetionate ***** 4 m L of methanol R using an ultrasonic bath. Add 5 m L of
*★ *
(Dibrompropamidine Dnsetionate, ** ethyl acetate R then 5 fiL of anhydrous formic acid R and mix.
Ph Eur monograph 2300) Dilute to 10.0 m L with ethyl acetate R.
Column'.
NH NH — sizer. I = 0.25 m, 0 = 4.6 mm 3
— stationary phase: strong cation-exchange silica gel for
chromatography R (5 pm).
Mobile phase Mix 4 volumes of a 25 g/L solution of
Br Br ammonium formate R in methanol R and 6 volumes of ethyl
9 ,/*\. ^ s o 3h acetate R.
’ 1 HO
Flow rate 1 mL/min.
C j i H ^ N ^ oSj 722 614-87-9 Detection Spectrophotometer at 254 nm.
Injection 40 (iL.
A ctio n a n d u se Run time 1.5 times the retention time of dibrompropamidine.
Antiseptic.
Identification of impurities Use the chromatogram supplied
Ph Eur_______ ____ _____________________________________________ with dibrompropamidine for system suitability CRS and the
chromatogram obtained with reference solution (b) to
D E F IN IT IO N
identify the peaks due to impurities A and B.
3,3 '-D ibrom o-434 '-(propane-133-diylbisoxy) dibenzimidamide
Relative retention W ith reference to dibrompropamidine
bis( 2-hydroxyethanesulfonate).
(retention time = about 20 min): impurity A = about 0.4;
C o n te n t impurity B = about 1.1.
99.0 per cent to 101.0 per cent (dried substance).
System suitability: reference solution (b):
P R O D U C T IO N — peak-to-vaUey ratio: minimum 1.5, where Hp = height
T he production m ethod m ust be evaluated to determine the above the baseline o f the peak due to impurity B and
potential for formation o f alkyl 2-hydroxyethanesulfonates, Hv = height above the baseline of the lowest point o f the
which is particularly likely to occur if the reaction medium curve separating this peak from the peak due to
contains lower alcohols. W here necessary, the production dibrompropamidine.
method is validated to demonstrate that alkyl Limits:
2-hydroxyethanesulfonates are not detectable in the final — impurity A: n o t more than 3 times the area o f the
product. principal peak in the chromatogram obtained with
CHA RACTERS reference solution (a) (0.3 per cent);
— impurity B: n o t more than 5 times the area of the
A p p e a ra n c e
principal peak in the chromatogram obtained with
White or almost white, crystalline powder.
reference solution (a) (0.5 per cent);
S o lubility — unspecified impurities: for each impurity, not more than the
Freely soluble or soluble in water, slightly soluble in ethanol area of the principal peak in the chromatogram obtained
(96 per cent), practically insoluble in methylene chloride. with reference solution (a) (0.1 per cent);
ID E N T IF IC A T IO N — total: not more than 10 times the area of the principal
A. Infrared absorption spectrophotometry (2.2.24). peak in the chromatogram obtained with reference
Comparison dibrompropamidine diisetionate CRS. solution (a) ( 1.0 per cent);
1-732 Dibutyl Phthalate 2016
— disregard, limit. 0.5 times the area of the principal peak in B. Refractive index (2.2.6): 1.490 to 1.495.
the chromatogram obtained with reference solution (a) C. Infrared absorption spectrophotometry (2.2.24).
(0.05 per cent). Comparison dibutyl phthalate CRS.
L o ss on d ry in g ( 2.2.32) D . Thin-layer chromatography (2.2.27).
M axim um 2.0 per cent, determined on 1.000 g by drying in
Test solution Dissolve 50 m g of the substance to be examined
an oven at 105 °C.
in ether R and dilute to 10 m L with the same solvent.
S u lfa te d a s h (2.4.14) Reference solution Dissolve 50 m g of dibutyl phthalate CRS in
M axim um 0.1 per cent, determ ined on 1.0 g.
ether R and dilute to 10 m L with the same solvent.
A SSA Y Plate TLC silica gel GF2 5 4 plate R.
Dissolve 0.250 g in 50 m L o f dimethylformamide R. T itrate Mobile phase heptane R, ether R (30:70 V/V).
w ith 0.1 M tetrabutylammonium hydroxide under a current o f
Application 10 pL.
nitrogen R, determining the end-point
potentiometrically (2.2.20). Development Over a path of 15 cm.
1 m L of 0.1 M tetrabutylammonium hydroxide is equivalent to Drying In air.
36.12 mg of C 2iH 3oBr2N 4OioS 2. Detection Examine in ultraviolet light at 254 nm.
