Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

Dedications
―To All Students of Pharmaceutical Sciences‖

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

Table of Contents
INTRODUCTION PHARMACEUTICALS QUALITY MANAGEMENT _______________________________________ 7

Basic concepts ____________________________________________________________________________________________ 8

Introduction of pharmaceutical industry ______________________________________________________________________ 8

Testing _________________________________________________________________________________________________ 11

Quality Management System _______________________________________________________________________________ 13

Pharmaceutical Quality Management System _________________________________________________________________ 16

Continual Improvement Of Process Performance And Product Quality _____________________________________________ 18

Quality Assurance ________________________________________________________________________________________ 21

Quality Assurance System (QAS) ____________________________________________________________________________ 22

Standard Operating Procedures (SOPs): ______________________________________________________________________ 22

Good Manufacturing Practices ______________________________________________________________________________ 23

Current Good Manufacturing Practices _______________________________________________________________________ 23

Current Good Manufacturing Practice for Finished Pharmaceuticals _______________________________________________ 24

Validation_______________________________________________________________________________________________ 28

QALITY CONTROL OF SOLID DOSAGE FORMS ___________________________________________________________ 33

Dosage Forms: ___________________________________________________________________________________________ 34

Hardness (BP) Or Breaking Force (USP) _______________________________________________________________________ 35

Thickness Diameter _______________________________________________________________________________________ 36

Friability ________________________________________________________________________________________________ 36

Disintegration ___________________________________________________________________________________________ 39

Weight Variation _________________________________________________________________________________________ 45

Content uniformity _______________________________________________________________________________________ 48

DISSOLUTION TEST (USP) __________________________________________________________________________________ 49

Assay of active Ingredient _________________________________________________________________________________ 53

QUALITY CONTROL OF SYRUPS, ELIXIRS, AND DISPERSE SYSTEM __________________________________ 56

Viscosity its determination and application in the Quality Control of Pharmaceuticals ________________________________ 58

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

ASSAY & CONTENT UNIFORMITY ____________________________________________________________________________ 65

1. FOR SYRUPS: __________________________________________________________________________________________ 65

2. FOR ELIXIR ____________________________________________________________________________________________ 66

3. FOR EMULSION ________________________________________________________________________________________ 68

4. FOR SUSPENSION ______________________________________________________________________________________ 69

Weight per ml ___________________________________________________________________________________________ 70

QUALITY CONTROL OF SUPPOSITORIES _________________________________________________________________ 71

Visual Examination:_______________________________________________________________________________________ 72

Weight Variation Test _____________________________________________________________________________________ 72

Melting Range Test _______________________________________________________________________________________ 72

Breaking Test ____________________________________________________________________________________________ 73

Disintegration Test (Liquefaction Test or Sogtening Test) ________________________________________________________ 73

Content Uniformity (EP) ___________________________________________________________________________________ 76

Dissolution Test: _________________________________________________________________________________________ 76

QUALITY CONTROL OF STERILE PRODUCTS (PARENTERALS) _______________________________________ 77

STERILE PRODUCTS _______________________________________________________________________________________ 78

Sterility Test _____________________________________________________________________________________________ 78

Leaker’s test _____________________________________________________________________________________________ 87

Clarity test ______________________________________________________________________________________________ 88

Visible Test (visual assessment of parenterals) ________________________________________________________________ 90

Pyrogen Test ____________________________________________________________________________________________ 91

TEST FOR PYROGENS ______________________________________________________________________________________ 92

Assay for active Ingredient _________________________________________________________________________________ 95

Safety Test (Remington) ___________________________________________________________________________________ 95

BIOLOGICAL ASSAYS ________________________________________________________________________________________ 96

ASSAYS _________________________________________________________________________________________________ 97

BIOASSAYS: _____________________________________________________________________________________________ 97

Bioassay of Antibiotics ____________________________________________________________________________________ 99

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

Bioassay of Insulin Injection _______________________________________________________________________________ 100

The Rabbit Blood Sugar Method ___________________________________________________________________________ 100

Dextrose Determination __________________________________________________________________________________ 102

Assay of prepared digitalis ________________________________________________________________________________ 103

Assay of Vitamin D. ______________________________________________________________________________________ 104

ALCOHOL DETERMINATION ______________________________________________________________________________106

Methods To Determine Alcohol ____________________________________________________________________________ 107

Method I (Distillation Method) ____________________________________________________________________________ 107

METHOD II (GAS LIQUID CHROMATOGRAPHY)________________________________________________________________ 109

ALKALOIDAL DRUG ASSAY ________________________________________________________________________________111

Alkaloids ______________________________________________________________________________________________ 112

ALKALOIDAL DRUG ASSAY ________________________________________________________________________________ 112

Examples: ______________________________________________________________________________________________ 113

QUALITY ASSURANCE OF VACCINES _____________________________________________________________________117

Tests of general Applications ______________________________________________________________________________ 120

MISCELLANEOUS DETERMINATIONS AND TESTS _____________________________________________________123

DETERMINATION OF WEIGHT/mL __________________________________________________________________________ 124

Water Content Determination _____________________________________________________________________________ 124

LOSS ON DRYING ________________________________________________________________________________________ 127

IDENTIFICATION TESTS ___________________________________________________________________________________ 127

IN VIVO IDENTIFICATION TESTS: ___________________________________________________________________________ 128

IDENTIFICATION TESTS IN VITRO ___________________________________________________________________________ 128

EVALUATION OF OINTMENT ______________________________________________________________________________ 130

Microbial content test: ___________________________________________________________________________________ 133

Metal particle test: ______________________________________________________________________________________ 133

Sterility test: ___________________________________________________________________________________________ 133

Potency/content uniformity test: __________________________________________________________________________ 134

Viscosity: ______________________________________________________________________________________________ 134

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

Leakage test ____________________________________________________________________________________________ 135

Homogenicity: __________________________________________________________________________________________ 135

ASH CONTENT DETERMINATION:___________________________________________________________________________ 135

ALKALINITY OF GLASS ____________________________________________________________________________________ 136

STATISTICAL INTERPRETATION OF QUALITY CONTROL CHARTS DURING MANUFACTURING

PROCESSES __________________________________________________________________________________________________138

Quality ________________________________________________________________________________________________ 139

Quality control__________________________________________________________________________________________ 139

Quality Assurance _______________________________________________________________________________________ 139

Statistical quality control (SQC) ____________________________________________________________________________ 139

Control Charts __________________________________________________________________________________________ 141

Types of Control Charts __________________________________________________________________________________ 142

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

INTRODUCTION
PHARMACEUTICALS
QUALITY MANAGEMENT

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Adnan’s Pharmaceutical Quality Management
Basic concepts
The pharmaceutical environment today is changing
quickly due to globalization, increased competition,
cost constraints, demands for efficiency, and
development of international regulation, supply
chain complexity, and product/process complexity.
In this fast-changing environment, the people and
companies that learn to adapt will prosper to
manufacture & deliver consistently zero-defect
products to the patients.
The quality, efficacy and safety attributes of
products must be ensured so that the consumer
health is not compromised.
In the drug industry at large, quality management is
usually defined as the aspect of management
function that determines and implements the
―quality policy‖ i.e. the overall intention and
direction of an organization regarding quality, as
formally expressed and authorized by top
management. The basic elements of quality
management are:
An appropriate infrastructure or ―quality system‖,
encompassing the organizational structure,
procedures, processes and resources;
Systematic actions necessary to ensure adequate
confidence that a product (or service) will satisfy
given requirements for quality. The totality of these
actions is termed quality assurance.
Within an organization, quality assurance serves as
a management tool. In contractual situations, quality
assurance also serves to generate confidence in the
supplier. The concepts of quality assurance, GMP
and quality control are interrelated aspects of
quality management. They are described here in
order to emphasize their relationship and their
fundamental importance to the production and
control of pharmaceutical products.
Quality control
Quality control is a process employed to ensure a
certain level of quality in a product or service.
Adnan Sarwar Chaudhary
It can be defined as: “A system for ensuring the
maintenance of proper standards in manufactured
goods, especially by periodic random inspection of
the product”.
OR
“Quality control refers to a procedure or a set of
steps taken during the manufacturing of a product
or service to ensure that it meets requirements and
that the product/service is reproducible”.
Quality Assurance
Quality assurance is a wide ranging concept that
covers all matters that individually or collectively
influence the quality of a drug. It starts from
purchase to post market surveillance.
QA = QC + GMP’s
Pharmaceutical QC:
“It is mainly concerned with the analytical
measurement processes and focuses on technical
aspects of testing of drugs at all stages.”
Introduction of
pharmaceutical industry
Quality Control is one of the key departments in
any Pharma company. After R&D large number of
people works in the QC department. A
chemist/Pharmacist executing a qualitative analysis
seeks to identify the substances in the sample. A
quantitative analysis is an attempt to determine the
quantity or concentration of a specific substance in
the sample.
Quality Control department in
Pharma Industry
Quality Control had two different divisions
1. Wet Analysis
2. Instrumental Analysis
The Quality will be checked in three different
stages.
1) Raw material analysis
2) In Process Sample analysis
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3) Finished Product analysis 16) TLC (Thin Layer Chromatography)

17) Water Bath
Once the raw material enters the factory premises 18) Dissolution apparatus
and before going to the Stores department the
Quality of the material will be checked by QC Qualification required to work in QC department
department. If the quality is as per the guidelines
then the QC department approves the raw material. M.SC Chemistry/B.Pharm/M.Pharm/Pharm-D/ M
This is called Raw material analysis. The concerned Phil/Ph.D
QC chemist will perform the basic duties and the
Group leader or Manager approves. Career Path
The In Process Analysis will be done while the
Entry Level
Product (Chemical or Formulation) is being
1. Trainee
Prepared/Manufactured
2. Chemist
Finished Product analysis will be done after the
3. Sr.Chemist
Product/material is manufactured.
4. Officer
The different Instruments used in QC department
Middle Level
are:
1. Executive/Sr.Executive
1) UPLC
2. Asst/Dy.Manager
2) HPLC
3. Manager
3) GC
4. Sr.Manager
4) UV
5) FTIR Sr.Level
6) AAS 1. AGM/DGM
7) XRD 2. G.M
8) Particle size analyzer 3. Sr.GM
9) KF Tritator
10) MV Tritator
11) Melting Point apparatus
12) Muffle Furnace
13) Polarimeter
14) Friability Apparatus
15) Disentrigation Apparatus

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Quality Assurance Quality Control

Definition QA is a set of activities for ensuring QC is a set of activities for ensuring
quality in the processes by which quality in products. The activities focus
products are developed. on identifying defects in the actual
products produced.
Focus on QA aims to prevent defects with a QC aims to identify (and correct) defects
focus on the process used to make the in the finished product. Quality control,
product. It is a proactive quality therefore, is a reactive process.
process.
Goal The goal of QA is to improve The goal of QC is to identify defects
development and test processes so that after a product is developed and before
defects do not arise when the product it's released.
is being developed.
How Establish a good quality management Finding & eliminating sources of quality
system and the assessment of its problems through tools & equipment so
adequacy. Periodic conformance audits that customer's requirements are
of the operations of the system. continually met.
What Prevention of quality problems The activities or techniques used to
through planned and systematic achieve and maintain the product
activities including documentation. quality, process and service.
Responsibility Everyone on the team involved in Quality control is usually the
developing the product is responsible responsibility of a specific team that
for quality assurance. tests the product for defects.
Verification is an example of QA Validation/Software Testing is an
example of QC
Example
Statistical Statistical Tools & Techniques can be When statistical tools & techniques are
Technique applied in both QA & QC. When they applied to finished products (process
are applied to processes (process outputs), they are called as Statistical
inputs & operational parameters), they Quality Control (SQC) & comes under
are called Statistical Process Control QC.
(SPC); & it becomes the part of QA.
As a tool QA is a managerial tool QC is a corrective tool

Standard
Something established as a measure or model to
which other similar things/materials should
conforms.
Further two types:
a. Reference Standard
b. Working Standard
Reference Standard
Drug substance of highest purity which is
reasonably attainable, specifically prepared by
independent synthesis or by further purification of
existing production material and shown to be
authentic by extensive set of analytical tests.
Working Standard
Drug substance of established quality and purity as shown by
comparison to the reference standard material
and used for routine quality.
Assay (Standardization)
The word assay comes from the French word assai,
which means "trial".
It is type of analysis for the determination of
amount of particular constituent in a mixture or
biological and pharmacological potency of a drug. It
also refers to the measurement of active ingredient
in a dosage form. For example, an assay may be
done of a vaccine to determine its potency.

OR

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Determination of activity, potency and strength
of substances either on absolute basis or in
comparison with that of standard.
Biological Assay:
Bioassays are typically conducted to measure the
effects of a substance on a living organism and
are
essential in the development of new drugs. Both
are procedures by which the potency
(pharmacology) or
the nature of a substance is estimated by
studying its effects on living matter.
Chemical Assay:
It deals with the study of the chemical
composition of substances. More broadly, it may
be considered the corpus of all techniques
whereby any exact chemical information is
obtained.
It has further two branches:
Qualitative assay:
Qualitative assay is the determination of those
elements and compounds that are present in a
sample of unknown material.
Quantitative assay:
Quantitative assay is the determination of the
amount by weight of each element or compound
present.
Testing
A quality assurance program should include testing
of finished compounded preparations. It is
important for the compounder to have a basic
understanding of pharmaceutical analysis to ensure
that valid results are obtained when tests are being
conducted, whether they are done in-house or
outsourced. While it is not practical to test every
compounded preparation, it is incumbent on the
compounder to know
(1) The importance of testing in the overall quality
program in the compounding facility,
(2) When to test,
(3) What to test,
Adnan Sarwar Chaudhary
(4) What method(s) to use,
(5) How to interpret the results,
(6) The limits of the test, and
(7) What to do if the preparations listed do not meet
specifications.
Investigative and corrective action should extend to
other preparations that may have been associated
with the specific failure or discrepancy.
The goal in testing is to produce results as
accurately, efficiently, and quickly as possible. Any
testing method used should have accuracy, speed,
reproducibility, and specificity. No single testing
method is suited for all drugs. There are a number
of factors that determine the validity and reliability
of results.
Compounding facilities have two options when
testing is required. Some testing methods can easily
be performed in-house, but some may need to be
outsourced to a contract laboratory.
Relatively basic testing methods that can be
conducted in-house with proper training and a
modest investment in instrumentation include
weight and volumetric measurements, pH,
density/specific gravity, refractive index, and UV
and visible spectroscopy (see Weights and Balances
41, Volumetric Apparatus 31, Prescription Balances
and Volumetric Apparatus 1176, pH 791, Specific
Gravity 841, Refractive Index 831, and
Spectrophotometry and Light-Scattering 851).
Testing methods often outsourced to a contract
laboratory include chromatography (high-pressure
liquid chromatography (HPLC) and gas
chromatography (GC), see Chromatography 621),
mass spectroscopy (MS) (see Mass Spectrometry
736), hyphenated methods (HPLC-MS and GC-
MS), UV and visible spectroscopy (see
Spectrophotometry and Light-Scattering 851), and
other sophisticated methods.
If testing is done in-house, appropriate equipment
must be obtained, verified either by the
manufacturer or by the compounder upon purchase,
maintained, calibrated, and used properly. If testing
is outsourced, the compounder needs to determine
what to outsource and how to select a laboratory,
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and should develop ongoing relationships with the
laboratories chosen. Contract laboratories should
follow USP general chapter standards, as
appropriate.
Selection of a Testing Method
One general consideration in testing method
selection is the type of information that is needed,
such as quantitative (potency, concentration), semi
quantitative (where a tolerance level is involved, as
in endotoxin levels), or qualitative
(presence/absence type of testing, including
substance identification, sterility). Another
consideration involves the physical and chemical
characteristics of the analyte, including solubility,
partition coefficient, dissociation constant (pKa),
volatility, binding, and the quantity present.
The degree of quantitative measurement and
specificity must be considered in the validation
process. The typical analytical characteristics used
in method validation include accuracy, precision,
specificity, detection limit, quantitation limit,
linearity, range, and ruggedness (see Validation of
Compendial Procedures 1225). Generally, the
greater the level of accuracy, precision, or
specificity required the more sophisticated and
expensive the testing methods needed. The methods
used are also governed by the types of
instrumentation available and the standards
available for comparison.
Pharmaceutical analysis decisions include not only
method selection but also administrative and
economic factors, obtaining a representative
sample, storage/shipping of the sample, sample
preparation for analysis, the actual analysis, data
acquisition, data treatment, and interpretation.
Factors Involved in Method
Selection
The testing method selected depends upon a number
of factors, including sample requirements, sample
handling/preparation/purification requirements, type
of data needed, and levels of specificity and
accuracy required.
Adnan Sarwar Chaudhary
Sampling Requirements
Prior to collecting samples for testing, the following
factors should be considered: the number of
samples needed, appropriate methods of obtaining
representative samples, the physical state of the
samples (solid, liquid, or gas), the type of container
required for collection and storage, and possible
shipping requirements or restrictions. Storage
requirements for samples must be specified, such as
type of container, temperature, humidity, and light
protection (see General Notices and Requirements).
The effect of any substances in the formulation that
may interfere or alter the results must be known
beforehand. When sending a preparation to a
contract laboratory, the compounder should provide
the complete formulation so the laboratory can
quickly determine if there may be any interfering
substances.
Controlled drug substances, dangerous or hazardous
chemicals, flammable or caustic substances, and
refrigerated or frozen preparations require special
handling during shipping.
Data Interpretation Requirements
The collection of raw data from the testing process
must be completed accurately. One must ensure that
appropriate and valid descriptive statistics are used
to analyze the data, and that the operating
parameters of the analytical instruments are well
established. Reference values, if available, should
be provided with the analytical results. A
description of the analytical controls used by the
laboratory is important for documentation, as well
as the source of reference standards used to
establish standard curves.
Personnel Requirements and
Considerations
If testing is done in-house, personnel involved in
this activity must be appropriately trained and
evaluated with documentation of the training and
evaluation. If testing is outsourced, the compounder
must be assured of the credentials, proper training,
and continuing competency activities of the
personnel in the contract laboratory.
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It is preferable that the contract laboratory be

registered with the Food and Drug Administration
(FDA). Also, it may be advantageous if the contract
laboratory performs testing for pharmaceutical
companies. Immunoassay

Testing Methods
(EMIT)
Testing methods can be generally divided into
physical testing methods, methods that interact with
electromagnetic radiation, conduct metric
techniques, immunoassay methods, separation
Separation Techniques
techniques, and others.

Classification of Analytical (HPLC)

Methods s Chromatography (GC)
-Layer Chromatography (TLC)
Physical Testing Procedures

Others

Testing
r change Nonspecific methods include melting, freezing and
boiling point.