IM P U R IT IE S Results T he principal spot in the chromatogram obtained with
Specified impurities A, B. the test solution is similar in position and size to the principal
spot in the chromatogram obtained with the reference
nh x solution.
E. T o about 0.1 m L add 0.25 m L o f sulfuric add R and
50 m g of resordnol R. H eat in a water-bath for 5 min. Allow
to cool. Add 10 m L of water R and 1 m L o f strong spdium
R Br hydroxide solution R The solution becomes yellow of
brownish-yellow and shows a green fluorescence.
A. R = Br, X = O: 3-brom o-4-[3-(2-bromo-4- TESTS
carbamimidoyiphenoxy)propoxy]benzamide, Appearance
B. R = H, X = N H : 3-bromo-4-[3- T h e substance to be examined is clear (2.2.1) and not more
(4-carbamimidoylphenoxy)propoxy] benzimidamide. intensely coloured than reference solution Y 6 (2.2.2,
PhEur Method II).
Acidity
Dissolve 20.0 g in 50 m l. of ethanol (96per cent) R previously
neutralised to phenolphthaldn solution R l. Add 0.2 m L of
★ ★ phenolphthalein solution R l. N o t more than 0.50 m L of 0.1 M
Dibutyl Phthalate ★ ★ sodium hydroxide is required to change the colour of the
(Ph. Eur. monograph 0762) ***** indicator.
Related substances
Gas chromatography (2.2.28).
Internal standard solution Dissolve 60 mg of bibenzyl R in
Irgecdon 1 |iL. mp
Run time 3 times the retention time o f dibutyl phthalate. A bout 59 °C.
Elution order Bibenzylj dibutyl phthalate. ID E N T IF IC A T IO N
Retention time Dibutyl phthalate = about 12 min. Infrared absorption spectrophotometry (2.2.24).
System suitability: Comparison 2,4-dichlorobenzyl alcohol CRS.
— resolution: minimum 12 between the peaks due to bibenzyl TESTS
and dibutyl phthalate in the chromatogram obtained with
Related substances
the reference solution;
Liquid chromatography (2.2.29).
— in the chromatogram obtained with test solution (a), there
is no peak with the same retention time as the internal Solvent mixture acetonitrUe R l, water R (50:50 V/V).
standard. Buffer solution Dissolve 0.68 g of potassium dihydrogen
Limit: phosphate R in 900 m L of water R, adjust to pH 3.0 with
— total: calculate the ratio (R) of the area o f the peak due to phosphoric add R and dilute to 1000.0 m L with water R.
dibutyl phthalate to the area of the peak due to the Test solution (a) Dissolve 0.100 g o f the substance to be
internal standard from the chromatogram obtained with examined in 10.0 m L of acetonitrUe R l and dilute to 50.0 m L
the reference solution; from the chromatogram obtained with the solvent mixture.
with test solution (b), calculate the ratio of the sum of the Test solution (b) Dilute 5.0 m L of test solution (a) to
areas of any peaks, apart from the principal peak and the 50.0 m L with the solvent mixture.
peak due to the internal standard, to the area of the peak Reference solution (a) Dilute 1.0 m L of test solution (a) to
due to the internal standard: this ratio is not greater than 100.0 m L with the solvent mixture. Dilute 1.0 m L of this
R (1.0 per cent). solution to 10.0 m L with the solvent mixture.
Water (2.5.12) Reference solution (b) Dissolve 20.0 m g of 2¡4-dichlorobenzyl
Maximum 0.2 per cent, determined on 10.00 g. alcohol impurity A CRS and 20.0 m g of 2, 4-dichlorobenzyl
S u lfa te d a sh (.2.4.14) alcohol impurity C CRS in 100.0 m L of acetomtrüe R l. Dilute
M aximum 0.1 per cent, determined on 1.0 g. 1.0 m L o f the solution to 100.0 m L with the solvent mixture.
ASSAY Reference solution (c) Dissolve 0.100 g of 2,4-dichlorobenzyl
Introduce 0.750 g into a 250 m L borosilicate glass flask. alcohol CRS in 10.0 m L of acetomtrüe R l and dilute to
Add 25.0 m L of 0.5 M alcoholic potassium hydroxide and a few 50.0 m L with the solvent mixture. Dilute 5.0 m L of the
glass beads. H eat in a water-bath under a reflux condenser solution to 50.0 m L with the solvent mixture.
for 1 h. Add 1 m i. o f phenolphthalein solution R1 and titrate Column:
immediately with 0.5 M hydrochloric acid until the colour — size. I — 0.15 m , 0 = 4.6 mm;
changes from red to colourless. Carry out a blank titration. — stationary phase, end-capped octadecylsüyl silica gel for
Calculate the volume o f potassium hydroxide used in the chromatography R (5 |im);
saponification. — temperature. 30 °C.
1 m L o f 0.5 M alcoholic potassium hydroxide is equivalent to Mobile phase.
69.59 m g of CKSH22O 4. — mobile phase A: methanol R2, acetonitrUe R l, buffer solution
(20:30:50 V/V/V);
STORA GE — mobile phase B: acetonitrUe R l ;
In an airtight container.
__________________________________________________________ PhEur
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0-7 100 0
7 - 17 100*20 0 * 80
2,4-Dichlorobenzyl Alcohol ***** 17 -3 0 20 80
(Ph. Eur. monograph 2410) *
Flow rate 1.2 mL/min.
Detection Spectrophotometer at 214 nm.
Irtjection 10 jiL o f test solution (a) and reference solutions (a)
and (b).
CvHeClaO 177.0 1777-82-8 Relative retention W ith reference to 2,4-dichlorobenzyl alcohol
PhEur__________________________________________________________ (retention time = about 7 min): impurity C = about 0.87;
impurity A = about 0.91.
D E F IN IT IO N
(2,4-EHchlorophenyl)methanol. System suitability, reference solution (b):
— peak-to-vaUey ratio: minimum 4, where Hp = height above
C o n te n t the baseline of the peak due to impurity C and
98.0 p er cent to 102.0 per cent (anhydrous substance). Hv = height above the baseline of the lowest point o f the
CHARACTERS curve separating this peak from the peak due to
A p p e a ra n c e impurity A.
W hite or almost white, crystalline powder. Calculation of percentage contents:
S o lu b ility — for each impurity, use the concentration of
Very slightly soluble in water, very soluble in ethanol 2,4-dichlorobenzyl alcohol in reference solution (a).
(96 per cent).
1-734 Dichloromethane 2016
Limits:
— unspecified impurities: for each impurity, maximum
0.10 per cent;
— total: maximum 0 .3 per cent;
— reporting threshold: 0.05 p er cen t F. 2,4-dichlorobenzaldehyde,
W a te r (2.5.32)
M axim um 0.2 per cent, determined on 0.500 g using the
evaporation technique:
— temperature: 120 °C .
S u lfa te d a s h (2.4.14) G. 1,1 /-(oxydimethylene)bis(2,4-dichlorobenzene).
M axim um 0.1 per cent, determined on 1.0 g.
PhEur
A SSAY
Liquid chrom atography (2.2.29) as described in the test for
related substances w ith the following modification.
Injection T est solution (b) and reference solution (c).
Calculate the percentage content of CyHôC^O taking into
Dich loromethane / *
account the assigned content of 2,4-dichlorobenzyl (Methylene Chloride, Ph. Eur. monograph 0932) *
alcohol CRS.
CH 2C12 84.9 75-09-2
IM P U R IT IE S
Other detectable impurities (the following substances would, if A ctio n a n d u se
present at a sufficient level, be detected by one or other of Excipient.
the tests in the monograph. They are limited by the general
PhEur__________ :_______________________________________________
acceptance criterion fo r other/unspecified impurities and/or
by the general m onograph Substances for pharmaceutical use D E F IN IT IO N
(2034). It is therefore not necessary to identify these D ichloromethane.
impurities for dem onstration of compliance. See also 5.10. It may contain maximum 2.0 per cent V/V of anhydrous
Control of impurities in substances for pharmaceutical use): A, B, ethanol and/or maximum 0.03 per cent V/V of 2-methylbut-
C, D, E, F, G. 2-ene as stabiliser.
CHARACTERS
A p p e a ra n c e
Clear, colourless, volatile liquid.
S o lu b ility
Sparingly soluble in water, m isdble with ethanol
A. (2,5-dichlorophenyl)methanol,
(96 per cent).
ci ID E N T IF IC A T IO N
First identification B, C.
Second identification A, D, E.
A. Relative density (see Tests).
B. Refractive index (see Tests).
B, (2, 6-dichlorophenyl) methanol,
C. Infrared absorption spectrophotometry (2.2.24).
Preparation Films.
Comparison methylene chloride CRS.
D . H eat 2 m L with 2 g o f potassium hydroxide R and 20 m L
ci of ethanol (96 per cent) R under a reflux condenser for
30 min. Allow to cool. Add 15 m L of dilute sulfuric acid R
C. (3,4-dichlorophenyl)methanol, and filter. T o 1 m L of the filtrate add 1 m L of a 15 g/L
solution o f chromotropic acid, sodium salt R, 2 m L of water R
o and 8 m L o f sulfuric acid R. A violet colour is produced.
E. 2 m L o f the filtrate obtained in identification test D gives
reaction (a) of chlorides (2.3.1).