Quality Management System

Quality of medicines means meeting the required
specifications. Quality management in
pharmaceutical industries, is an important
Interaction of Electromagnetic Radiation subject
because the drugs / or pharmaceutical products
are directly delivered to the customers body
system, thus identity, purity safety and
Fluorescence/Phosphorescence spectroscopy ultimately appropriate quality of product are
strongly essential. ICH (CH Stands for International
-ray spectroscopy Conference on Harmonization of Technical Requirements for
Registration of Pharmaceuticals for Human Use”. ICH is a
spectroscopy joint initiative involving both regulators and research-based
industry representatives of the EU, Japan and the US in
scientific and technical discussions of the testing procedures
required to assess and ensure the safety, quality and efficacy
of medicines. Harmonization process – founded April 1990)
Conductance Methods Guidelines are established with a view to bring

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uniformity in the products worldwide. It helps in
export and import of drug products worldwide.
Maintaining quality in the products is a complex
process and needs to take into account various
guidelines like GMP, GLP and many more. There
is a Quality assurance department in all the
Pharma industries whose job is to look if all the
required guidelines are being followed in the
industries or not. Quality auditing is the process
through which they check internally or
externally and ensure everything is running
right.
In the present scenario the context of Quality has
emerged as an important factor. People are wise
enough to choose things that assure to fulfill their
demands. If we precisely define Quality
it means meeting the specifications that are
summarized keeping in mind the demand of today‘s
fast changing world. If we talk of Pharmaceutical
Industry, quality becomes an unavoidable thing.
Quality management in pharmaceutical industries,
is an important subject because the drugs / or
pharmaceutical products are directly delivered to
the customers body system, thus identity, purity
safety and ultimately appropriate quality of product
are strongly essential. There are numerous
guidelines worldwide that has made some
sort of rules and specifications which must be
followed by every pharmaceutical industry. To
maintain quality in pharmaceutical products,
Quality Management System is followed.
Internationally harmonized guidance ICH Q10 (ICH
Q Documents Q1 Stability Q2 Analytical Validation Q3
Impurities Q4 Pharmacopoeias Q5 Quality
Biotechnological Products Q6 Specifications Q7 Good
Manufacturing Practice Q8 Pharmaceutical Development Q9
Quality Risk Management Q10 Pharmaceutical Quality
Systems) governs the concept of current
pharmaceutical quality management system for
Registration of Pharmaceuticals for Human Use and
USFDA and in final phases.
Quality Management System
(QMS)
Quality management is defined as the aspect of
management function that determines and
Adnan Sarwar Chaudhary
implements the “quality policy”, i.e. the overall
intention and direction of an organization
regarding quality, as formally expressed and
authorized by top management.
The basic elements of quality
management are
An appropriate infrastructure or ―quality system‖,
encompassing the organizational structure,
procedures, processes and resources
Systematic actions necessary to ensure that a
product (or service) will satisfy given requirements
for quality. The totality of these actions is termed
―quality assurance‖.
Responsibilities of the
Pharmaceutical Quality Unit
1. To establish the quality system
2. To audit compliance to the quality system
3. To establish procedures and specifications
4. To establish manufacturing controls
5. To perform laboratory tests or examinations
6. To review and approve or reject all things
cGMP
7. To ensure investigation of nonconformance
8. To keep management informed
9. To describe responsibilities in writing
10. To remain independent.
Elements of Quality Management
System
of
A quality management system
typically consists of four facets
a) Quality planning: Process of translating quality
policy into processes, procedures, and instructions
to achieve measurable objectives and requirements
b) Quality assurance: Planned and methodical
activities executed as part of a quality system to
provide confidence that process, product, or service
requirements for quality are being satisfied
c) Quality control: Act of monitoring, appraising,
and correcting a process, product, or service to
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Adnan’s Pharmaceutical Quality Management
ensure requirements for quality are being satisfied
d) Quality improvement: Process of analyzing
performance and taking methodical, systemic
actions to improve it.
International Conference on
Harmonization
ICH is a joint initiative involving both Regulators
and Research based industry initiatives of the
Europe, Japan and US for the scientific and
technical discussions of the testing procedures;
required to assess and ensure the Safety, Quality
and Efficacy of the medicines. ICH stands for
―International Conference on Harmonization‖ of
Technical Requirements for Registration of
Pharmaceuticals for Human use.
ICH Q10
ICH Q10 describes one comprehensive model for
an effective pharmaceutical quality system that is
based on International Organization for
Standardization (ISO). Quality concepts, includes
applicable good manufacturing practice (GMP)
regulations, and complements. Implementation of
ICH Q10 throughout the product lifecycle should
facilitate innovation and continual improvement and
strengthen the link between pharmaceutical
development and manufacturing activities.
Aim
ICH was established in 1990, as a joint
regulatory/industry project to improve, through
harmonization, the efficiency of the process, for
developing and registering new medicinal products
in Europe, Japan and US.
Purpose of ICH
The basic purpose of ICH are
a) To monitor, update and increase the international
harmonization of Technical Requirements.
b) To ensure Safety, Efficacy and Quality of
medicines that must be developed and registered in
the most efficient and cost effective manner.
c) To promote and protect public health from an
international perspective.
Adnan Sarwar Chaudhary
d) To prevent unnecessary duplication of clinical
trials in humans.
e) To minimize the use of animal testing without
compromising the safety and effectiveness.
f) To improve the efficiency of Global Drug
Development.
Need of ICH
The guideline helped in achieving harmonization in
the quality of products worldwide for the export of
medicines without any interruption on the world
level.
Quality Management System in
Testing Laboratories
Professionals of testing laboratories have shown
increasing interest in understanding the QMS and
attaining accreditation status for their services since
the introduction of international standards for the
quality management system (QMS). Thus Quality
assurance therefore is defined as the process or the
end of the process which confirms for the integrity
of a product to meet the standard for the intended
use. Quality assurance is an obligation
automatically imposed on the manufacturer of any
product to ensure that it meets the needs of the end-
user in the measures intended for use. For the end-
user, the benchmark of quality is perfection they
cannot allow less than 100%.
Elements of the Quality
Management System
The laboratory is a complex system, involving
many steps of activity and many people. The
complexity of the system requires that many
processes and procedures be performed properly.
Therefore, the QMS model, which looks at the
entire system, is very important for achieving good
laboratory performance. The QMS is defined as a
‗management system to direct and control an
organization with regard to quality. The QMS
covers the laboratory activities, including drug
sampling, analysis and reporting. The QMS consists
of documentation of the laboratory policy and
objectives, system procedures and instructions for
assuring the quality of its results to meet safety and
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regulatory requirements and to satisfy the needs of ICH Q10 Objectives

the customers.
Implementation of the Q10 model should result in
achievement of three main objectives that
Pharmaceutical Quality complement or enhance regional GMP
requirements.
Management System
1. achieve Product Realization
It is applicable to drug products, including 2. Establish and Maintain a State of Control
biotechnology and biological products, throughout 3. Facilitate Continual Improvement
the product lifecycle the systems supporting the
development and manufacture of pharmaceutical MANAGEMENT
drug substances. It includes: RESPONSIBILITY
 Pharmaceutical Development Leadership is essential to establish and maintain a
o Drug substance development company-wide commitment to quality and for
the performance of the pharmaceutical quality
o Formulation development (including
system.
container/closure system)
o Manufacture of investigational Management Commitment
products  Senior management has the ultimate
o Delivery system development (where responsibility to ensure an effective
relevant) pharmaceutical quality system is in place to
o Manufacturing process development achieve the quality objectives, and that roles,
and scale-up responsibilities, and authorities are defined,
o Analytical method development communicated, and implemented
 Technology Transfer throughout the company.
o New product transfers during  Management should:
development through manufacturing 1. Participate in the design,
o Transfers within or between implementation, monitoring, and
manufacturing and testing sites for maintenance of an effective
marketed products pharmaceutical quality system.
 Commercial Manufacturing 2. Demonstrate strong and visible
o Acquisition and control of materials support for the pharmaceutical
o Provision of facilities, utilities, and quality system and ensure its
equipment implementation throughout their
o Production (including packaging and organization.
labeling) 3. Ensure a timely and effective
o Quality control and assurance communication and escalation
o Release process exists to raise quality issues
o Storage to the appropriate levels of
o Distribution (excluding wholesaler management.
activities) 4. Define individual and collective
 Product Discontinuation roles, responsibilities, authorities,
o Retention of documentation and interrelationships of all
o Sample retention organizational units related to the
o Continued product assessment and pharmaceutical quality system.
reporting Ensure these interactions are
communicated and understood at all

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levels of the organization. An
independent quality unit/structure
with authority to fulfill certain
pharmaceutical quality system
responsibilities is required by
regional regulations.
5. Conduct management reviews of
process performance and product
quality and of the pharmaceutical
quality system.
6. Advocate continual improvement.
7. Commit appropriate resources.
Quality Policy
1. Senior management should establish a
quality policy that describes the overall
intentions and direction of the company
related to quality.
2. The quality policy should include an
expectation to comply with applicable
regulatory requirements and should facilitate
continual improvement of the
pharmaceutical
quality system.
3. The quality policy should be communicated
to and understood by personnel at all
levels in the company.
4. The quality policy should be reviewed
periodically for continuing effectiveness.
Quality Planning
1. Senior management should ensure the
quality objectives to implement the quality
policy are defined and communicated.
2. Quality objectives should be supported by
all relevant levels of the company.
3. Quality objectives should align with the
company‘s strategies and be consistent with
the quality policy.
4. Management should provide the appropriate
resources and training to achieve the quality
objectives.
5. Performance indicators that measure
progress against quality objectives should be
established, monitored, communicated
regularly, and acted upon as appropriate.
Adnan Sarwar Chaudhary
Resource Management
1. Management should determine and provide
adequate and appropriate resources (human,
financial, materials, facilities, and
equipment) to implement and maintain the
pharmaceutical quality system and
continually improve its effectiveness.
2. Management should ensure that resources
are appropriately applied to a specific
product, process, or site.
Internal Communication
1. Management should ensure appropriate
communication processes are established
and implemented within the organization.
2. Communications processes should ensure
the flow of appropriate information between
all levels of the company.
3. Communication processes should ensure the
appropriate and timely escalation of certain
product quality and pharmaceutical quality
system issues.
Management Review
1. Senior management should be responsible
for pharmaceutical quality system
governance through management review to
ensure its continuing suitability and
effectiveness.
2. Management should assess the conclusions
of periodic reviews of process performance
and product quality and of the
pharmaceutical quality system.
Management of Outsourced Activities and
Purchased Materials
The pharmaceutical quality system, including the
management responsibilities described in this
section, extends to the control and review of any
outsourced activities and quality of purchased
materials. The pharmaceutical company is
ultimately responsible to ensure processes are in
place to assure the control of outsourced activities
and quality of purchased materials. These processes
should incorporate quality risk management and
include:
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1. Assessing prior to outsourcing operations or
selecting material suppliers, the suitability
and competence of the other party to carry
out the activity or provide the material using
a defined supply chain (e.g., audits, material
evaluations, qualification).
2. Defining the responsibilities and
communication processes for quality-related
activities of the involved parties. For
outsourced activities, this should be included
in a written agreement between the contract
giver and contract acceptor.
3. Monitoring and review of the performance
of the contract acceptor or the quality of
the material from the provider, and the
identification and implementation of any
essential improvements.
4. Monitoring incoming ingredients and
materials to ensure they are from approved
sources using the agreed supply chain.
Management of Change in Product Ownership
When product ownership changes (e.g., through
acquisitions), management should consider the
complexity of this and ensure:
1. The ongoing responsibilities are defined for
each company involved
2. The essential information is transferred
Continual Improvement Of
Process Performance And
Product Quality
Lifecycle Stage Goals
The goals of each product lifecycle stage are
described below.
 Pharmaceutical Development
The goal of pharmaceutical development activities
is to design a product and its manufacturing
process to consistently deliver the intended
performance and meet the needs of patients and
healthcare professionals, and regulatory authorities
and internal customers‘ requirements.
Adnan Sarwar Chaudhary
 Technology Transfer
The goal of technology transfer activities is to
transfer product and process knowledge between
development and manufacturing, and within or
between manufacturing sites to achieve product
realization. This knowledge forms the basis for the
manufacturing process, control strategy,
process validation approach, and ongoing continual
improvement.
 Commercial Manufacturing
The goals of manufacturing activities include
achieving product realization, establishing and
maintaining a state of control, and facilitating
continual improvement. The pharmaceutical
quality system should assure that the desired
product quality is routinely met, suitable process
performance is achieved, the set of controls are
appropriate, improvement opportunities are
identified and evaluated, and the body of knowledge
is continually expanded.
 Product Discontinuation
The goal of product discontinuation activities is to
manage the terminal stage of the product
lifecycle effectively. For product discontinuation, a
predefined approach should be used to manage
activities such as retention of documentation and
samples and continued product assessment (e.g.,
complaint handling and stability) and reporting in
accordance with regulatory requirements.
Pharmaceutical Quality System Elements
These four elements are:
1. Process performance and product quality
monitoring system
2. Corrective action and preventive action
(CAPA) system
3. Change management system
4. Management review of process
performance and product quality
1. Process Performance and Product
Quality Monitoring System
An effective monitoring system provides assurance
of the continued capability of processes and
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controls to produce a product of desired quality and Process and Monitoring A well- Once
product during defined manufacturin
to identify areas for continual improvement. The
knowledge scale-up system g
process performance and product quality generated activities can for process ceases,
monitoring system should: and process provide a performance monitoring
and preliminary and such as
product indication of product stability
a. Use quality risk management to establish the
monitoring process quality testing
control strategy. This can include conducted performance monitoring should
parameters and attributes related to drug throughout and the should be continue to
development successful applied to completion of
substance and drug product materials and can be integration assure the
components, facility and equipment used to into performance studies.
operating conditions, in-process controls, establish a manufacturi within a Appropriate
control ng. state of action on
finished product specifications, and the strategy for Knowledge control and marketed
associated methods and frequency of manufacturi obtained to product
ng. during identify should
monitoring and control. The control strategy transfer and improvemen continue to
should facilitate timely scale-up t be executed
feedback/feedforward and appropriate activities can areas. according to
be useful in regional
corrective action and preventive action. further regulations.
b. Provide the tools for measurement and developing
analysis of parameters and attributes the control
strategy.
identified in the control strategy (e.g., data
management and statistical tools).
2. Corrective Action and Preventive
c. Analyze parameters and attributes identified
Action (CAPA) System
in the control strategy to verify continued
operation within a state of control. The pharmaceutical company should have a system
d. Identify sources of variation affecting for implementing corrective actions and preventive
process performance and product quality for actions resulting from the investigation of
potential continual improvement activities to complaints, product rejections, nonconformances,
reduce or control variation. recalls, deviations, audits, regulatory inspections
e. Include feedback on product quality from and findings, and trends from process performance
both internal and external sources (e.g., and product quality monitoring. A structured
complaints, product rejections, approach to the investigation process should be
nonconformances, recalls, deviations, audits used with the objective of determining the root
and regulatory inspections, and findings). cause. The level of effort, formality, and
f. Provide knowledge to enhance process documentation of the investigation should be
understanding, enrich the design space commensurate with the level of risk, in line with
(where established), and enable innovative ICH Q9. CAPA methodology should result in
approaches to process validation. product and process improvements and enhanced
product and process understanding.
Application of Process Performance and Product
Quality Monitoring System throughout the Application of Corrective Action and Preventive
Product Lifecycle Action System Throughout the Product Lifecycle
Pharmaceutic Technology Commercial Product Pharmaceuti Technolog Commercial Product
al Transfer Manufacturi Discontinuati cal y Manufacturi Discontinuati
Development ng on Development Transfer ng on

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

Product or CAPA can CAPA CAPA should criteria for a proposed change should be
process be used as should be continue
set.
variability is an effective used, after the
system for and the product is (d) After implementation, an evaluation of the
explored.
feedback, effectiveness discontinued. change should be undertaken to confirm the
CAPA feedforwar of The
methodology d, the actions impact on change objectives were achieved and that
is useful and should be product there was no deleterious impact on product
where continual evaluated. remaining on quality.
corrective improveme the
actions and nt. market should
be Application of Change Management System
preventive Throughout the Product Lifecycle
considered, as
actions are well as Pharmaceuti Technology Commercial Product
incorporated other products cal Transfer Manufacturi Discontinuati
into the that Development ng on
iterative might be Change is an The change A formal Any changes
design and affected. inherent managemen change after
development part of the t system management product
development should system discontinuatio
process.
process and provide should be in n
should be managemen place for should go
3. Change Management System documented; t and commercial through an
the documentati manufacturin appropriate
The change management system should include the formality of on of g. change
the change adjustments Oversight by management
following, as appropriate for the stage of the
management made to the system.
lifecycle: process the process quality unit
should be during should
(a) Quality risk management should be utilized consistent technology provide
with the transfer assurance of
to evaluate proposed changes. The level of
stage of activities. appropriate
effort and formality of the evaluation should pharmaceuti science
be commensurate with the level of risk. cal and risk-
development based
(b) Proposed changes should be evaluated . assessments.
relative to the marketing authorization, 4. Management Review of Process
including design space, where established, Performance and Product Quality
and/or current product and process
understanding. There should be an Management review should provide assurance that
assessment to determine whether a change to process performance and product quality are
the regulatory filing is required under managed over the lifecycle. Depending on the size
regional requirements. As stated in ICH Q8, and complexity of the company, management
working within the design space is not review can be a series of reviews at various levels
considered a change (from a regulatory of management and should include a timely and
filing perspective). However, from a effective communication and escalation process to
pharmaceutical quality system standpoint, raise appropriate quality issues to senior levels of
all changes should be evaluated by a management for review.
company‘s change management system.
(a) The management review system should include:
(c) Proposed changes should be evaluated by
expert teams contributing the appropriate (1) The results of regulatory inspections and
expertise and knowledge from relevant areas findings, audits and other assessments, and
(e.g., Pharmaceutical Development, commitments made to regulatory authorities
Manufacturing, Quality, Regulatory Affairs, (2) Periodic quality reviews, that can
and Medical) to ensure the change is include:
technically justified. Prospective evaluation