TESTS
A p p e a ra n c e
D. 2,4-dichlorobenzyl acetate, It is clear (2.2.1) and colourless (2.2.2, Method IT).
A cid ity
T o 50 m L o f methanol R previously neutralised to 0.1 m L of
bromothymol blue solution R l, add 50 g of the substance to be
examined. N o t more than 0.15 m L o f 0.1 M sodium hydroxide
is required to change the colour o f the indicator to blue.
E. 2,4-dichlorobenzoic ad d , R elativ e d e n sity (2.2.5)
1.320 to 1.332.
2016 Dichloromethane 1-735
R e fra c tiv e in d ex (2.2.6) — sum of impurities other than ethanol and 2-methyBmt-2-ene:
1.423 to 1.425. maximum 0.1 per cent F/F;
E th a n o l, 2 -m eth y Ib u t-2 -en e a n d v o latile im p u ritie s — reporting threshold: 50 ppm F/F; the reporting threshold
Gas chromatography (2.2.28). does n o t apply to impurity A.
B. Carry out the m ethod for thin-layer chromatography, are about 25 minutes for diclofenac and about 12 minutes
Appendix HI A, using the following solutions in methanol for diclofenac impurity A.
(1) 5.0% w/v o f the substance being examined. MOBILE PHASE
(2) 5.0% w/v of diclofenac diethylamine BPCRS. 34 volumes of a mixture o f equal volumes of a 0.1% w/v
CHROMATOGRAPHIC CONDITIONS solution o f orthophosphoric acid and a 0.16% w/v solution of
sodium dihydrogen orthophosphate adjusted to p H 2.5 and
(a) Use a silica gel precoated plate (Macherey Nagel SIL
66 volumes of methanol
G -25 H R or silica gel 6OF254 H P T L C plates are suitable).
(b) Use the mobile phase as described below. SYSTEM SUITABILITY
(c) Apply 2 (jL o f each solution. T he test is not valid unless, in the chromatogram obtained
with solution (3), the resolution factor betw een the peaks
(d) Develop the plate to 15 cm.
corresponding to diclofenac and diclofenac im purity A is at
(e) After removal of the plate, dry it in a stream of warm air least 6.5.
for 10 minutes. Spray with nmhydrin solution and heat at 110°
LIMITS
for 15 minutes.
In the chromatogram obtained with solution ( 1):
MOBILE PHASE
the area of any secondary peak is n o t greater than the area of
1 volume of hydrochloric acid, 1 volume of water, 6 volumes of
the principal peak in the chromatogram obtained with
glacial acetic acid and 11 volumes of ethyl acetate.
solution (2 ) (0 .2 %);
SYSTEM SUITABILITY the sum o f the areas of any secondary peaks is not greater than
T h e test is not valid unless the chromatogram obtained with 2.5 times the area of the principal peak in the chromatogram
solution (2) shows two clearly separated spots. obtained with solution (2) (0.5%).
CONFIRMATION Disregard any peak with an area less than 0.25 times the area
T he two principal spots in the chromatogram obtained with of the principal peak in the chromatogram obtained with
solution ( 1) are similar in position, colour and size to the solution (2) (0.05%).
corresponding spots in the chromatogram obtained with Loss o n d ry in g
solution ( 2). W hen dried at a pressure not exceeding 1 kPa for 24 hours,
TESTS loses not more than 0.5% of its weight. Use 1 g.
Acidity or alkalinity S u lfa te d a sh
pH of a 1% w/v solution in ethanol (10%), 6.4 to 8.4, N o t more than 0.1%, Appendix IX A, M ethod IL Use 1 g.
A ppendix V L. A SSAY
Clarity and colour o f solution Dissolve 0.5 g in 30 m L of anhydrous acetic acid and carry
A 5% w/v solution in methanol is dear, Appendix IV A. T h e out M ethod I for rum-aqueous titration, Appendix V IE A,
absorbance o f the solution measured at 440 nm is n o t greater determining the end point potentiometrically. E ach m L o f
than 0.05, Appendix II B. 0.1m perchloric add KS is equivalent to 36.93 mg of
Heavy metals C 1 3 H 2 2 C I2 N 2 O 2 .
2 g complies with limit test C for heavy metals, Appendix VH. STO R A G E
U se 2 m L of lead standard solution (10 ppm Pb) to prepare the Diclofenac Diethylamine should be kept in an airtight
standard (10 ppm). container and protected from light.
Related substances IM P U R IT IE S
Carry out the m ethod for liquid chromatography,
T h e impurities limited by the requirem ents of this
Appendix HI D , using the following solutions in the mobile
monograph include those listed under Diclofenac Sodium.
phase.
( 1) 0 . 10% w/v o f the substance being examined.
(2) D ilute 2 volumes of solution (1) to 100 volumes and
dilute 1 volume of this solution to 10 volumes.
(3) Dissolve 1 m g of diclofenac impurity A BPCRS add 1 m L
Diclofenac Potassium *****
*+ +*
o f solution (1) and dilute to 200 mL. (Ph. Eur. monograph 1508) *
CHROMATOGRAPHIC CONDITIONS
(a) Use a stainless steel column (25 cm x 4.6 m m ) packed
with end-capped octylsilyl silica gd for chromatography (5 Jim)
(end-capped Zorbax C 8 is suitable).
(b) U se isocratic elution and the mobile phase described
below.
(c) Use a flow rate of 1 m L per minute.
(d) Use an am bient column temperature.
C 14H 10CI2KNO 2 334.2 15307-81-0
(e) Use a detection wavelength of 254 nm.
(f) Inject 20 jiL of each solution. A c tio n a n d u se
(g) Allow the chromatography to proceed for 1.5 times the Cyclo-oxygenase inhibitor, analgesic; anti-inflammatory.
retention time o f diclofenac. PhEur__________________________________________________________
Inject 20 (iL o f solution (3). W hen the chromatograms are
D E F IN IT IO N
recorded under the prescribed conditions, the retention times
Potassium [2-[(2,6-dichlorophenyl)amino]phenyl] acetate.
1-738 Diclofenac Potassium 2016
Solubility Column:
— size: I = 0.25 m, 0 = 4.6 mm;
Sparingly soluble in water, freely soluble in methanol, soluble
— stationary phase: end-capped octadecylsUyl silica gelfor
in ethanol (96 per cent), slighdy soluble in acetone.
chromatography R (5 pm).
IDENTIFICATION Mobile phase Mix 34 volumes of a solution containing 0.5 g/L
First identification: A, D. of phosphoric acid R and 0.8 g/L of sodium dihydrogen
Second identification B, C, D phosphate R, previously adjusted to p H 2.5 with phosphoric
A. Infrared absorption spectrophotometry (2.2.24). add R, and 66 volumes o f methanol R.
Comparison diclofenac potassium CRS. Flow rate 1.0 mL/min.
B. Thin-layer chromatography (2.2.27). Detection Spectrophotom eter at 254 nm.
Test solution Dissolve 25 mg o f the substance to be examined Injection 20 pL.
in methanol R and dilute to 5 m L with the same solvent. Run time 1.6 times the retention time of diclofenac.
Reference solution (a) Dissolve 25 mg of diclofenac Identification of impurities Use the chromatogram supplied
potassium CRS in methanol R and dilute to 5 m L with the with diclofenac for system suitability CRS and the
same solvent. chromatogram obtained with reference solution (b) to
Reference solution (b) Dissolve 10 mg o f indometacin R in identify the peaks due to impurities A and F.
reference solution (a) and dilute to 2 m L with the same Relative retendon W ith reference to diclofenac
solution. (retention time = about 25 min): impurity A = about 0.4;
Plate TLC silica gel GF2 5 4 plate R. impurity F = about 0.8.
Mobile phase concentrated ammonia R, methanol R, ethyl System suitability, reference solution (b):
acetate R (10:10:80 VIVIV). — resolution: minimum 4.0 between the peaks due to
Application 5 pL. im purity F and diclofenac.
A ctio n a n d u se
Cyclo-oxygenase inhibitor; analgesiq anti-inflammatory.
A. 1-(2 j 6-dichlorophenyl)-1,3-dihydro-2/i-indol-2-onej P re p a ra tio n s
Prolonged-release Diclofenac Capsules
Gastro-resistant Diclofenac Tablets
Prolonged-release Diclofenac Tablets
PhEur__________________________________________________________
D E F IN IT IO N
Sodium [2- [(2,6-dichlorophenyl) amino] phenyl] acetate.
5 0 -6 0 100 0
W a te r (;2.5.12) H ea v y m e ta ls
M axim um 0.2 per cent, determ ined on 10.0 g. 12 m L o f a 10.0% w/v solution complies w ith limit test A for
S u lfa te d a sh (2.4.14) heavy metals, Appendix VII. Use lead standard solution
M axim um 0.1 per cent, determ ined on 1.0 g. (1 ppm Pb) to prepare the standard (10 ppm ).
S u lfa te
A SSA Y
0.6 g complies with the limit test for sulfates, Appendix VII
Introduce 0.750 g into a 250 m L borosilicate glass flask.
(250 ppm ).