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Adnan’s Pharmaceutical Quality Management
(i) Measures of customer satisfaction
such as product quality complaints and
recalls
(ii) Conclusions of process
performance and product quality monitoring
(iii) The effectiveness of process and
product changes including those arising
from corrective action and preventive
actions
(3) Any follow-up actions from previous
management reviews
(b) The management review system should identify
appropriate actions, such as:
(1) Improvements to manufacturing
processes and products
(2) Provision, training, and/or realignment
of resources.
Quality Assurance
Principle.―Quality assurance‖ is a wide-ranging
concept covering all matters that individually or
collectively influence the quality of a product. It is
the totality of the arrangements made with the
object of ensuring that pharmaceutical products are
of the quality required for their intended use.
Quality assurance therefore incorporates GMP and
other factors, including those outside the
scope of this guide such as product design and
development.
The system of quality assurance appropriate to the
manufacture of pharmaceutical products
should ensure that:
(a) Pharmaceutical products are designed and
developed in a way that takes account of the
requirements of GMP and other associated codes
such as those of good laboratory practice
(GLP) and good clinical practice (GCP);
(b) Production and control operations are clearly
specified in a written form and GMP
requirements are adopted;
Adnan Sarwar Chaudhary
(c) Managerial responsibilities are clearly specified
in job descriptions;
(d) Arrangements are made for the manufacture,
supply and use of the correct starting and
packaging materials;
(e) All necessary controls on starting materials,
intermediate products, and bulk products and
other in-process controls, calibrations, and
validations are carried out;
(f) The finished product is correctly processed and
checked, according to the defined procedures;
(g) Pharmaceutical products are not sold or supplied
before the authorized persons have certified that
each production batch has been produced and
controlled in accordance with the requirements of
the marketing authorization and any other
regulations relevant to the production, control and
release of pharmaceutical products;
(h) Satisfactory arrangements exist to ensure, as far
as possible, that the pharmaceutical products
are stored by the manufacturer, distributed, and
subsequently handled so that quality is
maintained throughout their shelf-life;
(i) There is a procedure for self-inspection and/or
quality audit that regularly appraises the
effectiveness and applicability of the quality
assurance system;
(j) Deviations are reported, investigated and
recorded;
(k) There is a system for approving changes that
may have an impact on product quality;
(l) Regular evaluations of the quality of
pharmaceutical products should be conducted with
the objective of verifying the consistency of the
process and ensuring its continuous improvement.
The manufacturer must assume responsibility for
the quality of the pharmaceutical products
to ensure that they are fit for their intended use,
comply with the requirements of the marketing
authorization and do not place patients at risk due to
inadequate safety, quality or efficacy. The
attainment of this quality objective is the
responsibility of senior management and requires
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Adnan’s Pharmaceutical Quality Management
the participation and commitment of staff in many
different departments and at all levels within the
company, the company‘s suppliers, and the
distributors. To achieve the quality objective
reliably there must be a comprehensively designed
and correctly implemented system of quality
assurance incorporating GMP and quality control. It
should be fully documented and its
effectiveness monitored. All parts of the quality
assurance system should be adequately staffed
with competent personnel, and should have suitable
and sufficient premises, equipment, and
facilities.
Quality Assurance System
(QAS)
Formal Structures or techniques to make sure
products and services consistently meet the standard
required by the customer; quality systems may be
validated either within your organization, or by
external auditors or by both.
The basic steps followed to implement system for
an organization are:
Types of SOPs:
 Develop the system
 Document it
 Inform, instruct and train the staff to use it.
Features Of Quality Assurance System
 Pharmaceutical products are designed and
developed in a way that takes account of the
requirements of GMP and other associated
codes such as those of good laboratory
practice (GLP) and good clinical practice
(GCP).
 All necessary controls on starting materials,
intermediate products, and bulk products and
other in process controls, calibrations and
validations are carried out.
 The finished products is correctly processed
and checked according to the defined
procedures.
 Satisfactory arrangements exist to ensure, that
the pharmaceutical products are stored by the
Adnan Sarwar Chaudhary
manufacturer, distributed and subsequently
handled.
 There is a procedure for self-inspection.
 Deviation are reported, investigated and
recorded
 There is a system for approving changes that
may have an impact on product quality.
 Regular evaluations of the quality of
pharmaceutical products should be conducted.
Standard Operating
Procedures (SOPs):
"A Standard Operating Procedure is a document
which describes the regularly recurring operations
relevant to the quality of the investigation. The
purpose of a SOP is to carry out the operations
correctly and always in the same manner. A SOP
should be available at the place where the work is
done".
A SOP is a compulsory instruction. If deviations
from this instruction are allowed, the conditions
for these should be documented.
 Fundamental SOPs: These give instructions
how to make SOPs of the other categories.
 Methodic SOPs: These describe a complete
testing system or method of investigation.
 SOPs for safety precautions.
 SOPs for operating instruments, apparatus
and other equipment.
 SOPs for analytical methods.
 SOPs for the preparation of reagents.
 SOPs for receiving and registration of
samples.
 SOPs for Quality Assurance.
 SOPs for archiving and how to deal with
complaints.
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Good Manufacturing
Practices
Good manufacturing practices are the sets of the
principles, regulations, codes (law or official
standard), guidelines and procedures and part of
quality assurance system which must be followed
by the manufacturers to ensure that the products that
are consistently produce are of quality standard and
appropriate for their intended use and cover the
manufacturing and testing of pharmaceutical dosage
form and active pharmaceutical ingredients,
diagnostics, foods, various other pharmaceutical
products and medical devices
GMP Principles:
Good manufacturing practice guidelines provides
guidance for manufacturing, testing, and quality
assurance in order to ensure that drug product is
safe for human consumption. Many countries have
legislatsed that pharmaceutical and medical device
manufacturer must follow GMP procedures, and
have created their own GMP guidelines that
correspond with their legislation.
All guidelines follow a few basic principles:
 Hygiene: Pharmaceutical manufacturing facility
must maintain a clean and hygienic
manufacturing area.
 Controlled environmental conditions in order to
prevent cross contamination of drug product
from other drug or extraneous particulate matter
which may render the drug product unsafe for
human consumption
 Manufacturing processes are clearly defined and
controlled. All critical processes are validated to
ensure consistency and compliance with
specifications.
 Manufacturing processes are controlled, and any
changes to the process are evaluated. Changes
that have an impact on the quality of the drug
are validated as necessary.
 Instructions and procedures are written in clear
and unambiguous language.
Adnan Sarwar Chaudhary
 Operators are trained to carry out and document
procedures.
 Records are made, manually or by instruments,
during manufacture that demonstrate that all the
steps required by the defined procedures and
instructions were in fact taken and that the
quantity and quality of the drug was as
expected. Deviations are investigated and
documented.
 Records of manufacture that enable the
complete history of a batch to be traced are
retained in a comprehensible and accessible
form.
 The distribution of the drugs minimizes any risk
to their quality.
 A system is available for recalling any batch of
drug from sale or supply.
 Complaints about marketed drugs are examined,
the causes of quality defects are investigated,
and appropriate measures are taken with respect
to the defective drugs and to prevent recurrence.
Current Good
Manufacturing Practices
The cGMP regulations establish requirements for all
aspects of pharmaceutical manufacture. They apply
to domestic and to foreign suppliers and
manufacturers whose bulk components and finished
pharmaceutical products are imported, distributed,
or sold in this country. To ensure compliance, the
FDA inspects the facilities and production records
of all firms covered by these regulations.
The Code of Federal Regulations (CFR) contains
(a) requirements for the ―Current Good
Manufacturing Practice for Finished
Pharmaceuticals‖ and (b) additional cGMP
requirements for biologic products, (c) medicated
articles, and (d) medical devices. Currency and
compliance with cGMP regulations are supported
through notices in the Federal Register and through
the FDA‘s Compliance Policy Guide and various
other guidance‘s issued by the FDA.
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Current Good
Manufacturing Practice for
Finished Pharmaceuticals
General Provisions: Scope and definitions
The regulations in 21 CFR, part 211, contain the
minimum GMP requirements for the preparation of
finished pharmaceutical products for administration
to humans or animals. Common terms used in these
regulations are defined as follows:
Active ingredient or active pharmaceutical
ingredient (API): Any component that is intended to
furnish pharmacologic activity or other direct effect
in the diagnosis, cure, mitigation, treatment, or
prevention of disease or to affect the
structure or function of the body of man or other
animals
Batch: A specific quantity of a drug of uniform
specified quality produced according to a single
manufacturing order during the same cycle of
manufacture
Batch wise control: The use of validated in process
sampling and testing methods in such a way that
results prove that the process has done what it
purports to do for the specific batch concerned
Certification: Documented testimony by qualified
authorities that a system qualification, calibration,
validation, or revalidation has been performed
appropriately and that the results are acceptable
Compliance: Determination through inspection of
the extent to which a manufacturer is acting in
accordance with prescribed regulations, standards,
and practices
Component: Any ingredient used in the
manufacture of a drug product, including those that
may not be present in the finished product
Drug product: A finished form that contains an
active drug and inactive ingredients. The term may
also include a form that does not contain an active
ingredient, such as a placebo.
Inactive ingredient: Any component other than the
active ingredients in a drug product
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Lot: A batch or any portion of a batch having
uniform specified quality and a distinctive
identifying lot number
Lot number, control number, or batch number: Any
distinctive combination of letters, numbers, or
symbols from which the complete history of the
manufacture, processing, packaging, holding, and
distribution of a batch or lot of a drug
product may be determined
Master record: Record containing the formulation,
specifications, manufacturing procedures, quality
assurance requirements, and labeling of a finished
product
Quality assurance: Provision to all concerned the
evidence needed to establish confidence that the
activities relating to quality are being performed
adequately
Quality audit: A documented activity performed in
accordance with established procedures on a
planned and periodic
basis to verify compliance with the procedures to
ensure quality
Quality control: The regulatory process through
which industry measures actual quality
performance, compares it with standards, and acts
on the difference
Quality control unit: An organizational element
designated by a firm to be responsible for the duties
relating to quality control
Quarantine: An area that is marked, designated, or
set aside for the holding of incoming components
prior to acceptance testing and qualification for use
Representative sample: A sample that accurately
portrays the whole
Reprocessing: The activity whereby the finished
product or any of its components is recycled
through all or part of the manufacturing process
Strength: The concentration of the drug substance
per unit dose or volume
Verified: Signed by a second individual or recorded
by automated equipment
Validation: Documented evidence that a system
(e.g., equipment, software, controls) does what it
purports to do
Process validation: Documented evidence that a
process (e.g., sterilization) does what it purports to
do
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Validation protocol: A prospective experimental
plan to produce documented evidence that the
system has been validated
Organization and Personnel
The organization and personnel section of the
regulations deals with the responsibilities of the
quality control unit, employees, and consultants.
The regulations require that a quality control unit
have the authority and responsibility for all
functions that may affect product quality. This
includes accepting or rejecting product components,
product specifications, finished products,
packaging, and labeling. Adequate laboratory
facilities shall be provided, written procedures
followed, and all records maintained.
All personnel engaged in the manufacture,
processing, packing, or holding of a drug product,
including those in supervisory positions, are
required to have the education, training, and/or
experience needed to fulfill the assigned
responsibility. Appropriate programs of skill
development, continuing education and training,
and performance evaluations are essential for
maintaining quality assurance. Any consultants
advising on scientific and technical matters should
possess requisite qualifications for the tasks.
Buildings and facilities
The regulations in this section include the design,
structural features, and functional aspects of
buildings and facilities. Each building‘s structure,
space, design, and placement of equipment must be
such to enable thorough cleaning, inspection, and
safe and effective use for the designated operations.
Proper considerations must be given to such factors
as water quality standards; security; materials used
for floors, walls, and ceilings; lighting; segregated
quarantine areas for raw materials and product
components subject to quality control approval;
holding areas for rejected components; storage areas
for released components; weighing and measuring
rooms; sterile areas for ophthalmic and parenteral
products; flammable materials storage areas;
finished products storage; control of heat, humidity,
temperature, and ventilation; waste handling;
employee facilities and safety procedures in
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compliance with the Occupational Safety and
Health Administration regulations; and procedures
and practices of personal sanitation.
All work in the manufacture, processing, packaging,
or holding of a pharmaceutical product must be
logged in, inspected by a supervisor, and signed off.
Similarly, a log of building maintenance must be
kept to document this component of the regulations.
Equipment
Each piece of equipment must be of appropriate
design and size and suitably located to facilitate
operations for its intended use, cleaning, and
maintenance. The equipment‘s surfaces and parts
must not interact with the processes or product‘s
components so as to alter the purity, strength, or
quality.
Standard operating procedures must be written and
followed for the proper use, maintenance, and
cleaning of each piece of equipment, and
appropriate logs and records must be kept.
Automated equipment and computers used in the
processes must be routinely calibrated, maintained,
and validated for accuracy.
Filters used in the manufacture or processing of
injectable drug products shall not release fibers into
such products. If fiber-releasing filters must be
used, non–fiber-releasing filters also must be used
to reduce any fiber content.
Control of Components, Containers, and Closures
Written procedures describing the receipt,
identification, storage, handling, sampling, testing,
and approval or rejection of all drug product
components, product containers, and closures must
be maintained and followed. Bulk pharmaceutical
chemicals, containers, and closures must meet the
exact physical and chemical specifications
established with the supplier at the time of ordering.
When product components are received from a
supplier, each lot must be logged in with the
purchase order number, date of receipt, bill of
lading, name and vital information of the supplier,
supplier‘s stock or control number, and quantity
received. The component is assigned a control
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number that identifies both the component and the
intended product. Raw materials are quarantined
until they are verified through representative
sampling and careful qualitative and quantitative
analysis. The quality control unit approves and
releases for use in manufacture only those that meet
the specifications. The assigned control number
follows the component throughout production so it
can be traced if necessary.
Rejected components, drug product containers, and
closures are identified and controlled under a
quarantine system to prevent their use in
manufacturing and processing operations. As the
majority of bulk chemicals (APIs) are synthesized
overseas (primarily in China and India), it is
important to confirm their identity and purity and
conformance with United States Pharmacopeia
(USP) and National Formulary (NF) standards prior
to use in finished pharmaceuticals.
Production and Process Controls
Written procedures are required for production and
process controls to ensure that the drug products
have the correct identity, strength, quality, and
purity. These procedures, which include the charge-
in of all components, use of in-process controls,
sample testing, and process and equipment
validation, must be followed for quality assurance.
Any deviation from the written procedures must be
recorded and justified. In most instances, the
operator records time and date of each key
operation, and the supervisor signs off on it. When
operations are controlled by automated equipment,
such equipment must be validated regularly for
precision.
All product ingredients, equipment, and drums or
other containers of bulk finished product must be
distinctively identified by labeling as to content
and/or status. In process samples are taken from
production batches periodically for product control.
In process controls are of two general types: (a)
those performed by production personnel at the time
of operation to ensure that the machinery is
producing output within preestablished control
limits (e.g., tablet size, hardness) and (b) those
performed by the quality control laboratory
Adnan Sarwar Chaudhary
personnel to ensure compliance with all product
specifications (e.g., tablet content, dissolution) and
batch-to-batch consistency. Product found out of
standard sometimes may be reprocessed for
subsequent use. However, in this, as in all instances,
procedures must be performed according to
established protocol, and all materials must be
accounted for, all specifications met, and all records
meticulously maintained.
Packaging and labeling Control
Written procedures are required for the receipt,
identification, storage, handling, sampling, and
testing of drug product and issuance of labeling and
packaging materials. Labeling for each variation in
drug product—strength, dosage form, or quantity of
contents—must be stored separately with suitable
identification. Obsolete and outdated labels and
other packaging materials must be destroyed.
Access to the storage area must be limited to
authorized personnel.
All materials must be withheld for use in the
packaging and labeling of product until approved
and released by the quality control unit. Control
procedures must be followed and records
maintained for the issuance and use of product
labeling. Quantities issued, used, and returned must
be reconciled and discrepancies investigated. Before
labeling operations commence, the labeling
facilities must be inspected to ensure that all drug
products and labels have been removed from the
previous operations. There must be dedication of
labeling and packaging lines to each different
strength of each different drug product. There must
be use of appropriate electronic or
electromechanical equipment to conduct a 100%
examination for correct labeling during or after
completion of finishing operations or use of visual
inspection to conduct a 100% examination for
correct labeling during or after completion of
finishing operations for hand-applied labeling. Such
examination shall be performed by one person and
independently verified by a second person. All of
these procedures are essential to avoid label mix-
ups and the mislabeling of products. All records of
inspections and controls must be documented in the
batch production records.
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Expiration Dating
To assure that a drug product meets applicable
standards of identity, strength, quality, and purity at
the time of use, it must bear an expiration date
determined by appropriate stability testing. Exempt
from this requirement are homeopathic drug
products, allergenic extracts, and investigational
drugs that meet the standards established during
preclinical and clinical studies.
Tamper-Evident Packaging
On November 5, 1982, the FDA published initial
regulations on tamper-resistant packaging in the
Federal Register. These regulations were
promulgated after criminal tampering
with over-the-counter (OTC) drug products earlier
in that year resulted in illness and deaths. In the
primary incident, cyanide was surreptitiously placed
in acetaminophen capsules in commercial packages.
Today, the cGMP regulations require tamper-
evident packaging for OTC drug products to
improve their security and to assure their safety and
effectiveness. All OTC drug products offered for
retail sale are required to have tamper-evident
packaging except for some categories, such as
dentifrices, dermatologicals, insulin, and throat
lozenges. For other product categories, a
manufacturer may file with the FDA a Request for
Exemption from Tamper-Evident Rule. The petition
is required to contain specific information on the
drug product, the reasons the requirement is
unnecessary or cannot be achieved, and alternative
steps the petitioner has taken or may take to reduce
the likelihood of malicious adulteration of the
product. Generally exempt from these regulations
are products not packaged for retail sale but rather
distributed to hospitals, nursing homes, and health
care clinics for institutional use.
A tamper-evident package is defined as ―one having
one or more indicators or barriers to entry which, if
breached or missing, can reasonably be expected to
provide visible evidence to consumers that
tampering has occurred‖. The indicators or barriers
may involve the immediate drug product container
and/or an outer container or carton. For two-piece
hard gelatin capsule products, a minimum of two
Adnan Sarwar Chaudhary
tamper-evident packaging features is required
unless the capsules are sealed with tamper-resistant
technology.
Even with these safeguards in effect, the possibility
of drug product tampering requires the pharmacist
and consumer to remain constantly vigilant for
signs of product entry. Pharmaceutical
manufacturers have the option of determining the
type of tamper resistant packaging to use.
Holding and distribution
Written procedures must be established and
followed for the holding and distribution of product.
Finished pharmaceuticals must be quarantined in
storage until released by the quality control unit.
Products must be stored and shipped under
conditions that do not affect product quality.
Ordinarily, the oldest approved stock is distributed
first. The distribution control system must allow the
distribution point of each lot of drug product to be
readily determined to facilitate its recall if
necessary.
Laboratory Controls
Laboratory controls are requirements for the
establishment of and conformance to written
specifications, standards, sampling plans, test
procedures, and other such mechanisms. The
specifications, which apply to each batch of drug
product, include provisions for sample size, test
intervals, sample storage, stability testing, and
special testing requirements for certain dosage
forms, including parenterals, ophthalmics,
controlled-release products, and radioactive
pharmaceuticals.
Reserve samples must be retained for distributed
products for specified periods depending on their
category. Animals used in testing components, in-
process materials, or drug products for compliance
with established specifications shall be maintained
and controlled in a manner that assures their
suitability for their intended use. They shall be
identified, and adequate records shall be maintained
showing the history of their use. records and reports
Production, control, and distribution records
must be maintained for at least a year following the
expiration date of a product batch. This includes
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equipment cleaning and maintenance logs;
specifications and lot numbers of product
components, including raw materials and product
containers and closures; and label records.
Complete master production and control records for
each batch must be kept.
These master records must document that
each step in the production, control, packaging,
labeling, and distribution of the product was
accomplished and approved by the quality control
unit. Depending on the operation, the operator‘s
and/or supervisor‘s full signatures, initials, or other
written or electronic identification codes are
required.
Records of written and oral complaints regarding a
drug product (e.g., product failure, adverse drug
experience) must also be maintained, along with
information regarding the internal disposition of
each complaint. All records must be made available
at the time of inspection by FDA officials.
Returned and Salvaged drug Products
Returned drug products (e.g., from wholesalers)
must be identified by lot number and product
quality determined through appropriate testing.
Drug products that meet
specifications may be salvaged or reprocessed.
Those that do not, along with those that have been
subjected to improper storage (e.g., extremes in
temperature), shall not be returned to the
marketplace. Records for all returned products must
be maintained and must include the date and
reasons for the return; quantity and lot number of
product returned; procedures employed for holding,
testing, and reprocessing the product; and the
product‘s disposition.
Information technology and automation
Although not part of the cGMP requirements,
the effective deployment of information
technologies and automated systems can enhance
pharmaceutical process development,
production efficiencies, product quality, and
regulatory compliance.
Adnan Sarwar Chaudhary
Computers are used extensively in plant
operations such as production scheduling, in-
process manufacturing, quality control, and
packaging and labeling. The networking of
computers in the production and quality control
areas fully integrates laboratory information and
manufacturing operations into sophisticated
management systems. These integrated systems
support cGMP compliance, process validation,
resource management, and cost control. Robotic
devices increasingly are being employed to replace
manual operations in production lines, analytical
sampling, and packaging.
Laboratory robotics provides automation in
areas such as sample preparation and handling, wet
chemistry procedures, laboratory process control,
and instrumental analysis. Pharmaceutical
applications of robotics include automated product
handling in production lines and in procedures such
as sampling and analysis, tablet content uniformity,
and dissolution testing.
Validation
The FDA defines validation as:
Establishing documented evidence which provides a
high degree of assurance that a specific process or
device will consistently produce data or a product
that meets its predetermined specifications and
quality attributes.
OR
The documented act of demonstrating that any
procedure, process and activity will consistently
lead to the expected results.
OR
Validation of any process is a scientific
demonstration by appropriate tests that the process
actually accomplishes the intended effect under
specified operating conditions.
It includes the qualification of systems and
equipments. For example validation of Sterilization
process.
Types of Validation
i. Prospective Validation
ii. Retrospective Validation
iii. Concurrent Validation
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i. Prospective Validation
Prospective validation is conducted before the Accuracy Precision
distribution of either a new product or a product Definition: ―The ability of a
made under a modified production process. It is a ―The ability of a measurement to be
preplanned scientific approach and includes: measurement to match consistently
the actual value of the reproduced.‖
 The Initial Stages Of Formulation
quantity being OR
Development
measured.‖ ―The number of
 Process Development OR significant digits to
 Setting Of A Process Specifications The accuracy of an which
 Sampling Plans analytical method is a value has been reliably
 Designing Of Batch Records the extent to which test measured.‖
 Defining Raw Material Specifications results generated by
 Completion Of Pilot Runs the method and the
 Transfer Of Technology From Scale Up true value agree.
Batches To Commercial Size Batches
 Listing Major Process Equipment Example: If on several tests the
If the temperature temperature sensor
 Environmental control
outside is 34.0 F and a matches the actual
ii. Retrospective Validation temperature sensor temperature while the
Retrospective validation is conducted for a product also reads 34.0 F, actual temperature held
already being marketed and is based on extensive then the sensor is constant, then the
data accumulated over several slots. Retrospective accurate. sensor is precise.
validation may be used by the older products which By second definition:
were not validated at the time that they were first the no. 4.1215 is
marketed and which is now to be validated to more precise than the
conform to the requirements of FDA. no. 4.12.

iii. Concurrent Validation Accuracy is how close Precision is how

It is the process in which current production batches to true/actual consistent your results
are used to monitor processing parameters. It gives measurement are. are
assurance of the present batch being studied and for the same phenomena
offers limited assurance regarding consistency of over several
quality from batch to batch. measurements.

Accuracy Single factor or Multiple measurements

The accuracy of a measurement system is the measurement or factors are
degree of closeness of measurements of a quantity needed
to its actual (true) value.