A dd 25.0 m L o f 0.5 M alcoholic potassium hydroxide and a few
glass beads. Boil in a water-bath under a reflux condenser for L oss o n d ry in g
1 h. Add 1 m L of phenolphthalein solution R l and titrate W hen dried at 60° for 3 hours, loses not m ore than 0.1% of
immediately with 0.5 M hydrochloric acid. Carry out a blank its w eight Use 1 g.
titration. Calculate the volume of 0.5 M alcoholic potassium A SSA Y
hydroxide used in the saponification. Carry out M ethod I for non-aqueous titration,
1 m L of 0.5 M alcoholic potassium hydroxide is equivalent to A ppendix VHI A, using 0.4 g and 1-naphtholbenzein solution
55.56 mg of C 12H 14O 4. as indicator. Each m L o f 0.1m perchloric acid KS is equivalent
ST O R A G E to 21.13 m g o f C 11H 17N O 3.
In an airtight container. STORAGE
___________________________________________________________PhEur Diethylamine Salicylate should be protected from lig h t
It should n o t be allowed to come into contact with iron or
iron salts.
Diethylamine Salicylate
COOH
Diethylcarbamazine Citrate *****
** +*
(PL Eur. monograph 0271) *
o
ho c o 2h
V CHa , HOaC^JX^COjH
Cn H 17N 0 3 211.3 4419-92-5
H3C" ch3
A c tio n a n d u se
C ounter-irritant. C 16H 29N30 8 391.4 1642-54-2
P re p a r a tio n
Diethylamine Salicylate Cream A c tio n a n d u se
Antihelminthic.
D E F IN IT IO N P r e p a r a tio n
Diethylamine Salicylate contains n o t less than 99.0% and n o t Diethylcarbamazine Tablets
m ore than 101 .0 % o f C 11H 17N O 3.
PhEur__________________________________________________________
C H A R A C T E R IS T IC S
W hite or alm ost white crystals. D E F IN IT IO N
Very soluble in water, freely soluble in ethanol (96%). NjiV-Diethyl-4-methylpiperazine-1 -carboxamide dihydrogen
2-hydroxypropane- 1,2,3-tricarboxylate.
ID E N T IF IC A T IO N
C o n te n t
A. T h e infrared absorption spectrum, Appendix II A, is
98.0 per cent to 102.0 per cent (dried substance).
concordant with the reference spectrum o f diethylamine
salicylate (RS 099). CHARACTERS
B. T o 0.2 g add 5 m L of 1m sodium hydroxide, heat to boiling A p p e a ra n c e
point, cool and acidify with 2 m hydrochloric add; a white W hite or alm ost white, crystalline powder, slighdy
precipitate is produced. T he melting point of the precipitate, hygroscopic.
after recrystallisation from water and drying at 105°, is about S o lu b ility
160°, Appendix V A. Very soluble in water, soluble in ethanol (96 per cent),
TESTS practically insoluble in acetone.
A cid ity mp: about 138 °C, with decomposition.
Dissolve 2 g in 25 m L o f water and titrate with 0.1m sodium ID E N T IF IC A T IO N
hydroxide VS using phenol red solution as indicator. N ot more First identification A , C
than 0.2 m L o f 0.1m sodium hydroxide KS is required to
Second identification B, C.
change the colour o f the solution.
A. Infrared absorption spectrophotometry (2.2.24).
C la rity a n d c o lo u r o f so lu tio n
A 50% w/v solution is dear, A ppendix IV A, and not more
Comparison diethylcarbamazine citrate CRS.
intensely coloured than reference solution BYS) Appendix IV B, B. Examine the chromatograms obtained in the test for
M ethod II. impurities A and B.
M e ltin g p o in t Results T h e principal spot in the chromatogram obtained with
100° to 102°, A ppendix V A. the test solution is similar in position, colour and size to the
2016 Diethylcarbamazine Citrate 1-747
principal spot in the chromatogram obtained with reference Reference solution (d) Dissolve 5.0 mg o f diethykarbamazme
solution (a). citrate CRS in solution A and dilute to 50.0 m L with
C. Dissolve 0.1 g in 5 m L o f water R. T he solution gives the solution A.
reaction of citrates (2.3.1). Column:
— size: I = 0.15 m, 0 = 3.9 mm;
TESTS
— stationary phase: end-capped octadecylsüyl silica gel for
S o lu tio n S
chromatography R (5 pm).
Shake 2.5 g with water R until dissolved and dilute to 25 m L
with the same solvent. Mobile phase M ix 100 volumes o f methanol R2 and
900 volumes of a 10 g/L solution of potassium dihydrogen
A p p e a ra n c e o f so lu tio n phosphate R.
Solution S is n o t more opalescent than reference
suspension II (2.2.1) and n o t more intensely coloured than
Flow rate 0.8 mL/min.
reference solution BY6 (2.2.2, Method II). Detection Spectrophotometer at 220 nm.
Im p u ritie s A a n d B Injection 20 pL o f test solution (a) and reference solutions (a),
Thin-layer chromatography (2.2.27). (b) and (c).
Test solution Dissolve 0.5 g of the substance to be examined Run time Twice the retention time of diethylcarbamazine.
in methanol R and dilute to 10 m L with the same solvent. Identification of impurities Use the chromatogram obtained
Reference solution (a) Dissolve 0.1 g o f diethykarbamazme with reference solution (b) to identify the peak due to the
citrate CRS in methanol R and dilute to 2.0 m L with the same citrate.
solvent. Relative retention W ith reference to diethylcarbamazine
Reference solution (b) Dissolve 10 mg of methylpiperaztne R (retention time = about 7 min): citrate = about 0.2;
(impurity A) in methanol R and dilute to 100 m L w ith the degradation product = about 1.6.
same solvent. System suitability: reference solution (c):
Reference solution (c) Dissolve 10 mg o f dtmethylpiperazine R — resolution: minimum 5 between the -peaks due to
(impurity B) in methanol R and dilute to 100 m L w ith the diethylcarbamazine and the degradation product.
same solvent. Limits:
— unspecified impurities: for each impurity, not more than the
Plate TLC silica gel plate R.
area of the principal peak in the chromatogram obtained
Mobile phase concentrated ammonia R, methyl ethyl ketone R, with reference solution (a) (0.10 per cent);
methanol R (5:30:65 V/VIV). — total: n o t more than 5 times the area o f the principal peak
Application 10 pL. in the chromatogram obtained with reference solution (a)
Development Over 2/3 of the plate. (0.5 per cent);
Drying A t 100-105 °C. — disregard limit. 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (a)
Detection Expose to iodine vapour for 30 min.
(0.05 per cent); disregard the peak due to the citrate.
Retardation factors Impurity A = about 0.2;
impurity B = about 0.5. H eav y m e ta ls (2.4.8)
M aximum 20 ppm.
Limits:
— impurity A: any spot due to impurity A is not more 12 m L of solution S complies with test A. Prepare the
intense than the corresponding spot in the chromatogram reference solution using 10 m L o f lead standard solution
obtained with reference solution (b) (0.2 per cent); (2 ppm Pb) R.
— impurity B: any spot due to impurity B is not m ore L oss o n d ry in g (2.2.32)
intense than the corresponding spot in the chromatogram Maximum 0.5 per cent, determined on 1.000 g by drying
obtained with reference solution (c) (0.2 per cent). in vacuo at 60 °C for 4 h.
R elated su b sta n c e s S u lfated a s h (2.4.14)
Liquid chromatography (2.2.29). Maximum 0.1 per cent, determined on 1.0 g.
Solution A Dissolve 31.2 g o f potassium dihydrogen phosphate R ASSAY
in water R and dilute to 1000 m L with the same solvent. Liquid chromatography (2.2.29) as described in the test for
Test solution (a) Suspend 0.30 g of the substance to be related substances with the following modification.
examined in solution A and dilute to 100 m L with Injection 20 pL o f test solution (b) and reference solution (d).
solution A. Filter or centrifuge and use the clear filtrate or
Calculate the percentage content o f C 16H 29N30 8 from the
supernatant.
declared content of diethylcarbamazine citrate CRS.
Test solution (b) Dissolve 10.0 mg of the substance to be
examined in solution A and dilute to 100.0 m L with STORAGE
solution A. In an airtight container.
Reference solution (a) Dilute 1.0 m L of test solution (a) to IM P U R IT IE S
100.0 m L with solution A. Dilute 1.0 m L of this solution to Specified impurities A, B
10.0 m L with solution A.
Reference solution (b) Dissolve 10 mg o f citric add R in
solution A and dilute to 10 m L with solution A.
H3C 'Nv'" ^
Reference solution (c) T o 3 m L o f test solution (a) add 0.5 m L
of strong hydrogen peroxide solution R and maintain at 80 °C A. R = H: 1-methylpiperazine,
for 3 h. Dilute to 100 m L with solution A.
B. R = C H 3: 1,4-dimethylpiperazine.
PhEur
1-748 Diethylene Glycol Monoethyl Ether 2016
— stationary phase:
Diethylene Glycol Monoethyl Ether *****
pdy(oyanopropyl) (7) (phenyl) (7) ( methyl) (86)sUoxane R
(Ph. Eur. monograph 1198) * (film thickness 1 pm).
Carrier gas nitrogen for chromatography R o r helium for
chromatography R.
Flow rate 2.0 mlVmin.