Precision
The precision of a measurement system, also called
reproducibility or repeatability, is the degree to
which repeated measurements under unchanged
conditions show the same results.
Sensitivity
Capacity of test procedure or an instrument to
record small variations in concentrations is called
sensitivity.
Selectivity
Ability of method to measure accurately and
specifically the analyte of interest even in the

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presence of matrix and other components like
impurities in the sample is called selectivity.
Linearity
Ability of method to produce test results that are
directly proportional to concentration of analyte is
called linearity. The linearity of a method gives the
characteristic trend of parameters such as
absorbance, peak height, and peak area or response
ratio as a function of concentration of the
component to be measured.
Range
Lowest and highest level of analyte that a method
can determine with reasonable accuracy and
precision is called its range.
The range is normally expressed in the same units
as the test results e.g. percentage, parts per million
obtained by the analytical method.
Limit of Detection (LOD)
Lowest concentration of analyte in a sample that a
method can detect but doesn‘t necessarily quantify
understated experimental conditions. It simply
indicates that analyte in the sample is below or
above certain level.
Limit of Quantitation (LOQ)
LOQ is the lowest concentration of a substance in a
sample that can be estimated quantitatively with
acceptable precision and accuracy.
Ruggedness
It is the degree of reproducibility of obtained by
analyzing same sample under variety of normal test
conditions, different analysts, instruments, reagents
and days.
Robustness
It is the measurement of capacity of a method to
remain unaffected by small but considerable
variations in procedure.
Process Re-Validation
It is required when there is a change in any of the
critical process parameters, formulation, primary
packaging components, raw material fabricators or
major equipment. Failure to meet product or process
Adnan Sarwar Chaudhary
specifications in sequential batches would also
require process re-validation.
Qualification
Qualification is a process of assurance that the
specific system, premises or equipment are able to
achieve the predetermined acceptance criteria to
confirm the attributes what it aims to do.
Calibration
The comparison of the measurement system or
device of unknown accuracy to another
measurement system or device with a known
accuracy to detect, correlate, report or eliminate any
variation is called calibration. Calibration is done to
ensure that measuring equipment used in a
manufacturing process or analytical procedure gives
measurements that are correct within established
limits. For example calibration of a pH meter.
 Things are qualified. (For example equipments,
systems etc).
 Processes and procedures are validated. (For
example sterilization process).
 Calibration is one of the condition/step for
qualification whereas Qualification is one of the
condition/step for validation.
Clean areas & rooms
Area with defined environment control of particulate and
microbial contamination constructed and used in such
a way as to reduce the introduction, generation and
retention of contamination within the area.
It is divided into 3 classes:
Class 100 area
Area not exceded from 100 particles/cubic feet of
0.5µ size.
Class 1000 area
Area not exceded from 1000 particles/cubic feet of
0.5µ size
Class 100,000 area
Area not exceded from 100,000 particles/cubic feet
of 0.5µ size.
Test
―A technical operation that consists of
determination of one or more characteristics or
performance of a given product, material,
equipment, organism, process or service according
to a specified procedure.‖
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In Process Quality Control (IPQC)
In process quality control is a process of monitoring
critical variables of manufacturing process to ensure
a quality of the final product. In-process
manufacturing controls are established and
documented by quality control and production
personnel to ensure that quality of the product is
within the acceptable standard range.
Statistical Quality Control (SQC)
―The monitoring of quality by application of
statistical methods in all stages of production.‖
Or
―The application of statistical methods to quality
control.‖
It refers to characteristics of product from both
quantitative and qualitative point of view to meet
established standards.
By using charts and collecting few frequent
samples, we can detect the change in process that
may affect the quality.
These charts are:-
 Control charts for variables
 Control charts for attributes
Dissolution Test:
―Dissolution testing is required for all solid oral
Pharmacopoeial dosage forms in which absorption
of the drug is necessary for the product to exert the
desired therapeutic effect. Exceptions are for tablets
meeting a requirement for completeness of solution
or for rapid (10 to 15 minutes) disintegration for
soluble or radiolabeled drugs.‖
Laminar Flow Hood:
Laminar flow is unidirectional air moving at a
steady velocity along parallel lines. Laminar flow
cabinets may or may not be biological safety
cabinets.
OR
The laminar flow hood provides an aseptic work
area while allowing the contaminant of infectious
splashes or by many microbiological procedures.
HEPA Filter:
It is designed to remove particles, including
microorganisms, from the air. HEPA filters are
effective at trapping particulates & infectious agents
Adnan Sarwar Chaudhary
but not at capturing volatile chemicals or gas. Only
certain classes of biological safety cabinets that are
exhausted to the outside can be used when working
with small amounts of volatile chemicals.
Pyrogen Testing
1. Rabbit Test:
This test involves measurement of rise in body
temperature of rabbits following the IV injection of
a sterile solution of a substance to be examined. It is
designed for products that can be tolerated by test
rabbit in a dose not exceeding 10ml per kg injected
with in a period of not more than 10 minutes.
2. Lal’s Test:
Limulus amebocyte lysate (LAL) is an aqueous
extract of blood cells (amoebocytes) from the horse
shoe crab, Limulus polyphemus. LAL reacts with
bacterial endotoxin or lipopolysaccharide (LPS),
which is a membrane component of Gram negative
bacteria. This reaction is the basis of the LAL test,
which is used for the detection and quantification of
bacterial endotoxins.
Sterility:
Sterility or freedom from the presence of viable
microorganisms, in a strict, uncompromising
requirement of an injectable dosage form.
Rheology:
Rheology is the study of the flow of matter,
primarily in the liquid state, but also as 'soft solids'
or solids under conditions in which they respond
with plastic flow rather than deforming elastically
in response to an applied force.
It applies to substances which have a complex
microstructuring like muds, sludges, suspensions,
polymers and other glass formers (e.g., silicates), as
well as many foods and additives, bodily fluids
(e.g., blood) and othe biological materials or other
materials which belong to the class of soft matter.
Pyrogens:
Pyrogen consists of two words:
 Pyro-pyrexia (fever or rise in body temp.)
 Gen-producing or generation.
―Pyrogen is simply fever producing agent.‖
―Pryrogens are substances that cause febrile
reactions when sufficient amount enters in
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circulatory system.‖ Bacterial endotoxin is the most
significant pyrogen because of its potency and
ubiquity.
Friability:
―Friability (the condition of being friable) is the
ability of a solid substance.‖
A friable substance is any substance that can be
reduced to fibers or finer particles by the action of a
small pressure or friction.
management, improving the customer experience
and ensuring that employees are up-to-speed with
their training. Total quality management aims to
hold all parties involved in the production process
as accountable for the overall quality of the final
product or service.‘‘
Weight Variation
This test is performed to check that the tablet
contains the proper amount of drug and to ensure
that the dose is in safe therapeutic window.

Stability Studies: Uniformity Of Content

Stability studies are a critical part of the drug Uniformity of Content is a pharmaceutical analysis
development process and are essential for drug technique for the quality control of capsules or
product marketing approval. Stability studies are tablets. Multiple capsules or tablets are selected at
conducted at all phases of the drug development random and a suitable analytical method is applied
cycle for different purposes with the ultimate goal to assay the individual content of the active
of having a stable product on the market. During ingredient in each capsule or tablet. It is a test to
development, stability studies are conducted to ensure homogenous distribution of active ingredient
support the formulation development and safety and in a dosage form. It involves weighting, crushing,
efficacy claims of investigational new drugs. making dilutions and analyzing by using a
Difference Between Iodometry And Iodimetry: recommended analysis method.
Iodometry Iodimetry
1. When an analyte 1. When an analyte that
that is an oxidizing is a reducing agent is
agent titrated directly with a
is added to excess standard iodine solution,
iodide to produce the method is called
iodine, and the iodine "iodimetry"
produced is 2. Iodimetry, a species is
determined by titration directly titrated with an
with sodium iodine solution.
thiosulfate, the method 3. Iodimetry is a direct
is called method.
"iodometry". 4. Iodimetry can be used
2. Iodometry a species to quantify reducing
is titrated with an agents.
iodide solution and
then the released
iodine is titrated with
thiosulphate.
3. Iodometry is an
indirect method.
4. Iodometry can be
used to quantify
oxidizing agents

Total Quality Management (TQM)

―The continuous process of reducing or eliminating
errors in manufacturing, streamlining supply chain
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QALITY
CONTROL OF
SOLID DOSAGE
FORMS

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Dosage Forms:
Completed forms of the pharmaceutical preparation
in which prescribed doses of medication are
included.
They are designed to:
 Resist action by gastric fluids.
 Prevent Vomiting and Nausea
 Reduce or alleviate the undesirable taste and
smell associated with oral administration.
 Achieve a high concentration of drug at
target site.
 Produce a delayed or long-acting drug effect.
Solid Dosage Forms:
Solid dosage forms include
 Tablets
 Capsules
 Granules
 Powders
Tablets:
Tablets are solid dosage forms containing one or
more active ingredients. They are unit dosage form.
They are obtained by single or multiple
compression and may be coated or uncoated. They
are usually intended for oral applications but
sometime they also have some alternative
applications such as implants, tablets for injection,
irrigation or external use, vaginal tablets etc.
Capsules:
Capsules are solid dosage forms with hard or soft
gelatin shells. They are of various shapes and sizes;
contain a single dose of one or more ingredients.
They are intended for oral administration.
Powders:
A powder is intimate mixture of dry, finely divided
drugs and/or chemicals that may be intended for
internalor external use.
Granules:
Granule is generic term used for small particle or
grain, A granule is formed when small particles
Adnan Sarwar Chaudhary
gathered into longer, permanent aggregate in which
the original particles can still be identified.
GENERAL APPEARANCE
Different parameters of tablet appearance are
 Size and Shape
 Color
 Odor
 Surface Texture
1. Size and Shape:
Purpose:
To ensure the trouble free packing improve
elegance and to control the weight of tablet.
Instruments:
 The size of tablet is directly related to
thickness and diameter of tablets, that depends
on the size of die and punches, amount of fill
material and force /pressure of compression.
 The thickness and diameter of the tablet may
be measured manually (by micrometer screw
gauge and venire calipers) or by automatic
equipment.
Procedure:
10 tablets are taken randomly to measure the
thickness and diameter by placing them vertically
and horizontally in the Jaws of instruments.
For uncoated tablets
 The diameter should be 4mm-14mm
 The thickness should be 2mm-4mm
 Tablets having the diameter less than
12.5mm have variation of ±5%
 Tablets having the diameter more than
12.5mm have variation of ±3%
Specification:
±5% of stated to tablet hardness and can be used as
initial control during production.
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Advantages: Official Tests:

 Thickness of tablets is directly related to tablet
1. Disintegration
hardness and can be used as initial control
2. Weight Variation
parameter.
3. Friability
 Limits of thickness and diameter of tablet
4. Hardness
depend on tablet weight to ensure trouble free
packaging (when tablets are thicker than Non-Official Tests:
normal then packaging process is disturbed).
 The shape of tablet should be as provided by 1. Thickness & Diameter
monograph i.e. round, circular, double convex
etc and for shaped tablets slotted punches must Hardness (BP) Or Breaking
be run at slower speeds.

Application:
Force (USP)
 Ensure the trouble free packaging. This test is intended to determine, under defined
 Used as control parameter. conditions, the resistance to crushing of tablets,
measured by the force needed to disrupt them by
2. Color: crushing.
 Color specification is helpful for the It is the resistance of tablet against applied force till
identification of specific product during it breaks.
manufacturing process. OR
 There should be equal distribution of color as It is the load required to crush the tablet when
uniform distribution of color increases the placed on its edge.
ethical appeal.
 Sometime during manufacturing the problem
of mottling (unequal and uneven distribution of Importance
color) occur that shows poor quality product. Check the breakage during

 Manufacture
3. Odour:
 Packaging
 Presence or absence of odor in a batch of tablet
 Handling
is also tested.
 Storage
 Some drugs produce characteristic odor e.g.
multivitamins and it is helpful for the detection Why do we measure hardness?
of material.  To determine the need for pressure
 While in some drugs presence of odour show adjustments on the tableting machine.
the unstable drug or degradation e.g. aspirin has  To determine the disintegration time.
vinegar like odour if degraded.  To determine elasticity.

4. Surface texture: Factors Affecting the Hardness:

 The surface of tablet should be smooth and
there must be no chips, cracks, contamination
from external substances, capping and
sticking.
Physical Tests
 Compression of the tablet and compressive
force.
 Amount of binder. (More binder a more
hardness)
 Method of granulation in preparing the
tablet (wet method gives more hardness than

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direct method; Slugging method gives the
best hardness).
Types of hardness testers:
i. Manual or mechanical
1. Strong-Cobb tester
2. Monsanto Hardness tester
3. Eureka Hardness tester
4. Pfizer Hardness tester
ii. Motor driven
1. Heberlein Schleuniger Hardness tester
2. Erweka Hardness tester
3. Casburt Hardness tester
Procedure:
i. Stoker-Monsanto Hardness Tester (manual):
 It is a small potable hardness tester,
manufactured by Monsanto chemicals.
 Select 6 tablets randomly according to USP
 Place the tablets, diametrically, between the
moving and fixed jaw one by one.
 Adjust the reading of indicator scale to zero.
 Gradually increase the force applied to the
edge of tablet by moving the screw knob
forward until the tablet breaks.
 The reading is noted from the scale which
indicates the pressure required in kg to break
the tablet.
ii. Eureka Hardness Tester (mechanical):
 In this instrument the breaking force is
applied by a beam fastened to one end to a
pivot.
 The motor moves a weight along the beam
at a constant speed and increase the force
against the tablet.
 When the tablet breaks, a micro switch is
activated that stop the motor.
 An indicator is fastened to the weight shows
the breaking strength on a scale in kg.
 The average hardness acceptable for
compressed tablet ranges between 5-10
kg/cm2.
 German mode instrument has scale in
Newton and 1N=9.8kg.
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Acceptance criteria
5-10 kg/cm2 according to USP
i. According to USP:
Minimum of 6 tablets should be tested
ii. According to BP:
Carryout the measurement on each tested tablet.
Thickness Diameter
Devices:
Micrometer Screw gauge, Vernier calipers or
automatic equipments
Significance:
Proper packaging of solid dosage form, i.e. in
blister, strip, bulk or bottle packagings.
Factors Affecting the thickness & diameter:
Following factors are there:
 Tablet compression or force
 Amount of material in punch or die
 Depth and diameter of die Procedure
 Select 10 tablets from the batch randomly
and measure the thickness and diameter by
the devices.
 All the tablets must follow the acceptance
criteria.
Allowed variation:
 Allowed range = ±5%
Acceptance criteria
 Thickness = 2-4mm
 Diameter = 4-14mm
Friability
It is the tendency of tablets to powder, chip, or
fragment and this can affect the elegance
appearance, consumer acceptance of the tablet, and
also add to tablet’s weight variation or content
uniformity problems.
OR
It is the resistance of the tablets against the
mechanical shock during packaging, handling and
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transportation and helpful for checking the
lamination and capping.
Significance
 Check breakability
 Check drug loss
 Check capping and hardness
Apparatus:
An instrument called friabilator consisting of drum
is used to evaluate the ability of the tablet to
withstand abrasion in packaging, handling, and
shipping.
 Internal diameter 283-291mm
 Depth 36-40mm
 Made of transparent synthetic polymer with
internal surface polished.
 A curved projection with an inside radius
b/w 75.5-85.5mm that extent from middle of
the drum to outer wall from where tablets
are tumbled.
 Drum is rotated at 25± 1rpm.
 Thus at turn, tablets rolls or slide, and fall
onto the drum wall or onto each other.
Uncoated Tablets
This chapter provides guidelines for the friability
determination of compressed, uncoated tablets. The
test procedure presented in this chapter is generally
applicable to most compressed tablets.
Measurement of tablet friability supplements other
physical strength measurements, such as tablet
breaking force.
Use a drum, with an internal diameter between 283-
291 mm and a depth between 36-40 mm, of
transparent synthetic polymer with polished internal
surfaces, and subject to minimum static build-up
(see Figure.). One side of the drum is removable.
The tablets are tumbled at each turn of the drum by
a curved projection with an inside radius between
75.5-85.5 mm that
extends from the middle of the drum to the outer
wall. The outer diameter of the central ring is
between 24.5-25.5 mm. The drum is attached to the
Adnan Sarwar Chaudhary
horizontal axis of a device that rotates at 25 ± 1
r/min. Thus, at each turn the tablets roll or slide and
fall onto the drum wall or onto each other.
For tablets with a unit mass equal to or less than
650 mg, take a sample of whole tablets
corresponding as near as possible to 6.5 g. For
tablets with a unit mass of more than 650 mg, take
a sample of 10 whole tablets. The tablets are
carefully dedusted prior to testing. Accurately
weigh the tablet sample, and place the tablets in the
drum. Rotate the drum 100 times, and remove the
tablets. Remove any loose dust from the tablets as
before, and accurately weigh.
Generally, the test is run once. If obviously cracked,
cleaved, or broken tablets are present in the tablet
sample after tumbling, the sample fails the test. If
the results are difficult to interpret or if the weight
loss is greater than the targeted value, the test is
repeated twice and the mean of the 3 tests
determined. A maximum loss of mass (obtained
from a single test or from the mean of 3 tests) not
greater than 1.0 per cent is considered acceptable
for most products
If tablet size or shape causes irregular tumbling,
adjust the drum base so that the base forms an angle
of about 10° with the horizontal and the tablets no
longer bind together when lying next to each other,
which prevents them from falling freely.
Effervescent tablets and chewable tablets may have
different specifications as far as friability is
concerned. In the case of hygroscopic tablets, a
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humidity-controlled environment is required for compressed air to 0.45 m3·h-1. After 15 min, remove
testing. the granules or spheroids from the apparatus by
A drum with dual scooping projections, or disconnecting the U-tube and weigh again (m2).
apparatus with more than one drum, for the running Test 3 samples and calculate the mean value. It is
of multiple samples at one time, are also permitted. recommended to spray the inside of the apparatus
with an antistatic agent every 3 determinations in
Granules and Spheroids order to prevent electrostatic charging.
This chapter describes 2 methods for determination
Loss on drying Dry in an oven at 105 °C, unless
of the friability of granules and spheroids, which
otherwise prescribed. Alternatively, other
may be used during development studies. It is
drying conditions as described in general method
recognised, however, that many methods with equal
may be used.
suitability may be used.
Calculation
This test is intended to determine, under defined
conditions, the friability of granules and
spheroids. Friability is defined as a reduction in the
mass of the granules or spheroids or in the
formation of fragments of granules or spheroids, F = Friability;
occurring when the granules or spheroids are T1 = Percentage loss on drying before the test (mean
subjected to mechanical strain during handling of 2 determinations);
(tumbling, vibration, fluidisation, etc.). Examples T2 = Percentage loss on drying after the test (mean
of changes are abrasion, breakage or deformation of of 2 determinations);
granules or spheroids. m1 = Mass of the granules or spheroids before the
test, in grams;
Method A Apparatus (fluidised-bed apparatus) m2 = Mass of the granules or spheroids after the test,
The apparatus (see Figure) consists of a glass in grams.
cylinder (A) with a conical lower part. The cylinder
is provided with a sieve lid (B) having an aperture
size of 500 µm or any other suitable sieve. The
conical end is connected to a U-shaped glass tube
(C) that can be disconnected from the cylinder for
removal of the granules or spheroids. The U-tube is
attached to a T-coupling (D). One inlet of the T-
coupling is joined by a silicone tube to a manometer
for regulating the compressed-air flow (use
compressed air complying with the test for water in
the monograph Medicinal air (1238)), the other one
is
connected via a silicone tube to a by-pass flow
meter (E) (0.10-1.00 m3·h-1).

Procedure
The following procedure is usually suitable.
Remove the fine particles by sieving(sieve having
an aperture size of 710 µm or any other suitable
sieve). Introduce about 8.0 g (m1) of granules or
spheroids into the cylinder (A). Close the apparatus
with the sieve lid (B). Adjust the flow rate of the
Method B
Apparatus (oscillating apparatus)
The apparatus (see Figure 2.9.41.-2) consists of a
105 mL glass container, containing the granules or
spheroids to be examined, which is subjected to
horizontal oscillations. The frequency and duration
of the oscillations can be varied continuously. The
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frequency can be adjusted, using a scale, to a value
in the range 0-400 oscillations/min. The duration
can be set to a value in the range 0-9999 s.
Procedure
The following procedure is usually suitable.
Remove the fine particles by sieving (sieve having
an aperture size of 355 µm or any other suitable
sieve). In the glass container, weigh about 10.00 g
(m1) of the granules or spheroids. Install the
container in the apparatus.
Shake for 240 s at the highest frequency for hard
granules or spheroids, or for 120 s at a lower
frequency (e.g. 140 oscillations/min) for soft
granules or spheroids. Sieve (355 µm, or the same
sieve as used previously) and weigh the granules or
spheroids again (m2). Test 3 samples and calculate
the mean value.
Loss on drying Dry in an oven at 105 °C, unless
otherwise prescribed. Alternatively, other
drying conditions as described in general method
may be used.
Calculation
F = Friability;
T1 = Percentage loss on drying before the test (mean
of 2 determinations);
T2 = Percentage loss on drying after the test (mean
of 2 determinations);
m1 = Mass of the granules or spheroids before the
test, in grams;
m2 = Mass of the granules or spheroids after the test,
in grams.
Adnan Sarwar Chaudhary
Acceptance criteria:
 Not more than 1% USP
 Not more than 0.8% BP
Procedure:
 For tablets weighing upto 0.65g, take a
sample of 20 tablets.
 For tablets weighing more than 0.65g, take a
sample of 10 tablets.
 Place the tablet on a sieve no. 1000 and
remove any loose dust with the aid of air
pressure or a soft brush.
 Accurately weigh the tablet samples and
place the tablet in the drum.
 Rotate the drum 100 times (25 rpm for 4
minutes) and remove the tablets.
 Remove any loose dust from the tablets as
before. If no tablets are cracked, split or
broken, weigh the tablets to the nearest mg.
 If the results are doubtful, repeat he test 3
times, take the average, average should be
maximum to 1%.
Formula:
F=100(1- )
 100 = Revolutions
 Wo = Weight before test
 W1 = Weight after test
Disadvantages
 Affect the elegance of tablets.
 Affect the consumer acceptance of tablets
Disintegration
The state in which any residue of the unit, except
fragment of insoluble coating or capsule shell,
remaining on the screen of the test apparatus or
adhering to the lower surface of the disk, if used, is
a soft mass having no palpably firm core.
Theories of disintegration:
Several mechanisms of tablet disintegration as been
proposed. Some of them are given below:
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1. Evalution of gas:
If the gas is evolved by a chemical reaction, when
the tablet comes into contact with water, then the
tablet will disintegrate. This is the basis for
manufacture of effervescent tablets.
Example of such a reaction is sodium bicarbonate
with citric and tartaric acid, which yields carbon
dioxide.
2. Heat of wetting:
The heat produced when a tablet is immersed in
water causes the entrapped air in the tablet to
expand and exert sufficient pressure to disintegrate
the tablet.
3. Effect of water absorption:
The water absorbed by the tablet initiate
disintegration, but this depends upon the solubility
of the drug and other ingredients present.
4. Swelling:
The grains of the disintegrant, particularly of
starches, swell in the present of water and exert
pressure on the granules to force them apart.
5. Porosity of tablets:
It has been shown that penetration of water into a
tablet is proportional to its mean pore diameter or
porosity. The porosity and permeability of tablets
decrease as the tableting pressure increased, and as
the porosity decrease, the disintegration time
increases.
Purpose:
This test is performed to determine that whether the
tablets or capsules disintegrate within the prescribed
time when placed in a liquid medium under the
specified experimental conditions
Apparatus:
Types of apparatus
Two types of apparatus are used:
 Apparatus-A:
Most commonly used for tablets which are
less than 18 mm long
 Apparatus-B:
Adnan Sarwar Chaudhary
Most commonly used for tablets which are
more than 18 mm long
Apparatus construction
1. Circular basket rack assembly
2. Suitable vessel for immersion fluid (1 liter
beaker)
3. Thermostatic arrangement for maintaining
the temperature at 37+2 oC
4. A device for rising and lowering of basket
rack in immersion fluid at constant
frequency of 28-32 cycles/min through a
distance of 50-60 mm.
Liquids used in disintegration:
1. Water
2. Simulated gastric fluid (PH = 1.2)
3. Simulated intestinal fluid (PH = 7.5 )
Procedure:
Place 1 tablet in each of the six tubes of basket and
operate the apparatus using water maintained at
37±20.
At the end of time limit, lift the basket from the
fluid and observe the tablets, if all of the tablets
have disintegrated, test is clear, if 1-2 tablets fail to
disintegrate, repeat the apparatus at 12 additional
tablets.
1. Disintegration Test for Tablets
and Capsules
This test is provided to determine whether tablets or
capsules disintegrate within the prescribed time
when placed in a liquid medium under the
experimental conditions presented below.
For the purposes of this test, disintegration does not
imply complete dissolution of the unit or even of its
active constituent. Complete disintegration is
defined as that state in which any residue of the
unit, except fragments of insoluble coating or
capsule shell, remaining on the screen of the test
apparatus or adhering to the lower surface of the
discs, if used, is a soft mass having no palpably firm
core.
Use apparatus A for tablets and capsules that are not
greater than 18 mm long. For larger tablets or
capsules use apparatus B.
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Test A – Tablets and capsules of
normal size
Apparatus
The apparatus consists of a basket-rack assembly, a
1 litre, low-form beaker, 149 ± 11 mm in height and
having an inside diameter of 106 ± 9 mm for the
immersion fluid, a thermostatic arrangement for
heating the fluid between 35 °C and 39 °C, and a
device for raising and lowering the basket in the
immersion fluid at a constant frequency rate
between 29 and 32 cycles per minute, through a
distance of 55 ± 2 mm. The volume of the fluid in
the vessel is such that at the highest point of the
upward stroke the wire mesh remains at least 15
mm below the surface of the fluid, and descends to
not less than 25 mm from the bottom of the vessel
on the downward stroke. At no time should the top
of the basket-rack assembly
become submerged. The time required for the
upward stroke is equal to the time required for
the downward stroke, and the change in stroke
direction is a smooth transition, rather than an
abrupt reversal of motion. The basket-rack
assembly moves vertically along its axis. There is
no appreciable horizontal motion or movement of
the axis from the vertical.
Basket-rack assembly
The basket-rack assembly consists of 6 open-ended
transparent tubes, each 77.5 ± 2.5 mm long and
having an inside diameter of 21.85 ± 1.15 mm and a
wall 1.9 ± 0.9 mm thick; the tubes are held in a
vertical position by 2 plates, each 90 ± 2 mm in
diameter and 6.75 ± 1.75 mm in thickness, with 6
holes, each 24 ± 2 mm in diameter, equidistant from
the centre of the plate and equally spaced from one
another. Attached to the under surface of the lower
plate is a woven stainless steel wire cloth, which has
a plain square weave with 2.0 ± 0.2 mm mesh
apertures and with a wire diameter of 0.615 ± 0.045
mm. The parts of the apparatus are assembled and
rigidly held by means of 3 bolts passing
through the 2 plates. A suitable means is provided
to suspend the basket-rack assembly from the
raising and lowering device using a point on its
axis. The design of the basket-rack assembly may
Adnan Sarwar Chaudhary
be varied somewhat provided the specifications for
the glass tubes and the screen mesh size are
maintained. The basket-rack assembly conforms to
the dimensions shown in Figure 2.9.1.-1.
Discs
The use of discs is permitted only where specified
or allowed. Each tube is provided with a cylindrical
disc 9.5 ± 0.15 mm thick and 20.7 ± 0.15 mm in
diameter. The disc is made of a suitable, transparent
plastic material having a specific gravity of 1.18-
1.20. 5 parallel 2 ± 0.1 mm holes extend between
the ends of the cylinder. One of the holes is
centered on the cylindrical axis. The other holes are
centered 6 ± 0.2 mm from the axis on imaginary
lines perpendicular to the axis and parallel to each
other. 4 identical trapezoidal-shaped planes are
cut into the wall of the cylinder, nearly
perpendicular to the ends of the cylinder. The
trapezoidal shape is symmetrical; its parallel sides
coincide with the ends of the cylinder and
are parallel to an imaginary line connecting the
centers of 2 adjacent holes 6 mm from the
cylindrical axis. The parallel side of the trapezoid
on the bottom of the cylinder has a length of
1.6 ± 0.1 mm and its bottom edges lie at a depth of
1.5 mm to 1.8 mm from the cylinder's
circumference. The parallel side of the trapezoid on
the top of the cylinder has a length of 9.4
± 0.2 mm and its center lies at a depth of 2.6 ± 0.1
mm from the cylinder's circumference. All surfaces
of the disc are smooth. If the use of discs is
specified, add a disc to each tube and operate the
apparatus as directed under Procedure. The discs
conform to the dimensions shown in Figure 2.9.1.-
1. The use of automatic detection employing
modified discs is permitted where the use of discs
is specified or allowed. Such discs must comply
with the requirements of density and
dimension given in this chapter.
Procedure
Place 1 dosage unit in each of the 6 tubes of the
basket and, if prescribed, add a disc. Operate the
apparatus using the specified medium, maintained
at 37 ± 2 °C, as the immersion fluid. At the end of
the specified time, lift the basket from the fluid and
observe the dosage units: all of the dosage units
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have disintegrated completely. If 1 or 2 dosage units
fail to disintegrate, repeat the test on 12 additional
dosage units. The requirements of the test are met if
not less than 16 of the 18 dosage units tested have
disintegrated.
Test B – Large tablets and large
capsules
Apparatus
The main part of the apparatus (Figure 2.9.1.-2.) is a
rigid basket-rack assembly
supporting 3 cylindrical transparent tubes 77.5 ± 2.5
mm long, 33.0 mm ± 0.5 mm in internal
diameter, and with a wall thickness of 2.5 ± 0.5
mm. Each tube is provided with a cylindrical
disc 31.4 ± 0.13 mm in diameter and 15.3 ± 0.15
mm thick, made of transparent plastic with a
relative density of 1.18-1.20. Each disc is pierced
by 7 holes, each 3.15 ± 0.1 mm in diameter, 1 in the
centre and the other 6 spaced equally on a circle of
radius 4.2 mm from the centre of
the disc. The tubes are held vertically by 2 separate
and superimposed rigid plastic plates 97 mm in
diameter and 9 mm thick, with 3 holes. The holes
are equidistant from the centre of the plate and
equally spaced. Attached to the underside of the
lower plate is a piece of woven gauze made from
stainless steel wire 0.63 ± 0.03 mm in diameter and
having mesh apertures of 2.0 ± 0.2 mm. The plates
are held rigidly in position and 77.5 mm apart by
vertical metal rods at the periphery. A metal rod is
also fixed to the centre of the upper plate to enable
the assembly to be attached to a mechanical device
capable of raising and lowering it smoothly at
a constant frequency of between 29 and 32 cycles
per minute, through a distance of 55 ± 2 mm.
The assembly is suspended in the specified liquid
medium in a suitable vessel, preferably a 1 litre
beaker. The volume of the liquid is such that when
the assembly is in the highest position the wire
mesh is at least 15 mm below the surface of the
liquid, and when the assembly is in the lowest
position the wire mesh is at least 25 mm above the
bottom of the beaker and the upper open ends of the
tubes remain above the surface of the liquid. A
Adnan Sarwar Chaudhary
suitable device maintains the temperature of the
liquid at 35-39 °C.
The design of the basket-rack assembly may be
varied provided the specifications for the tubes and
wire mesh are maintained.
Method
Test 6 tablets or capsules either by using 2 basket-
rack assemblies in parallel or by repeating the
procedure. In each of the 3 tubes, place 1 tablet or
capsule and, if prescribed, add a disc; suspend the
assembly in the beaker containing the specified
liquid.
Operate the apparatus for the prescribed period,
withdraw the assembly and examine the state of the
tablets or capsules. To pass the test, all 6 of the
tablets or capsules must have disintegrated.
Acceptance criteria:
All the 6 tablets or capsules must be disintegrated.
If 1 or 2 dosage unit/s fails to disintegrate than
repeat the
test with additional 12 tablets. The requirements of
the test are meet if not less than 16 of the 18 dosage
units
tested are disintegrated.
Acceptance/rejection criteria:
i. Uncoated tablets:
completely, repeat the test on 12 additional tablets,
not less than 16 of total 18 tablets disintegrate
completely.
 According to USP disintegration time must
be less than 30min.
 According to BP disintegration time must be
less than 15min.
ii. Plain coated tablets (film/sugar coated):
Prescribed time and acceptance/rejection criteria are
the same as to the uncoated tablets.
iii. Enteric coated tablets:
 One tablet in each tube.
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 Operate the apparatus using simulated
gastric fluid till 1-2 hours.
 Lift the basket from fluid and observed the
tablets.
 There should be no evidence of
disintegration, cracking and softening.
 Operate the apparatus using simulated
intestinal fluid for one hour or the time
specified in the monograph.
 Acceptance/rejection criteria are same as of
the uncoated tablets.
The disintegration test determines whether the
suppositories or pessaries soften or disintegrate
within the prescribed time when placed in a liquid
medium in the experimental conditions described
below.
Disintegration is considered to be achieved when:
a) dissolution is complete,
b) the components of the suppository or pessary
have separated: melted fatty substances collect on

iv. Buccal tablets:

 One tablet in each tube.
 Operate the apparatus using water or
specified medium, if prescribed.
 After 4 hours all the tablets should have
disintegrated.
 Acceptance/rejection criteria are same as of
the uncoated tablets.

v. Sublingual tablets:
 Prescribed medium and A/R criteria is same.
 Omit the use of disc.
 Observe the tablets for time specified in
monograph.
 For the most of the sublingual tablets time
limit is within 2min.

vi. Hard gelatin capsules:

 Apply the test of uncoated tablets.
 Attach the removable wire cloth, which has
a plain square weave with 1.8 to 2.2 mm
mesh apertures and with a wire diameter of
0.60 – 0.655mm, as described under basket-
rack assembly, to the surface of the upper
plate of basket-rack assembly.
 Observe the capsules within the time limit
specified in the individual monograph.
 All the capsule have disintegrate except the
capsule shell.
vii. Soft gelatin capsules:
Proceed as describer under hard gelatin capsules.
Disintegration Test for
Suppositories and Pessaries
the surface of the liquid, insoluble powders fall to
the bottom and soluble components dissolve,
depending on the type of preparation, the
components may be distributed in one or more of
these ways,
c) there is softening of the sample that may be
accompanied by appreciable change of shape
without complete separation of the components, the
softening is such that the suppository or pessary no
longer has a solid core offering resistance to
pressure of a glass rod,
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d) rupture of the gelatin shell of rectal or vaginal
capsules occurs allowing release of the contents,
e) no residue remains on the perforated disc or if a
residue remains, it consists only of a soft or frothy
mass having no solid core offering resistance to
pressure of a glass rod (vaginal tablets).
Apparatus
The apparatus (Figure 2.9.2.-1) consists of a sleeve
of glass or suitable transparent plastic, of
appropriate thickness, to the interior of which is
attached by means of three hooks a metal device
consisting of two perforated stainless metal discs
each containing 39 holes 4 mm in diameter; the
diameter of the discs is similar to that of the
interior of the sleeve; the discs are about 30 mm
apart. The test is carried out using three such
apparatuses each containing a single sample. Each
apparatus is placed in a beaker with a capacity of at
least 4 L filled with water maintained at 36-37 °C,
unless otherwise prescribed. The apparatuses may
also be placed together in a vessel with a capacity
of at least 12 L. The beaker is fitted with a slow
stirrer and a device that will hold the cylinders
vertically not less than 90 mm below the surface of
the water and allow them to be inverted without
emerging from the water.
Method
Use three suppositories or pessaries. Place each one
on the lower disc of a device, place the latter in the
Adnan Sarwar Chaudhary
sleeve and secure. Invert the apparatuses every 10
min. Examine the samples after the period
prescribed in the monograph. To pass the test all
the samples must have disintegrated.
Method of operation for vaginal tablets
Use the apparatus described above, arranged so as
to rest on the hooks (see Figure 2.9.2.-2). Place it in
a beaker of suitable diameter containing water
maintained at 36- 37 °C with the level just below
the upper perforated disc. Using a pipette, adjust the
level with water at 36-37 °C until a uniform film
covers the perforations of the disc. Use three
vaginal tablets. Place each one on the upper plate of
an apparatus and cover the latter with a glass plate
to maintain appropriate conditions of humidity.
Examine the state of the samples after the period
prescribed in the monograph. To pass the test all
the samples must have disintegrated.,
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 Now calculate the percentage deviation by

following formulame batch for assay
determinations.]

PercentageDeviation= ×100

Uncoated or Film-Coated Tablets

Accurately weigh 10 tablets individually. Calculate
the drug substance content, expressed as % of label
claim, of each tablet from the weight of the
individual tablet and the result of the Assay.
Calculate the acceptance value.

Acceptance criteria:
Not more than 2 of the individual masses deviate
from the average mass by more than the percentage
deviation given in the following table.
Monographs of the British Pharmacopoeia According to USP
The following additional points apply to Average weight of %age deviation
monographs of the British Pharmacopoeia. tablet(mg)
130 mg or less ±10%
ACCEPTANCE CRITERIA 130-324 mg ±7.5%
For moulded suppositories, disintegration occurs in More than 324 mg ±5%
not more than 30 minutes for fat- based According to BP
suppositories and in not more than 60 minutes for
Average weight %age deviation
water-soluble suppositories, unless otherwise
80 mg or less ±10%
justified and authorised.
More than 80 to less than ±7.5%
For moulded pessaries, disintegration occurs in not
250
more than 60 minutes unless otherwise justified and More than 250 mg ±5%
authorised.
For rectal capsules and vaginal tablets and capsules,
disintegration occurs in not more than 30 minutes. Hard Capsules
Accurately weigh 10 capsules individually, taking
Weight Variation care to preserve the identity of each capsule.
Select not fewer than 30 dosage units, and proceed Remove the contents of each capsule by a suitable
as follows for the dosage form designated. The means. Accurately weigh the emptied shells
result of the Assay, obtained as directed in the individually, and calculate for each capsule the net
individual monograph, is designated as result A, weight of its contents by subtracting the weight of
expressed as % of label claim. Assume that the the shell from the respective gross weight. Calculate
concentration (weight of drug substance per weight the drug substance content, expressed as % of label
of dosage unit) is uniform. [NOTE—Specimens claim, of each capsule from the net weight of the
other than these test units may be drawn from the sa individual capsule content and the result of the
Assay. Calculate the acceptance value.
For Tablets:
 Weigh 20 tablets individually and calculate
average weight.
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Soft Capsules
Accurately weigh 10 intact capsules individually to
obtain their gross weights, taking care to preserve
the identity of each capsule. Then cut open the
capsules by means of a suitable clean, dry cutting
instrument such as scissors or a sharp open blade,
and remove the contents by washing with a suitable
solvent. Allow the occluded solvent to evaporate
from the shells at room temperature over a period of
about 30 minutes, taking precautions to avoid
uptake or loss of moisture. Weigh the individual
shells, and calculate the net contents. Calculate the
drug substance content, expressed as % of label
claim, in each capsule from the net weight of
product removed from the individual capsules and
the result of the Assay. Calculate the acceptance
value.
Acceptance criteria:
Not more than 2 of the individual masses deviate
from the average mass by more than the percentage
deviation given in the following table.
Average weight %age deviation
Less than 300 mg ±10%
More than 300 mg ±7.5%
For Powders
 Remove any paper label from outside then
dry and wash it from outside.
 Open the container and weigh the container
and its contents.
 Empty the container as completely as
possible by gentle tapping; rinse it if
necessary with water or Alcohol.
 Dry it in oven.
 Allow to cool in desiccators and weigh the
mass of contents i.e. the difference between
the weighing.
 Repeat the procedure with another 19
containers.
Acceptance Criteria:
Average weight %age deviation
More than 40mg ±10%
Adnan Sarwar Chaudhary
Importance:
This test is important in terms of
Safety of dosage unit, proper weight, proper dose
Solids (Including Sterile Solids) in
Single-Unit Containers
Proceed as directed for Hard Capsules, treating each
unit as described therein. Calculate the acceptance
value.
Oral Solutions Packaged in Single-
Unit Containers
Accurately weigh the amount of liquid that drains
in not more than 5 seconds from each of 10
individual containers. If necessary, compute the
equivalent volume after determining the density.
Calculate the drug substance content, expressed as
% of label claim, in the liquid drained from each
unit from the net weight of the individual
container content and the result of the Assay.
Calculate the acceptance value.
Calculation of Acceptance Value
Calculate the acceptance value as shown in Content
Uniformity, except that the individual contents of
the units are replaced with the individual
estimated contents defined below.
X1, X2……….., X= individual estimated contents of the
units tested,
Where
Xi= w × A / W
i
W= individual weights of the units
tested (w1, w2,……………, wn)
A= content of drug substance (% of label claim)
determined as described in the Assay, and
Solutions for Inhalation Packaged
in Glass or Plastic Ampuls and
Intended for Use in Nebulizers
[NOTE—Acceptance value
calculations are not required
for these dosage forms.]
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Accurately weigh 10 containers individually, taking
care to preserve the identity of each container.
Remove the contents of each container by a suitable
means. Accurately weigh the emptied containers
individually, and calculate for each container the net
weight of its contents by subtracting the weight of
the container from the respective gross weight.
From the results of the Assay, obtained as directed
in the individual monograph, calculate the drug
substance content, expressed as % of label claim, in
each of the containers.
CRITERIA
Apply the following criteria, unless otherwise
specified in the individual monograph.
Uncoated, Coated, or Molded Tablets, Capsules,
Oral Solutions in Single-Unit Containers, Oral
Suspensions or Oral Emulsions or Oral Gels in
Single-Unit Containers, and Solids (Including
Sterile Solids) in Single-Unit Containers
The requirements for dosage uniformity are met if
the acceptance value of the first 10 dosage units is
less than or equal to L1%. If the acceptance value
is greater than L1%, test the next 20 units and
calculate the acceptance value. The requirements
are met if the final acceptance value of the 30
dosage units is less than or equal to L1%, and no
individual content of any dosage unit is less than
(1– L2*0.01)M nor more than (1 + L2*0.01)M as
specified in the Calculation of Acceptance Value
under Content Uniformity or under Weight
Variation. Unless otherwise specified in the
individual monograph, L1 is 15.0 and L2 is 25.0.
Suppositories
Limit A (if the average of the limits specified in the
potency definition in the individual monograph is
100.0 percent or less)
Unless otherwise specified in the individual
monograph, the requirements for dosage uniformity
are met if the amount of the drug substance in each
of the 10 dosage units as determined from the
Content Uniformity method lies within the range of
85.0% to 115.0% of the label claim, and the RSD is
less than or equal to 6.0%.
Adnan Sarwar Chaudhary
If 1 unit is outside the range of 85.0% to 115.0% of
label claim, and no unit is outside the range of
75.0% to 125.0% of label claim, or if the RSD is
greater than 6.0%, or if both conditions prevail,
test 20 additional units. The requirements are met if
not more than 1 unit of the 30 is outside the
range of 85.0% to 115.0% of label claim, and no
unit is outside the range of 75.0% to 125.0% of
label claim and the RSD of the 30 dosage units does
not exceed 7.8%.
Limit B (if the average of the limits specified in the
potency definition in the individual monograph is
greater than 100.0 percent)
1. If the average value of the dosage units tested is
100.0 percent or less, the requirements are as in
Limit A.
2. If the average value of the dosage units tested is
greater than or equal to the average of the limits
specified in the potency definition in the individual
monograph, the requirements are as specified under
Limit A, except that the words ―label claim‖ are
replaced by the words ―label claim multiplied by the
average of the limits specified in the potency
definition in the monograph divided by 100‖.
3. If the average value of the dosage units tested is
between 100 percent and the average of the limits
specified in the potency definition in the individual
monograph, the requirements are as specified under
Limit A, except that the words ―label claim‖ are
replaced by the words ―label claim multiplied by the
average value of the dosage units tested (expressed
as a percent of label claim) divided by 100‖.
Transdermal Systems and Inhalations Packaged
in Premetered Dosage Units
Limit A (if the average of the limits specified in the
potency definition in the individual monograph is
100.0 percent or less)
Unless otherwise specified in the individual
monograph, the requirements for dosage uniformity
are met if the amount of the drug substance in not
fewer than 9 of the 10 dosage units as determined
from the Content Uniformity method (or, in the case
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of solutions for inhalation packaged in glass or
plastic ampuls and intended for use in nebulizers,
from either the Content Uniformity or the Weight
Variation method) lies within the range of 85.0% to
115.0% of label claim, and no unit is outside the
range of 75.0% to 125.0% of label claim, and the
RSD of the 10 dosage units is less than or equal to
6.0%.
If 2 or 3 dosage units are outside the range of 85.0%
to 115.0% of label claim, but not outside the
range of 75.0% to 125.0% of label claim, or if the
RSD is greater than 6.0% or if both conditions
prevail, test 20 additional units. The requirements
are met if not more than 3 units of the 30 are
outside the range of 85.0% to 115.0% of label claim
and no unit is outside the range of 75.0% to
125.0% of label claim, and the RSD of the 30
dosage units does not exceed 7.8%.
Limit B (if the average of the limits specified in the
potency definition in the individual monograph is
greater than 100.0 percent)
1. If the average value of the dosage units tested is
100.0 percent or less, the requirements
are as in Limit A.
2. If the average value of the dosage units tested is
greater than or equal to the average of
the limits specified in the potency definition in the
individual monograph, the requirements
are as specified under Limit A, except that the
words ―label claim‖ are replaced by the
words ―label claim multiplied by the average of the
limits specified in the potency definition
in the monograph divided by 100‖.
3. If the average value of the dosage units tested is
between 100 percent and the average of the limits
specified in the potency definition in the individual
monograph, the requirements are as specified under
Limit A, except that the words ―label claim‖ are
replaced by the words ―label claim multiplied by the
average value of the dosage units tested (expressed
as a percent of label claim) divided by 100‖).
Chemical Tests
Adnan Sarwar Chaudhary
Content uniformity
This test is used to confirm that every unit should
contain same amount of drug or active ingredients
with little variation in the batch.
Principle
The test of the uniformity of the content of single
dose preparations is based on the assay of individual
contents of active substance(s) of a number of
dosage units to determine whether the individual
contents are within the limits set with reference to
the average content of the sample.
Method
 Using a suitable analytical method,
determine the individual contents of active
substance(s) of 10 dosage units taken at
random.
 Now apply the criteria of Test A, Test B or
Test C as specified in the monograph for any
specified dosage form.
Test A:
 For tablets, powders for parenteral use,
ophthalmic inserts suspension for injection.
 The preparation complies with the test if
each individual content is between 85-115%
of the Average content.
 The preparation fails to comply with the test
if more than 1 individual content is outside
85-115% of the average content or if 1
individual content is outside 75-125% of the
average content.
 If 1 individual content is outside the limit of
85-115% but within the limit of 75-125%,
determine the individual content of another
20 dosage units taken at random.
 The preparation complies with the test if
more than 1 individual content of the 30
units is outside 85-115% of the average
content and none is outside the limit of 75-
125% of the average content.
Test B:
 For capsules, powders other than parenteral
use, granules, suppositories, pessaries.
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 Complies with test if not more than 1
individual content is outside 85-115% of the
average content and none is outside the
limit of 75-125% of the average content.
 Fails to comply with test if more than 3
individual contents are outside 85-115% of
the average content or if 1 or more
individual contents are outside the limit of
75-125% of the average content.
 If 2 or 3 individual are outside the limit of
85-115% than take 20 more units and
perform as in Test A.
Test C:
 For transdermal patches.
 Complies with test if the average content of
the 10 dosage units is between the limit of
90- 110% of the content stated on the label
and if the individual content of each dosage
unit is between the limit of 75-125% of the
average content.
DISSOLUTION TEST (USP)
It is the amount of drug that is dissolved or goes
into the solution per unit time under the specified
conditions.
Test Specification
Dissolution medium:
Test is begin with aqueous media of pH 1.2-6.8.
Simple water is usually not recommended due to the
ionic strength and pH can be vary. Different type of
dissolution medium are as follow:
i. Water or medium pH less than 6.8 with
addition of purified pepsin (Intestinal
environment)
ii. Medium with pH 6.8 or greater or pancreatic
medium can be added (protease activity)
iii. 0.1N HCl with pH 1.2
iv. Acetate buffer with pH 4.5
v. Phosphate buffer with pH 6.8-7.5
vi. Delayed release dosage form pH 6.8
Adnan Sarwar Chaudhary
Temperature:
Temperature is maintained according to body
temperature i.e. 37 ± 2o C.
Speed of the apparatus:
50-100 rpm
Sink condition:
Concentration that yield saturation solubility of
drug substances at least three time the weight dose
of the drug substance dissolved in the volume of
the medium used for dissolution.
Example
If the dissolution of 100mg strength tablet is being
performed in 900ml of the medium. A saturation
solubility is greater than 0.33mg/ml in the medium
is required to maintain sink condition.
Reference Standard USP
Non-disintegrated USP Chlorpheniramine Maleate
Extended-Release Tablets (Apparatus III)
USP Salicylic Acid Tablets (Apparatus I & II)
Disintegrated
USP Prednisone Tablets (Apparatus I & II)
Dissolution Test
 In 1970‘s dissolution test for dissolution
introduce first time.
 In 1970‘s USP recommended 7 apparatus
for dissolution test
 In 1975‘s BP recommended 4 apparatus for
dissolution test
Dissolution Apparatus
According to USP 7 types of apparatus are used
which are as follow:
1. Apparatus I (Rotating Basket Apparatus)
2. Apparatus II (Rotating Paddle Apparatus)
3. Apparatus III (Reciprocating Cylinder
Apparatus)
4. Apparatus IV (Flow Through Cell
Apparatus)
5. Apparatus V (Paddle Over Disc Apparatus)
6. Apparatus VI (Rotating Cylinder Apparatus)
7. Apparatus VII (Reciprocating Holder
Apparatus)
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Apparatus I (Rotating Basket capacity of 4 L, the height is 280 mm to 300 mm