Q H 14O 3 134.2 111-90-0 Split ratio 1:80.
Temperature:
A c tio n a n d u se
Excipient.
Time Temperature
PftEir __________________________________________________________ (min) CO
Column 0 -1 120
D E F IN IT IO N
2-(2-Ethoxyethoxy)ethanol, produced by condensation of 1-10 120 -» 225
ethylene oxide and alcohol, followed by distillation. 10-12 225
CHARACTERS Injection port 275
A p p e a ra n c e Detector 250
Clear, colourless, hygroscopic liquid.
S o lu b ility
Miscible with water, w ith acetone and with alcohol, m isable Detection Flam e ionisation.
in certain proportions with vegetable oils, n o t miscible with Injection 0.5 pL.
mineral oils. Relative retentions W ith reference to diethylene glycol
R elativ e d e n sity monoethyl ether (retention time = about 4 m in): ethylene
A bout 0.991. glycol monom ethyl ether = about 0.4; ethylene glycol
monoethyl ether = about 0.5; ethylene glycol = about 0.55;
ID E N T IF IC A T IO N
diethylene glycol = about 1. 1.
A. Refractive index (2.2.6): 1.426 to 1.428.
System suitability:
B. Infrared absorption spectrophotometry (2.2.24). — resolution: minim um 3.0 between the peaks due to
Comparison Ph. Eur. reference spectrum of diethylene glycol ethylene glycol monoethyl ether and to ethylene glycol in
monoethyl ether. the chromatogram obtained with reference solution (b),
TESTS — signal-to-noise ratio: minim um 3.0 for the peak due to
A cid v alu e (2.5.1) ethylene glycol monomethyl ether in th e chrom atogram
M axim um 0.1. obtained with reference solution (a),
Mix 30.0 m L with 30 m L o f alcohol R previously neutralised Limits (take into account the impurity/internal standard peak
with 0.1 M potassium hydroxide using phenolpkthalein area ratio):
solution R as indicator. T itrate with 0.01 M alcoholic potassium — ethylene glycol monomethyl ether, not m ore than the area of
hydroxide. the corresponding peak in the chrom atogram obtained
with reference solution (a) (50 ppm ),
P e ro x id e v a lu e (2.5.5)
— ethylene glycol monoethyl ether, n o t m ore th an the area of
M aximum 8.0, determ ined on 2.00 g.
the corresponding peak in the chrom atogram obtained
R e la te d su b s ta n c e s with reference solution (a) (160 ppm ),
Gas chromatography (2.2.28). — ethylene glycol: not more than the area o f the
Internal standard solution Dilute 1.00 g of decane R to corresponding peak in the chrom atogram obtained with
100.0 m L w ith methanol R. reference solution (a) (620 ppm ),
Test solution T o 5.00 g of the substance to be examined, add — diethylene glycol: not m ore than the area o f the
0.1 m L of the internal standard solution and dilute to corresponding peak in the chrom atogram obtained with
10.0 m L with methanol R. reference solution (a) (250 ppm),
— total: no t more than the area of the principal peak in the
Reference solution (a) D ilute 25.0 mg o f ethylene glycol
chrom atogram obtained with reference solution (c)
monomethyl ether R, 80.0 m g of ethylene glycol monoethyl
(0.2 per cent).
ether R, 0.310 g of ethylene glycol R and 0.125 g o f diethylene
glycol R to 100.0 m L w ith methanol R. T o 1.0 m L of this E th y le n e ox id e
solution add 0.1 m L o f the internal standard solution and H ead-space gas chromatography (2.2.28).
dilute to 10.0 m L with methanol R. Test solution T o 1.00 g o f the substance to be examined in a
Reference solution (b) D ilute 25.0 mg of ethylene glycol vial, add 50 pL of water R.
monoethyl ether R and 25.0 m g of ethylene glycol R to Reference solution T o 1.00 g o f the substance to be examined
100.0 m L w ith methanol R. Dilute 1.0 m l. o f this solution to in a vial, add 50 pL of ethylene oxide solution R4 and close
5.0 m L with methanol R. tighdy.
Reference solution (c) D ilute 1.00 g of the substance to be Column:
examined to 100.0 m L with methanol R. T o 1.0 m L o f this — material: fused silica,
solution add 0.1 m l. o f the internal standard solution and — size. I = 30 m , 0 = 0.32 mm,
dilute to 10.0 m L with methanol R. — stationary phase:
Column: poly(cyanopropyl) (7) (phenyl) (7) ( methyl) (86)sUoxane R
— material: fused silica, (film thickness 1 pm).
— size. I = 30 m , 0 = 0.32 mm, Carrier gas helium for chromatography R.
2016 Diethylene Glycol Palmitostearate 1-749