and its inside diameter is 145 mm to 155 mm.
Apparatus)
 The shaft is positioned so that its axis is not
Assembly: more than 2 mm at any point from the
Vessel vertical axis of the vessel and rotates
May be covered (to retard evaporation), made of smoothly and without significant wobble
glass or other inert, transparent material of 100ml that could affect the results.
capacity containing 900ml of dissolution medium.  A speed-regulating device is used that
It is of cylindrical shape with hemispherical ends. allows the shaft rotation speed to be selected
and maintained at the specified rate given in
Motor or Shaft the individual monograph, within ±4%.
A metallic drive shaft for rotation  Shaft and basket components of the stirring
element are fabricated of stainless steel or
Cylindrical basket
other inert material.
Place dosage form on it having 40 mesh.
 A basket having a gold coating of about
Temperature regulator or water bath 0.0001 inch (2.5 µm) thick may be used.
Maintain the temperature of medium at 37 ± 0.5 oC  A dosage unit is placed in a dry basket at the
beginning of each test.
Fitted cover
 The distance between the inside bottom of
Use to retard the evaporation of dissolution medium
the vessel and the bottom of the basket is
Working maintained at 25 ± 2 mm during the test.
 The vessel is partially immersed in a
suitable water bath of any convenient size or
heated by a suitable device such as a heating
jacket.
 The water bath or heating device permits
holding the temperature inside the vessel at
37 ± 0.5 during the test and keeping the
bath fluid in constant, smooth motion.
 No part of the assembly, including the
environment in which the assembly is
placed, contributes significant motion,
agitation, or vibration beyond that due to the
smoothly rotating stirring element.
 An apparatus that permits observation of the
specimen and stirring element during the test
is preferable.
Uses:
The vessel is cylindrical, with a hemispherical
 Generally preferred for capsules
bottom and with one of the following dimensions
 Dosage form that tend to dissolved slowly
andcapacities: for a nominal capacity of 1 L, the
height is 160 mm to 210 mm and its inside diameter
is 98 mm to 106 mm; for a nominal capacity of 2 L,
the height is 280 mm to 300 mm and its inside
diameter is 98 mm to 106 mm; and for a nominal

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Apparatus II (Rotating Paddle
Apparatus)
 Use the assembly from Apparatus I, except
that a paddle formed from a blade and a
shaft is used as the stirring element.
 The shaft is positioned so that its axis is not
more than 2 mm from the vertical axis of the
vessel at any point and rotates smoothly
without significant wobble that could affect
the results.
 The paddle blade and shaft may be coated
with a suitable coating so as to make them
inert.
 The dosage unit is allowed to sink to the
bottom of the vessel before rotation of the
blade is started.
 A small, loose piece of nonreactive material,
such as not more than a few turns of wire
helix, may be attached to dosage units that
would otherwise float.
 Rotation of blade is 50rpm
Apparatus III (Reciprocating
Cylinder Apparatus)
Assembly
 Glass vessels
 Cylindrical and flat-bottomed
 Glass reciprocating cylinders
Adnan Sarwar Chaudhary
 Inert fittings (stainless steel type 316 or
other suitable material)
 Mesh Screens
 They are made of suitable nonsorbing and
nonreactive material and that are designed to
fit the tops and bottoms of the reciprocating
cylinders
 Motor and drive assembly
 To reciprocate the cylinders vertically inside
the vessels and, if desired, index the
reciprocating cylinders horizontally to a
different row of vessels.
Working:
 The vessels are partially immersed in a
suitable water bath of any convenient size
that permits holding the temperature at 37 ±
0.5 during the test.
 Other parts of working are same as in
Apparatus I but instead of rpm, dpm (dips
per minute) are used.
5dpm = 50rpm
Uses
 Chewable tablets
 Conventional dosage form
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Apparatus IV (Flow Through Cell

Apparatus)
Assembly:
 Reservoir and a pump for the Dissolution
Medium
 A flow-through cell
 A water bath that maintains the Dissolution
Medium at 37 ± 0.5.
 Use the specified cell size as given in the
individual monograph. Apparatus V (Paddle Over Disc
Working: Apparatus)
 Modified form of Apparatus II
Pump
 It is use to reduce the dead space (space
 The pump forces the Dissolution Medium
between paddle and dosage unit)
upwards through the flow-through cell.
 Use for individual monograph specifications
 The pump has a delivery range between 240
and 960 mL per hour, with standard flow Working:
rates of 4, 8, and 16 mL per minute.  Place the dosage form on disc (place cellular
membrane for transdermal patches)
Flow rate
 Adjust the space 25 ± 2 mm between paddle
 It must deliver a constant flow (±5% of the
and disc.
nominal flow rate); the flow profile is
sinusoidal with a pulsation of 120 ± 10  Volume of medium used is 1000 ml.
pulses per minute.  Maintain the temperature at 32 ± 0.5 oC.

Flow through cell & material Uses:

The flow-through cell, of transparent and inert 1. Transdermal patches
material, is mounted vertically with a filter system 2. Sustained release dosage form
(specified in the individual monograph) that 3. Modified or extended release dosage form
prevents escape of undissolved particles from the
top of the cell; standard cell diameters are 12 and
22.6 mm; the bottom cone is usually filled with
small glass beads of about 1-mm diameter with one
bead of about 5 mm positioned at the apex to
protect the fluid entry tube; and a tablet holder is
available for positioning of special dosage forms,
for example, inlay tablets.

Temperature maintenance
The cell is immersed in a water bath, and the
temperature is maintained at 37 ± 0.5 oC.
Apparatus VI (Rotating Cylinder
Apparatus)
 Modified form of Apparatus I
 Basket is replaced with stainless steel
cylinder.
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 Transdermal patch is attached with cellular the stomach

membrane which is than attached with Absorption is primary The rate of dissolution is
focus in drug‘s the key target for
cylinder.
development controlling drug duration
 Maintain the temperature at 32 ± 0.5 oC. and medicinal chemistry in vive.
since the drug much be
Uses: absorbed before any effect
1. Transdermal patches can take place.
No equation for the Noye‘s Whitney equation
Apparatus VII (Reciprocating measurement of is used for measuring
absorption. the rate of dissolution.
Holder Apparatus) For the absorption the drug Dissolution of the drug in
much be soluble in the equeous medium
 A set of volumetrically calibrated or tared water. depends upon the pH of
solution container made up of glass or any medium
other material
 A holder to hold the dosage unit Assay of active Ingredient
 Reciprocating frequency is about 30 cycles
per minute.
Definition
 Motor and shaft drive assembly.
 The determination of activity, potency, strength,
Uses: etc of a substance, either on absolute basis or in
1. Transdermal patches comparison with that of standard preparation.
2. Extented release drug products  Qualitative or quantitative analysis of a
substance, especially of the drug, to determine
Stage Number Acceptance Criteria its components.
Tested
S1 6 Each unit is not less than Q + 5%. Specification of assay determination:
S2 6 Average of 12 units (S1 + S2) is equal 1. Temperature range = 15 and 250c.
to or greater than Q, and no unit is
less than Q 15%. 2. Carried out in diffuse light.
S3 12 Average of 24 units (S1 + S2 + S3) is 3. Percentage limit is ± 15%
equal tour greater than Q, not more
than 2 units are less than Q 15%, and
no unit is less than Q 25%.

Applications:
Qualities of standard:
1. To evaluate the formulation effect on the
1. It is a representative, selected sample of any
oral absorption of poorly water soluble given substance
drugs using a dissolution system. 2. Used to determine the potency and relative
2. To provide the criteria in-vitro drug release efficacy.
information for both the quality control 3. It should be uniform in quality and pure as
purpose, i.e., to assess batch to batch possible.
consistency of solid oral dosage forms such
Difference between Chemical and Biological
as tablets, and drug development, i.e., to
Assay
predict in vivo drug release profile.
Chemical Assay Biological Assay
Difference between Absorbance and Dissolution: Less time consuming More time consuming
Absorbance Dissolution Economical Expensive
It is a movement of drug It is a situation, a tablet is More precise and accurate Less precise and accurate
into the blood stream ingested and pass Determine the amount of Measure the actual
through the esophagus to specific biological activity of a

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compound or structure given sample

moiety present in a
given sample.
Less chance of error More chance of error
Less reliable More reliable

Appar Name Assembly Critical parameters Dosage form

atus tested
1 Reciprocating Covered glass vessel, Rotating speed Capsules, Chewable
basket a motor and metallic 100rpm±4% tablets, Controlled
drive shaft, release dosage form
and a cylinder,
2 Rotating Vessel, Rotating speed 50rpm Tablets, suspension
paddle paddle form of shaft and
blade, stirring element
3 Reciprocating Flat bottom glass vessel, Dip rate 3dpm±5% Chewable tablets,
cylinder a set of glass Conventional dosage
reciprocating cylinders, form
a device ;control rate
4 Flow through A reservoir, Medium flow rate 4- Powder, Granules,
cell a pump, 16ml/min±5% Implants, Poor
flow through cell, water soluble
bath Drugs,Micropaticles,
5 Paddle over Same as apparatus 2, but Time specified in Transdermal
disk stainless steel disk is individual monograph fromulation
fitted at bottom
6 Rotating Same as apparatus 1, Time specified in Transdermal
cylinder except basket and shaft individual monograph fromulation
replaced with stainless
steel cylinder.
7 Reciprocating A motor and drive Reciprocating Transdermal
holder assembly, frequency 30 cycle formulation,
a set of solution glass /min Controlled release
containers,
a set of suitable holders.

Apparatus Dosage Unit Dosage Form Tested Advantages Disadvantages

Placement
1 Dry basket Immediate, extended pH can be changed, Limited volume,
and delayed release automatic degassing, dead zone,
dosage form. disintegration
dissolution interaction
2 Vessel s Immediate, extended, Easy use, pH change Limited volume,
and delayed release possible, automated sticking, floating,
dosage forms. sinker require.
3 Cylinder Bead type Easy pH change, Small volume i.e.
formulations. hydrodynamics can be 250ml, limited data,
directly influenced little experience.
using dip rate.

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4 Cell Having limited Easy to change media Duration necessary,

solubility. pH, sink conditions high media volume.
can be maintained. Labor, intensive.
5 Disk Transdermal patches. Apparatus 2 can be Disk restrict patch
used by adding disk. size.
6 Cylinder Transdermal patches. Higher release rate as Slow release rate as
compare to App 7. compare to App 5.
7 Holder Transdermal parches No significant motion,
and non agitation, vibration
disintegrating oral require.
dosage forms.

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QUALITY CONTROL
OF SYRUPS, ELIXIRS,
AND DISPERSE
SYSTEM

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“Liquid preparations for oral use are usually

solutions, emulsions or suspensions containing one
or more active ingredients in a suitable vehicle;
they may in some cases consist simply of a liquid
active ingredient used as such”. (WHO)

 Syrups
 Suspensions pH meter:
 Emulsions
First of all pH meter is calibrated with buffers of
 Elixirs
known pH i.e. of pH 4 & pH 10.

GENERAL TESTS FOR Then dip the electrode of meter into dosage form
and pH of solution appear on screen of instrument.
ORAL LIQUID DOSAGE
Special consideration:
FORM  pH of formulation is determined at each step
of manufacturing.
1. Physical appearance
 Syrup: pH determined by just dipping
2. PH determination
electrode into syrup.
3. Assay & content uniformity
 Suspension: first centrifuge it so that solid
4. Surface tension & Applications
separate out and now dip electrode into it.
5. Viscosity & applications
 Emulsion: first shake it then determine the
PHYSICAL APPEARANCE pH.

 It should be physically & chemically Limit:

homogenous. ± 0.05 pH variation can be occur due to more than 1
 Free from particle & microbial growth. time use of calibration curve
 Free from cloudiness and precipitation
 No phase separation present.
 No crystal growth occur in dosage form
 Free from color variation.

pH DETERMINATION
pH maintenance is very important for stability of
active ingredient.

Method of pH determination:

i. pH paper
ii. pH Meter

pH paper:
pH paper normally dips into liquid dosage form and
change in color is examined.

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Viscosity its determination Viscosity in different dosage form:

Syrups:
and application in the They having a specific value of viscosity if is
change due to degradation of formulation then by
Quality Control of measuring viscosity we can claim its formulation.

Suspensions:
Pharmaceuticals Due to flocculation the viscosity can be change.
Viscosity is a measure of the resistance of a fluid to
Emulsion:
deformation under shear stress
Due to coalescence the viscosity can be change.
The dynamic viscosity or viscosity coefficient  is
the tangential force per unit surface, known as Method of determination of
shearing stress  and expressed in pascals, viscosity:
necessary to move, parallel to the sliding plane, a Four basic methods are below but other instruments
layer of liquid of 1 square metre at a rate (v) of 1 are also present but they are modification of these
metre per second relative to a parallel layer at a four instruments:
distance (x) of 1 metre.
i. Ostwald viscometer
The ratio dv/dx is a speed gradient giving the rate of ii. Falling sphere viscometer
shear D expressed in reciprocal seconds (s-1), so iii. Cup and bob viscometer
that  = /D. iv. Cone & plate viscometer

The unit of dynamic viscosity is the pascal second Method I

(Pa·s). The most commonly used submultiple is the
millipascal second (mPa·s). Apparatus
The apparatus consists of a glass U-tube viscometer
The kinematic viscosity v, expressed in square
(Fig.5H–1) made of clear borosilicate glass and
metres per second, is obtained by dividing the
constructed in accordance with the dimensions
dynamic viscosity h by the density r expressed in
shown in the figure and in Table 5H–1. The
kilograms per cubic metre, of the liquid measured
monograph states the size of viscometer to be used.
at the same temperature, i.e. v = h/r. The kinematic
viscosity is usually expressed in square millimetres
per second.

A capillary viscometer may be used for determining

the viscosity of Newtonian liquids and a rotating
viscometer for determining the viscosity of
Newtonian and non- Newtonian liquids. Other
viscometers may be used provided that the accuracy
and precision is not less than that obtained with the
viscometers described below.

Pharmaceutical Importance:
 Predict flow properties of material
 How to handle the material
 Predict the pourability of oral dosage form
 Predict the Stability of dosage form
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The determination of viscosity using a suitable

capillary viscometer is carried out at a temperature
of 20 ± 0.1 °C, unless otherwise prescribed. The
time required for the level of the liquid to drop from
one mark to the other is measured with a stop-watch
to the nearest one-fifth of a second. The result is
valid only if two consecutive readings do not differ
by more than 1 per cent. The average of not fewer
than three readings gives the flow time of the liquid
to be examined.
Calculate the dynamic viscosity  (2.2.8) in
millipascal seconds using the formula:
Method
Fill the viscometer with the liquid being examined k = constant of the viscometer, expressed in square
through tube L to slightly above the mark G, using millimetres per second squared,
a long pipette to minimise wetting the tube above  = density of the liquid to be examined expressed
the mark. Place the tube vertically in a water bath in milligrams per cubic millimetre, obtained by
and when it has attained the specified temperature, multiplying its relative density ( ) by 0.9982,
adjust the volume of the liquid so that the bottom of t = flow time, in seconds, of the liquid to be
the meniscus settles at the mark G. Adjust the examined.
liquid to a point about 5 mm above the mark E. The constant k is determined using a suitable
After releasing pressure or suction, measure the viscometer calibration liquid.
time taken for the bottom of the meniscus to fall To calculate the kinematic viscosity (mm2·s-1), use
from the top edge of mark E to the top edge of mark the following formula: v = kt.
F. The determination may be carried out with an
Calculate, as required, either the kinematic viscosity apparatus (Figure 2.2.9.-1) having the pacifications
() in square millimetres per second (mm2 s–1) from described in Table 2.2.9.-1
the expression:
 = Kt,
or the dynamic viscosity () in millipascal seconds
(mPa s) from the expression:
 = Kt,
where \
t = time in seconds for the meniscus to fall from E
to F,
 = mass/volume (g cm–3) obtained by multiplying
the relative density, Appendix V G, of the fluid
being examined by 0.9982.
The constant (K) of the instrument is determined
using the appropriate European Pharmacopoeia
reference liquid for viscometers.
Method II (Capillary viscometer
method)

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The minimum flow time should be 350 s for size
no. 1 and 200 s for all other sizes.
Method
Fill the viscometer through tube (L) with a
sufficient quantity of the liquid to be examined,
previously brought to 20 °C unless otherwise
prescribed, to fill bulb (A) but ensuring that the
level of liquid in bulb (B) is below the exit to
ventilation tube (M). Immerse the viscometer in the
bath of water at 20 ± 0.1 °C, unless otherwise
prescribed, maintain it in the upright position and
allow to stand for not less than 30 min to allow the
temperature to reach equilibrium. Close tube (M)
and raise the level of the liquid in tube (N) up to a
level about 8 mm above mark (E). Keep the liquid
at this level by closing tube (N) and opening tube
(M). Open tube (N) and measure, with a stop-watch
to the nearest one-fifth of a second, the time
required for the level of the liquid to drop from
mark (E) to (F).
Method III (Rotating viscometer
method)
The principle of the method is to measure the force
acting on a rotor (torque) when it rotates at a
constant angular velocity (rotational speed) in a
liquid. Rotating viscometers are used for measuring
the viscosity of Newtonian (shear-independent
viscosity) or non-Newtonian liquids (shear
dependent viscosity or apparent viscosity).
Rotating viscometers can be divided in 2 groups,
namely absolute and relative viscometers. In
absolute viscometers the flow in the measuring
Adnan Sarwar Chaudhary
geometry is well defined. The measurements result
in absolute viscosity values, which can be
compared with any other absolute values. In relative
viscometers the flow in the measuring geometry is
not defined. The measurements result in relative
viscosity values, which cannot be compared with
absolute values or other relative values if not
determined by the same relative viscometer method.
Different measuring systems are available for given
viscosity ranges as well as several rotational
speeds.
Apparatus
The following types of instruments are most
common.
Concentric cylinder viscometers (absolute
viscometers)
In the concentric cylinder viscometer (coaxial
double cylinder viscometer or simply coaxial
cylinder viscometer), the viscosity is determined by
placing the liquid in the gap between the inner
cylinder and the outer cylinder. Viscosity
measurement can be performed by rotating the
inner cylinder (Searle type viscometer) or the outer
cylinder (Couette type viscometer), as shown in
Figures 2.2.10.-1 and 2.2.10.-2, respectively. For
laminar flow, the viscosity (or apparent viscosity) 
expressed in pascal-seconds is given by the
following formula:
M = torque in newton-metres acting on the cylinder
surface,
 = angular velocity in radians per second,
h = height of immersion in metres of the inner
cylinder in the liquid medium,
Ri = radius in metres of the inner cylinder,
Ro = radius in metres of the outer cylinder,
k = constant of the apparatus, expressed in radians
per cubic metre.
For non-Newtonian liquids it is indispensable to
specify the shear stress () or the shear rate () at
which the viscosity is measured. Under narrow gap
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conditions (conditions satisfied in absolute
viscometers), there is a proportional relationship
between M and  and also between  and :
where A and B are constants for the instrument and
are calculated from the following expressions:
— for concentric surface:
— for cone-plates:
M = torque in Newton-metres acting on the cone or
cylinder surface,
 = angular velocity in radians per second,
Ri = radius in metres of the inner cylinder,
R0 = radius in metres of the outer cylinder,
R = radius in metres of the cone,
h = height of immersion in metres of the inner
cylinder in the liquid medium,
 = angle in radians between the flat disk and the
cone,
 = shear stress in pascals (Pa),
 = shear rate in reciprocal seconds (s-1)
Adnan Sarwar Chaudhary
Cone-plate viscometers (absolute viscometers)
In the cone-plate viscometer, the liquid is
introduced into the gap between a flat disc and a
cone forming a define angle. Viscosity
measurement can be performed by rotating the
cone or the flat disc, as shown in Figures 2.2.10.-3
and 2.2.10.-4, respectively. For laminar flow, the
viscosity (or apparent viscosity)  expressed in
pascal-seconds is given by the following formula:
M = torque in Newton-metres acting on the flat disc
or cone surface,
 = angular velocity in radians per second,
 = angle in radians between the flat disc and the
cone,
R = radius in metres of the cone,
k = constant of the apparatus, expressed in radians
per cubic metre.
Constants A, B of the apparatus (see under
concentric cylinder viscometers).
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Spindle viscometers (relative viscometers)
In the spindle viscometer, the viscosity is
determined by rotating a spindle (for example,
cylinder- or disc-shaped, as shown in Figures
2.2.10.-5 and 2.2.10.-6, respectively) immersed in
the liquid. Relative values of viscosity (or apparent
viscosity) can be directly calculated using
conversion factors from the scale reading at a given
rotational speed.
Adnan Sarwar Chaudhary
In a general way, the constant k of the apparatus
may be determined at various speeds of rotation
using a certified viscometer calibration liquid. The
viscosity  then corresponds to the formula:
Method
Measure the viscosity (or apparent viscosity)
according to the instructions for the operation of
the rotating viscometer. The temperature for
measuring the viscosity is indicated in the
monograph. For non-Newtonian systems, the
monograph indicates the type of viscometer to be
used and if absolute viscometers are used the
angular velocity or the shear rate at which the
measurement is made. If it is impossible to obtain
the indicated shear rate exactly, use a shear rate
slightly higher and a shear rate slightly lower and
interpolate.
With relative viscometers the shear rate is not the
same throughout the sample and therefore it cannot
be defined. Under these conditions, the viscosity of
non- Newtonian liquids determined from the
previous formula has a relative character, which
depends on the type of spindle and the angular
velocity as well as the dimensions of the sample
container (Ø = minimum 80 mm) and the depth of
immersion of the spindle. The values obtained are
comparable only if the method is carried out under
experimental conditions that are rigorously the
same.
Method IV (Falling ball viscometer
method)
The determination of dynamic viscosity of
Newtonian liquids using a suitable falling ball
viscometer is performed at 20 ± 0.1 °C, unless
otherwise prescribed in the monograph. The time
required for a test ball to fall in the liquid to be
examined from one ring mark to the other is
determined. If no stricter limit is defined for the
equipment used the result is valid only if 2
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consecutive measures do not differ by more than 1.5
per cent.
Apparatus
The falling ball viscometer consists of: a glass tube
enclosed in a mantle, which allows precise control
of temperature; six balls made of glass, nickel- iron
or steel with different densities and diameters. The
tube is fixed in such a way that the axis is inclined
by 10 ± 1° with regard to the vertical. The tube has
2 ring marks which define the distance the ball has
to roll. Commercially available apparatus is
supplied with tables giving the constants, the
density of the balls and the suitability of the
different balls for the expected range of viscosity.
Method
Fill the clean, dry tube of the viscometer, previously
brought to 20 ± 0.1 °C, with the liquid to be
examined, avoiding bubbles. Add the ball suitable
for the range of viscosity of the liquid so as to
obtain a falling time not less than 30 s. Close the
tube and maintain the solution at 20 ± 0.1 °C for at
least 15 min. Let the ball run through the liquid
between the 2 ring marks once without
measurement. Let it run again and measure with a
stop-watch, to the nearest one-fifth of a second, the
time required for the ball to roll from the upper to
the lower ring mark. Repeat the test run at least 3
times.
Calculate the dynamic viscosity  in millipascal
seconds using the formula:
k = constant, expressed in millimeter squared per
second squared,
1 = density of the ball used, expressed in grams per
cubic centimetre,
2 = density of the liquid to be examined, expressed
in grams per cubic centimetre, obtained by
multiplying its relative density by 0.9982,
t = falling time of the ball, in seconds.
Adnan Sarwar Chaudhary
SURFACE TENSION
“It is defined as force per unit length which must be
applied to surface so as to counter balance the net
inward pull”.
OR
“The force required to break the intermolecular
attraction”
Units:
Dynes/cm--------------------------------in CGS system
N/m-------------------------------------------in SI
Methods for measurement:
Choice of particular method often depends on
whether surface or interfacial tension is to be
determined the accuracy and convenience desired
size of sample available and whether the effect of
time on surface tension is to be studied.
Four methods are used for determination of surface
tension:
i. Capillary rise Method:
Capillary tube is placed in liquid the surface of
capillary the liquid will raise inside the capillary
tube and meniscus is formed which may be concave
or convex due to two type of liquid
a) Wetting liquid (concave)
b) Non-wetting liquid (convex)
a) Wetting liquid:
Liquid e.g. water rises in capillary and then form
the concave meniscus which form due to wetting
ability of liquid to glass and from diagram we
calculate ―h‖ and ―r‖.
b) Non-wetting liquid
Liquid e.g. mercury don‘t wet the capillary and
form the convex meniscus at lower end of capillary
and we note the ―h‖ this method is not use.
Apply formula
y=1/2(drhg)
r = radius of capillary h = height of liquid
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d = density of liquid g = gravitational constant.
ii. Ring detachment method:
It is used to measure both the surface tension
(placed on surface) & interfacial tension (dip in
sample)
Assembly:
 A tensiometer (ring and balance attached) is
used which consists of:
 A hanging platinum iridium ring of defined
geometry
 A micro-balance,
Principle:
 Sample is poured in beaker and brought in
contact to platinum iridium ring, as ring should
be light in weight to avoid settlement.
 For measurement of surface tension, the ring is
pulled away from the surface of liquid
 The force required to detach the platinum ring
from surface is proportional to surface tension.
 Method is applicable for liquid in which ring
can be dipped, as non-wet able and sticky
liquids cannot give true surface tension,
procedure can be carried out on a controlled
temperature.
y=K.F
 K = proportionality constant that depends on
geometry of ring
 F = surface tension force
iii. Wilhelmy plate method:
Similar to ring method; instead of ring it uses a thin
plate.
Adnan Sarwar Chaudhary
Assembly:
A tensiometer (plate and balance is attached) is used
which consists of
 A thin plate of mica, glass or platinum
 A suitable balance
Mica is a group of minerals, can with stand high
temperature and inert.
Principle:
 Thin plate is place vertically so that its lower
edge nearly touches the surface of liquid
 The plate is cleaned thoroughly and attached
to a scale or balance via a thin metal wire.
 The force on the plate due to wetting is
measured with microbalance and used to
calculate the
 surface tension by following formula.
γ=
L = is the wetted parameter (constant)
iv. Drop weight method:
Assembly:
A. Capillary tube (attached to liquid reservoir
whose surface tension is to be measured).
B. Liquid drops into a tarred weighing bottle,
placed in bottom (C).
C. Bottom (tensiometer or scale)
D. Water bath (whole apparatus is immersed in
this constant temperature water bath)
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Principle:
 Take a drop of liquid whose viscosity is to
be measured on the top of capillary tube A.
 Slight vacuum is applied until a droplet from
capillary tube A reaches full size and drops
off by itself.
 This process is repeated 5 times and average
weight & volume of single drop is
calculated.
 Surface tension is calculated by following
formula.
γ=( )F
 mg = weight of liquid droplet
 r = capillary radius
 F=correlation factor.
ASSAY & CONTENT
UNIFORMITY
This test is used to confirm that every unit should
contains same amount of drug or active ingredients
with little variation in the batch.
Principle
The test of the uniformity of the content of single
dose preparations is based on the assay of individual
contents of active substance(s) of a number of
dosage units to determine whether the individual
contents are within the limits set with reference to
the average content of the sample.
Method
 Using a suitable analytical method,
determine the individual contents of active
Adnan Sarwar Chaudhary
substance(s) of 10 dosage units taken at
random.
 Now apply the criteria of Test A, Test B or
Test C as specified in the monograph for any
specified dosage form.
 Use Test A for liquid dosage form given
below:
Test A:
 For tablets, powders for parenteral use,
ophthalmic inserts, suspension for injection.
 The preparation complies with the test if each
individual content is between 85-115% of the
Average content.
 The preparation fails to comply with the test if
more than 1 individual content is outside 85-
115% of the average content or if 1 individual
content is outside 75-125% of the average
content.
 If 1 individual content is outside the limit of 85-
115% but within the limit of 75-125%,
determine the individual content of another 20
dosage units taken at random.
 The preparation complies with the test if more
than 1 individual contents of the 30 units is
outside 85-115% of the average content and
none is outside the limit of 75-125% of the
average content.
GENERAL ORAL LIQUID
DOSAGE FORM QC
TESTS
 For Syrups
 For Elixir
 For Emulsion
 For Suspension
1. FOR SYRUPS:
i. Refractive index:
Refractrometer is used to determine the refractive
index of syrup, in which quality of material is
check.
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Limits: 1.4608—1.4630
ii. Optical rotation:
Polarimeter is used to check the optical rotation of
sugar, which in term tells about the inversion of
sugar.
Limits: syrup should have less than 56° and more
than 50° optical rotation.
2. FOR ELIXIR
Alcohol Content Determination
As elixir contain alcohol 15 to 50%.
I. Distillation Method:
Used for determination of alcohol, unless otherwise
specified in the individual monograph, it is suitable
for examining mostly for
 Fluid extracts
 Tinctures
Procedure:
a) For liquids presumed to contain 30% of alcohol
or less:
 By using pipette transfer liquid dosage form
in distilling apparatus equal to 25ml.
 Note temperature and add equal volume of
water
 Pour into distillation apparatus and start the
process
 Collect the volume of distillate about 2ml
less than volume of original test liquid
 Maintain the temperature as it was at the
start of process
 Add some quantity of water to make original
volume of test liquid
 Resulted distillate may be clear or not if not
then clear it by adding the talc or CaCO3
 Now determine the specific gravity at 25 oC
 So we can calculate the %age v/v of alcohol.
b) For liquids presumed to contain 50% of alcohol
or less:
Adnan Sarwar Chaudhary
 By using pipette transfer liquid dosage form
in distilling apparatus equal to 25ml and
note
 the temperature
 Add double volume of water as compared to
sample
 Collect a volume of distillate 2ml less than,
twice the volume of original test liquid
 Maintain temperature of distillate as of
original temperature
 Add some amount of water to double
exactly the volume as taken at start
 Mix and note specific gravity
 Now determine the %age of alcohol v/v.
 That must be ½ of original liquid examine
c) For liquids presumed to contain more than 50% of
alcohol:
Adjust the specimen under examination by diluting
it with water up to 25% ad then performed process
as above.
II. Gas Liquid Chromatography Method:
USP Reference Standards
USP Alcohol Determination—Acetonitrile RS
USP Alcohol Determination—Alcohol RS
Method IIa
Apparatus
Gas chromatography specification
1. Flame-ionization detector
2. 4-mm × 1.8-m glass column packed with
100- to 120-mesh chromatographic column
packing support S3
3. Nitrogen or helium as the carrier.
Temperature specification
1. Prior to use, condition the column overnight
at 235oC with a slow flow of carrier gas.
2. The column temperature is maintained at
120o
3. The injection port and detector temperatures
are maintained at 210o.
4. Adjust the carrier flow and temperature so
that acetonitrile, the internal standard, elutes
in 5 to 10 minutes.
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Solutions:  Six replicate injections of the Standard

Preparation show a relative standard
 Test Stock Preparation: Dilute the specimen
deviation of not more than 2.0% in the ratio
under examination stepwise with water to
of the peak of alcohol to the peak of the
obtain a solution containing approximately 2%
internal standard.
(v/v) of alcohol.
 Test Preparation: Pipet 5 mL each of the Test
Method IIb
Stock Preparation and the USP Alcohol
Determination— Apparatus:
 Acetonitrile RS [Alternatively, a 2% aqueous
solution of acetonitrile of suitable quality may Gas chromatography specification
1. Split injection port with a split ratio of 5:1
be used as the internal standard solution] into a
2. A flame-ionization detector
50-mL volumetric flask, dilute with water to
3. 0.53-mm × 30-m capillary column coated with a
volume, and mix. 3.0-µm film of phase G43.
 Standard Preparation: Pipet 5 mL each of the 4. Helium is used as the carrier gas at a linear
USP Alcohol Determination—Alcohol RS and velocity of 34.0 cm per second.
the USP
 Alcohol Determination—Acetonitrile RS Temperature specification
1. The chromatograph is programmed to maintain
[Alternatively, a 2% aqueous solution of
the column temperature at 50o for 5 minutes
acetonitrile of suitable quality may be used as
2. Then to increase the temperature at a rate of
the internal standard solution] into a 50-mL
10o per minute to 200o, and maintain at this
volumetric flask, dilute with water to volume,
temperature for 4 minutes.
and mix.
3. The injection port temperature is maintained
Procedure at 210o
4. The detector temperature at 280o.
 Inject about 5 µL each of the Test Preparation
and the Standard Preparation, in duplicate, into Solutions:
the gas chromatograph, record the
 Test Stock Preparation: Dilute the specimen
chromatograms, and determine the peak
under examination stepwise with water to
response ratios.
obtain a solution containing approximately
 Calculate the percentage of alcohol (v/v) in the
2% (v/v) of alcohol.
specimen under test according to the formula:
 Test Preparation: Pipet 5 mL each of the
CD(RU / RS)
Test Stock
 C is the labeled concentration of USP
 Preparation and the USP Alcohol
Alcohol Determination—Alcohol RS
Determination— Acetonitrile RS
 D is the dilution factor (the ratio of the
[Alternatively, a 2% aqueous solution of
volume of the Test Stock Preparation to the
acetonitrile of suitable quality may be used
volume of the specimen taken);
as the internal standard solution] into a 25-
 RU and RS are the peak response ratios
mL volumetric flask, dilute with water to
obtained from the Test Preparation and the
volume, and mix.
 Standard Preparation, respectively.
 Standard Preparation: Pipet 5 mL each of
the USP Alcohol Determination—Alcohol
System Suitability Test
RS and the USP
 In a suitable chromatogram, the resolution  Alcohol Determination—Acetonitrile RS
factor, R, is not less than 2; [Alternatively, a 2% aqueous solution of
 The tailing factor of the alcohol peak is not acetonitrile of suitable quality may be used
greater than 2.0
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as the internal standard solution] into a 25-
mL volumetric flask, dilute with water to
volume, and mix.
Procedure
 Inject about 0.2 to 0.5 µL each of the Test
Preparation and the Standard Preparation, in
duplicate, into the gas chromatograph,
record the chromatograms, and determine
the peak response ratios.
 Calculate the percentage of alcohol (v/v) in
the specimen under test according to the
formula:
CD(RU / RS)
o C is the labeled concentration of USP
Alcohol Determination—Alcohol RS
o D is the dilution factor (the ratio of the
volume of the Test Stock Preparation to the
volume of the specimen taken);
o RU and RS are the peak response ratios
obtained from the Test Preparation and the
Standard Preparation, respectively.
System Suitability Test:
 In a suitable chromatogram, the resolution
factor, R, between alcohol and the internal
standard is not less than 4
 The tailing factor of the alcohol peak is not
greater than 2.0
 Six replicate injections of the Standard
Preparation show a relative standard
deviation of not more than 4.0% in the ratio
of the peak of alcohol to the peak of the
internal standard.Used only when specified
in individual monograph.
3. FOR EMULSION
i. Type of Emulsion:
Type of emulsion is determined whether it w/o or
o/w as follow.
a) Physical examination test:
Emulsion is applied on skin and if is easily
removable then it is o/w emulsion and if it is not
easily removable then it is w/o emulsion.
Adnan Sarwar Chaudhary
b) Dilution test:
Suitable diluents like water are added and stability
is checked if it is stable then it is o/w type of
emulsion if emulsion break down then it is w/o
emulsion.
c) Conductivity test:
This test is based on conduction of electric current
in emulsion for which a pair of electrodes is applied
in emulsion and outer circuit is completed. If
electricity passed then it is o/w type of emulsion
and if not pass then it is w/o type of emulsion.
d) Dye test:
Dye is added to emulsion and it may be water
soluble (methylene blue) will distribute throughout
in o/w emulsion or may be Oil soluble (scarlet red)
will distribute throughout in w/o emulsion.
e) CoCl2 test:
Simply dip the filter paper into solution of CoCl2, it
will turn blue upon drying and dip it into emulsion
for test. If paper turns pink emulsion is w/o and if it
remains unchanged then it is o/w type of emulsion.
f) Fluorescence test:
As oils are exposed to UV rays, they become
fluorescent so when o/w emulsion exposed to
UVlight emulsion globules become fluorescent and
when w/o type of emulsion exposed to UV light the
continuous phase will be fluorescent.
ii. Dispersibility/pourability:
It is used to check the spreadability & pourability of
emulsion.
Process:
If continuous phase is oil soluble, emulsion is
diluted with oil & if continuous phase is water
soluble then emulsion is diluted with water.
And effects of dispersibility on skin are checked by
spreading the emulsion on skin.
iii. Physical stability of emulsion:
It is checked by visual inspection of emulsion in
which coalescence of internal phase, and creaming,
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elegance of emulsion, odour, colour and other
physical properties are checked.
iv. Water content:
Water content of emulsion is determined by titration
method called Karl Fischer titration
It is a classic titration method that uses volumetric
titration to determine trace amounts of water in a
sample.
v. Globule size determination:
Use for assessment of shelf life of emulsion in
which we consider about the
a. Uniform size of globule
b. Uniform distribution of globule
Globule size analysis is carried out by
a. Microscopic measurement:
It gives an average value dependent on the no of
globules of each size
b. Coulter counter:
It measures the globule volume and check the
uniformity of distribution.
4. FOR SUSPENSION
i. Particle size determination of
suspension:
mixed with the equal quantity of glycerol and
further dilution is carried out by mixture of glycerol
and water.
unt the sample material on slide and
examine under microscope, measure the diameter of
particles above the maximum size permitted in
individual monograph.
1000 particles other diluents may be paraffin.
Adnan Sarwar Chaudhary
ii. Sedimentation volume of
suspensions:
Physical stability of suspension depends upon
a. Size of particle
b. Suspending agent (↑ Suspending
agent, ↑ Viscosity, ↑ Sedimentation
volume, ↑ Stability)
Following two parameter determine the
pharmaceutical acceptability of suspensions,
a. Sedimentation volume
b. Degree of flocculation
Sedimentation volume F can be defined as ―The
ratio of final or ultimate volume of sediment Vu to
the original volume of suspension Vo before
settling‖.
F=
 Sedimentation volume can have values
ranging from less than 1 to greater than 1.
 When volume of sediment in a flocculated
suspension equals the original volume of
suspension then F=1.
 Such a product is said to be in ―flocculation
equilibrium‖ and shows no clear supernatant
on standing it is pharmaceutically
acceptable.
iii. Crystal growth in suspension:
It is a common cause of deterioration of suspension.
Process:
 Crystal growth is achieved by simulating the
temperature fluctuation under normal
storage conditions.
 But at greatly increased frequency as daily
variation of temperature has been
reproduced by cycling time of 16 min.
 Maintain temperature at 23-33 oC, alter
temperature from this range after every 16
minutes then crystal growth
 Crystal growth depends upon the particles
concentration, the bulk particles solubility,
the slope of solubility curve, the temperature
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fluctuation range and frequency of calculate density by using the constants A and B
fluctuation. described below; and
 a means to measure and/or control the
Weight per ml temperature of the oscillating transducer
containing the liquid to be tested.
The weight per milliliter of a liquid is the weight, in
gm., of 1 ml of a liquid when weighed in air at The oscillation period is a function of the spring
25°C, unless otherwise specified. constant (c) and the mass of the system:

T 2 = {(M/c) + [( r × V)/c]} × 4p 2 where r is the

METHOD I
density of the liquid to be tested, M is the mass of
Procedure the tube, and V is the volume of the filled tube.
Select a scrupulously clean, dry pycnometer that
Introduction of two constants A = c/(4p 2 × V) and
previously has been calibrated by determining its
B = (M/V) leads to the classical equation for the
weight and the weight of recently boiled water
oscillating transducer:
contained in it at 25°. Adjust the temperature of the
liquid to about 20°, and fill the pycnometer with it. r=A×T2–B

Adjust the temperature of the filled pycnometer to The specific gravity of the liquid is given by the
25°, remove any excess liquid, and weigh. When formula:
the monograph specifies a temperature different
from 25°, filled pycnometers must be brought to the r (L) / r (W)
temperature of the balance before they are weighed. where r (L) and r (W) are the densities of the liquid
Subtract the tare weight of the pycnometer from the and water, respectively, both determined at 25°,
filled weight. unless otherwise directed in the individual
The specific gravity of the liquid is the quotient monograph.
obtained by dividing the weight of the liquid
contained in the pycnometer by the weight of water
contained in it, both determined at 25°, unless
otherwise directed in the individual monograph.

METHOD II
The procedure includes the use of the oscillating
transducer density meter. The apparatus consists of
the following:

 a U-shaped tube, usually of borosilicate glass,

which contains the liquid to be examined;
 a magneto-electrical or piezo-electrical
excitation system that causes the tube to
oscillate as a cantilever oscillator at a
characteristic frequency depending on the
density of the liquid to be examined;
 a means of measuring the oscillation period (T),
which may be converted by the apparatus to
give a direct reading of density or used to

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QUALITY CONTROL
OF SUPPOSITORIES

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Physical Testing Now calculate the percentage deviation by

1. Visual Examination following formula.
2. Weight Variation Test
3. Melting Range Test
4. Breaking Test
5. Disintegration Test (Liquefaction Test or Acceptance criteria
Softening Test) Not more than 2 of the individual masses deviate
from the average mass by more than 5% and no
Chemical Testing
suppository should deviate from average weight by
1. Content Uniformity Test
more than 10%. All weight must be range in 10%.
2. Dissolution Test

Melting Range Test

Physical Testing
This test is also called macro melting range test.
This test is a measure of time it takes for entire
Visual Examination: suppository to melt when immersed in a constant
Visual evaluation of suppositories is necessary and temperature water bath, i.e. 37 oC.
important to check for the absence of fissuring,
Purpose:
pitting, fat blooming, exudation, sedimentation and
This test is performed to check either suppository is
migration of active ingredient.
melt within body cavity at optimum time or not.
1. Purpose
Apparatus:
Purpose of this test is to check uniformity between
USP-Tablet Disintegration Apparatus is used to find
batches and physical quality assessment.
melting range.
2. Shape & Size
Procedure:
Suppositories must be of proper shape and size
Set the apparatus and maintain the temperature at 37
because change in shape affects the suppositories. o
C. Immerse the suppository in glass tube and
Change in shape can effect the formulation
immerse the tube in water bath and run the
Check the shape of the suppositories to see whether apparatus.
it is consistent or intact or not
Time for the entire suppository to melt or dispersed
3. Color or disintegrated in surrounding water is measured.
Intensity, nature and homogeneity of color should
Disintegration:
be checked and verified.
Suppository is considered to be disintegrated when:
4. Odor
 It is completely dissolved or disintegrated.
Must be of desired odour because change in odour
 It is dispersed into its components or parts.
indicate the degradation process.
 It became soft in shape with the formation of
5. Stability core which is not resistant to pressure of
Check the stability there should be no crack and glass rod.
bubbles in the suppositories.
Acceptance criteria:
Fat base: 30 minutes
Weight Variation Test
Water base: 60 minutes.
Weigh 20 suppositories individually and calculate
average weight.

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Breaking Test
Purpose:
This test applied to suppositories and passeries
based on fatty excipients. It is not suited to
suppositories and passeries based on hydrophilic
excipients such as gelatin-glycerol mixture.
This test is design to check:
 Hardness
 Fragility
 Brittlness
Apparatus:
 A thermostattted chamber closed in front by
a glass window and containing a device that
is to hold the suppository or passery.
 Two opposite jaws, the upper jaw
descending vertically toward the lower jaw.
The crushing surfaces of the jaw are flat
perpendicular, to the direction of movement
and larger than zone of contact with the
suppository or passery.
 A plastic sample holder is fixed in the center
of the jaw (half a holder in each jaw).
 The upper jaw (top pressure block) is
connected to suspension to which can be
added each of which weight 200g. The
initial mass of the device is 600g. Crushing
of the sample is carried out by successive
adding 200g disc to the initial mass of 600g.
Method:
 Check that apparatus is vertical. Heat the
thermostattted chamber to 25 oC.
 The dosage form to be tested has been
maintained for at least 24 hours at the
required measuring temperature. Place the
suppository or passery vertically between
the jaws in the sample holder with the point
upwards.
 The top pressure block of the suspension
loading rod is carefully positioned and the
test chamber is closed with its glass
window; for each determination position the
Adnan Sarwar Chaudhary
suppository or passery in the same manner
with respect to force applied.
 Wait for 1min and add the first 200g disc,
again wait for 1min and add another disc.
Repeat the operation until the suppository or
passery collapses.
 The mass required to crush the suppository
or pessary is calculated by the sum of the
masses weighing on the suppository or
pessary when it collapses (including the
initial mass of the device) assessed as
following
o If the suppository or passery collapses
within 20 seconds of placing the last disc
do not take its mass into account.
o If the suppository or passery collapse
between 20 seconds and 40 seconds of
placing the last disc, use only half of
this mass in the calculations i.e. 100g.
o If the suppository or passery remains
uncrushed for more than 40 sec after the
last disc is placed, use all the mass in
the calculation.
 Carry out each measurement on 10
suppositories or 10 passeries, making sure
that no residue remains before each
determination.
Disintegration Test
(Liquefaction Test or
Sogtening Test)
Suppository is considered to be disintegrated when:
 It is completely dissolved or disintegrated.
 It is dispersed into its components or parts.
 It became soft in shape with the formation of
core which is not resistant to pressure of
glass rod.
Methods:
There are two methods are used for disintegration:
1. Method-I
2. Method-II
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Method-I (water & fat base) Test criteria:

The test is carried out using three such apparatuses
This method is used for both water and fat base
each containing a single sample.
suppositories.
Procedure:
Apparatus:
 Each apparatus is placed in a beaker with a
The apparatus consists of 2 main parts:
capacity of at least 4 L filled with water
1. Cylinder/vessel/beaker of glass or suitable maintained at 36-37 °C, unless otherwise
transparent plastic, specified in monograph.
2. Metal device with two perforated stainless  The beaker is fitted with a slow stirrer and a
metal discs which contains 39 holes or device that will hold the cylinders vertically
perforations not less than 90 mm below the surface of the
water and allow them to be inverted without
.
emerging from the water.
 Place one suppository on the lower disc of a
device.
 Invert the apparatus every 10 min. and note
the time of disintegration.

Acceptance criteria:
 Fat base: 30 minutes
 Water base: 60 minutes.

Method-II (fat base)

This test is used for the disintegration of fat base or
lipophilic suppositories.

Priniciple:
This test is intended to determine the time
(under defined conditions) which elapses until a
suppository, maintained in a water, soften to extent
so that no longer offers resistance when a defined
weight is applied.

Apparatus
Two types of apparatus are used:

1. Apparatus-A
Assembly: 2. Apparatus-B
 Metal device is inserted into cylinder and
attach to the rim of the cylinder with 3
Apparatus-A:
spring clips. It consists of two parts:
1. Glass tube
 Each disc containing 39 holes 4 mm in
2. Glas rod
diameter held 30mm a part evenly spaced in
a concentric pattern. Glass tube:
1. It is tube of glass having internal diameter
15.5mm.
2. Flat bottom.
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3. Length is 140mm.
4. Tube is closed with removeable plastic
cover having opening of 5.2 mm for rod.
Glass rod
Upper part is made of plastic or metal having
diameter 5.0mm with a weight disc. Upper part
containing sliding mark ring when the rod is
introduce into glass tube it touches the bottom,
sliding mark ring is adjusted to concide with upper
level of the plastic cover.
Lower part made up of plastic and wider having
diameter12mm.
Metal needle is fixed at flat lower side with 2mm
length and 1mm diameter.
Procedure:
 Place glass tube having 10ml of water in
water bath maintained at 36.5±0.5 oC.
 Fix the glass tube vertically and immersed
at a depth of at least 7cm below the surface
but without touching the bottom of water
bath.
 Introduce a suppository tip downward
followed by the rod with a free gliding
plastic cover into the glass tube until the
metal needle touches the flat end of the
suppository.
 Put the cover on the tube and start the time
measurement.
Adnan Sarwar Chaudhary
 Note the time which passes until the rod
sinks downward to the bottom of glass
tube and sliding mark reach the upper level
of the plastic cover.
Apparatus-B:
It consists of three parts:
 A water bath
 An inner tube which is inserted in water bath
and fixed with stopper. The inner tube is
closed by a stopper at the bottom.
 Thermometer: The apparatus is also fixed
with a thermometer to note the temperature.
 Two insets are available
1. Glass rod (C1): It is in the form of a tube
sealed at both ends, carrying a ring at its
lower end weighted with lead shot, which
has weight of 30±0.4g.
2. A penetration inset (C2): it consists of a
rod (7.5±0.1g) in a tube which has an
enlargement for the suppository both made
of stainless steel.
Method:
 Pour 5ml of water at 36.5±0.5ᵒC into the
inner tube (A).
 Introduce a suppository with the tip
downward.
 Place the inset at the suppository.
 Note the time which elapses between this
moment and the moment when the lower rim
end of the glass rod (C1) or penetration inset
rod (C2) reaches the narrow part of the
inner glass tube. Melting or dissolution is
then considered as complete.
Acceptance and rejection criteria:
 Fat base: 30 minutes
 Water base: 60 minutes.
[Difference between apparatus A and apparatus
B is that apparatus B also has a penetration
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inset]
Chemical Tests
Content Uniformity (EP)
This test is used to confirm that every unit should
contain same amount of drug or active ingredients
with little variation in the batch.
Principle:
The test of the uniformity of the content of single
dose preparations is based on the assay of
individual contents of active substance(s) of a
number of dosage units to determine whether the
individual contents are within the limits set with
reference to the average content of the sample.
Method:
 Using a suitable analytical method,
determine the individual contents of active
substance(s) of 10 dosage units taken at
random.
 Now apply the criteria of Test A, Test B or
Test C as specified in the monograph for
any specified dosage form.
 Use Test B for Suppositories given below:
Test B
 Take 10 tablets randomly and determine the
individual content of active substance. The
preparation compiles with the test if not
more than 1 individual content is outside the
limit of ±15% of the average content and
none is the outside the limit of ±25% of the
average content.
Adnan Sarwar Chaudhary
 The preparation fail to comply with the test
if more than 3 individual contents are
outside the limit of
 ±15% of the average content or if one or
more individual contents are outside the
limit of 75% of the average content.
 If 2 or more individual contents are outside
the limits of ±15% but within the limit of
±25%. Then determine the individual
contents of 20 dosage units taken at random.
 The preparation compiles with the test if not
more than 3 individual contents of 30 units
are outside the limit of the average content.
Dissolution Test:
The test determines the amount of dosage form that
gets dissolve in body fluid per unit time or it is the
measure of rate of drug release from the
suppositories.
Type of apparatus:
Two types of apparatus are used
i. Suppository dialysis cell
ii. Stationary basket
Suppository dialysis cell:
It is also called flow through cell apparatus.
Lipophilic suppositories are tested in it.
Stationary basket:
This apparatus is used for hydrophilic suppositories.
This apparatus consists of a basket and rotating
paddles.
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QUALITY CONTROL
OF STERILE
PRODUCTS
(PARENTERALS)

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implantation into the human or animal body.

STERILE PRODUCTS Parenteral preparations are supplied in glass, plastic
Sterile products are the dosage form of therapeutic containers and prefilled syringes with closure are
agents that are free from viable micro-organisms. made up of plastic or elastomers

These sterile products include the followings: QUALITY CONTROL TESTS

1. Parenterals Six types of test are used to check the quality if sterile
2. Ophthalmic products which are as follow:
3. Irrigating preparations 1. Pyrogen Test
2. Clarity Test
Of these parenteral products are unique among the
3. Sterility Test
dosage forms of the drugs because they are injected 4. Leaker‘s Test
through skin or mucous membranes into the internal 5. Safety Test
body compartments. 6. Content Uniformity Test.

Ophthalmics (BP):
These are the sterile liquids, semisolids or solid Sterility Test
preparations intended for administration upon the
The test for sterility is carried out under
Eyeball and/ or to the conjunctiva or for insertion in
aseptic conditions. In order to achieve such
the conjunctival sac.
conditions, the test environment has to be adapted
Categories of ophthalmics: to the way in which the sterility test is performed.
The precautions taken to avoid contamination are
1. Eye drops such that they do not affect any micro-organisms
2. Eye lotions which are to be revealed in the test.
3. Powders for eye drops The working conditions in which the tests are
4. Powders for eye lotions performed are monitored regularly by appropriate
5. Semisolid eye preparations sampling of the working area and by carrying out
6. Ophthalmic inserts appropriate controls.
Irrigating Preparations: Two methods are used for sterilization of
These solutions are applied topically to bathe open parenterals:
wounds and body cavities. They are sterile solutions
for single use only. 1. Membrane Filteration
2. Direct Inoculation
 Water for irrigation is sterilized distilled
water that is free of pyrogens. The procedures are applicable for
 The water is packed in containers and is determining whether a Pharmacopeial article
intended for use on one occasion only. purporting to be sterile complies with the
 The containers are sealed and sterilized by requirements set forth in the individual monograph
moist heat. with respect to the test for sterility. Pharmacopeial
articles are to be tested by the Membrane Filtration
Examples: method under Test for Sterility of the Product to be
1. 0.9% w/v sodium chloride solution examined where the nature of the product permits.
2. Sterile water for irrigation. If the membrane filtration technique is unsuitable,
use the Direct Inoculation of the Culture Medium
PARENTERAL PREPARATIONS (BP)
method under Test for Sterility of the Product to be
Parenteral preparations are sterile preparations
examined. All devices, with the exception of
intended for administration by injection, infusion or
Devices with Pathways Labeled Sterile, are tested
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using the Direct Inoculation of the Culture Medium
method. Provisions for retesting are included under
Observation and Interpretation of Results.
Because sterility testing is a very exacting
procedure, where asepsis of the procedure must be
ensured for a correct interpretation of results, it is
important that personnel be properly trained and
qualified. The test for sterility is carried out under
aseptic conditions. In order to achieve such
conditions, the test environment has to be adapted
to the way in which the sterility test is performed.
The precautions taken to avoid contamination are
such that they do not affect any microorganisms that
are to be revealed in the test. The working
conditions in which the tests are performed are
monitored regularly by appropriate sampling of the
working area and by carrying out appropriate
controls.
These Pharmacopeial procedures are not by
themselves designed to ensure that a batch of
product is sterile or has been sterilized. This is
accomplished primarily by validation of the
sterilization process or of the aseptic processing
procedures.
When evidence of microbial contamination
in the article is obtained by the appropriate
Pharmacopeial method, the result so obtained is
conclusive evidence of failure of the article to meet
the requirements of the test for sterility, even if a
different result is obtained by an alternative
procedure.
MEDIA
Prepare media for the tests as described
below or dehydrated formulations may be used
provided that, when reconstituted as directed by the
manufacturer or distributor, they meet the
requirements of the Growth Promotion Test of
Aerobes, Anaerobes, and Fungi. Media are
sterilized using a validated process.
The following culture media have been
found to be suitable for the test for sterility. Fluid
Thioglycollate Medium is primarily intended for the
culture of anaerobic bacteria. However, it will
Adnan Sarwar Chaudhary
also detect aerobic bacteria. Soybean–Casein Digest
Medium is suitable for the culture of both
fungi and aerobic bacteria.
Fluid Thioglycollate Medium
L-Cystine 0.5 g
Sodium Chloride 2.5 g
Dextrose (C6H12O6·H2O) 5.5/5.0 g
Agar, granulated (moisture content 0.75 g
not exceeding 15%)
Yeast Extract (water-soluble) 5.0 g
Pancreatic Digest of Casein 15.0 g
Sodium Thioglycollate 0.5 g
or Thioglycolic Acid 0.3 mL
Resazurin Sodium Solution (1 in 1.0 mL
1000), freshly prepared
Purified Water 1000 Ml
Mix the L-cysteine, sodium chloride,
dextrose, yeast extract, and pancreatic digest of
casein with the purified water, and heat until
solution is affected. Dissolve the sodium
thioglycollate or thioglycolic acid in the solution
and, if necessary, add 1 N sodium hydroxide so that,
after sterilization, the solution will have a pH of 7.1
± 0.2. If filtration is necessary, heat the solution
again without boiling, and filter while hot through
moistened filter paper.
Add the resazurin sodium solution, mix, and
place the medium in suitable vessels that provide a
ratio of surface to depth of medium such that
not more than the upper half of the medium has
undergone a color change indicative of oxygen
uptake at the end of the incubation period. Sterilize
using a validated process. If the medium is
stored, store at a temperature between 2 and 25° in a
sterile, airtight container. If more than the
upper one-third of the medium has acquired a pink
color, the medium may be restored once by
heating the containers in a water-bath or in free-
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flowing steam until the pink color disappears and
by cooling quickly, taking care to prevent the
introduction of nonsterile air into the container.
Fluid Thioglycollate Medium is to be incubated at
32.5 ± 2.5°.
Alternative Thioglycollate Medium
Prepare a mixture having the same
composition as that of the Fluid Thioglycollate
Medium, but omitting the agar and the resazurin
sodium solution, sterilize as directed above, and
allow cooling prior to use. The pH after sterilization
is 7.1 ± 0.2. Incubate under anaerobic conditions for
the duration of the incubation period. Alternative
Fluid Thioglycollate Medium is to be incubated at
32.5 ± 2.5°.
Soybean–Casein Digest Medium
Pancreatic Digest of Casein 17.0 g
Papaic Digest of Soybean Meal 3.0 g
Sodium Chloride 5.0 g
Dibasic Potassium Phosphate 2.5 g
Dextrose (C6H12O6·H2O) 2.5/2.3 g
Purified Water 1000 mL
Dissolve the solids in the Purified Water, heating
slightly to effect a solution. Cool the solution to
room temperature, and adjust the pH with 1 N
sodium hydroxide so that, after sterilization, it will
have a pH of 7.3 ± 0.2. Filter, if necessary to clarify,
dispense into suitable containers, and sterilize
using a validated procedure. Store at a temperature
between 2° and 25° in a sterile well-closed
container, unless it is intended for immediate use.
Soybean–Casein Digest Medium is to be incubated
at 22.5 ± 2.5°.
Suitability Tests
The media used comply with the following tests,
carried out before, or in parallel, with the test on
the product to be examined.
Adnan Sarwar Chaudhary
STERILITY
Confirm the sterility of each sterilized batch of
medium by incubating a portion of the media at the
specified incubation temperature for 14 days. No
growth of microorganisms occurs.
GROWTH PROMOTION TEST
OF AEROBES, ANAEROBES,
AND FUNGI
Test each lot of ready-prepared medium and each
batch of medium prepared either from dehydrated
medium or from ingredients. Inoculate portions of
Fluid Thioglycollate Medium with a small number
(not more than 100 cfu) of the following
microorganisms, using a separate portion of
medium for each of the following species of
microorganism: Clostridium sporogenes,
Pseudomonas aeruginosa, and Staphylococcus
aureus. Inoculate portions of Alternative Fluid
Thioglycollate Medium with a small number (not
more than 100 cfu) of Clostridium sporogenes.
Inoculate portions of Soybean–Casein Digest
Medium with a small number (not more than 100
cfu) of the following microorganisms, using a
separate portion of medium for each of the
following species of microorganism: Aspergillus
niger, Bacillus subtilis, and Candida albicans.
Incubate for not more than 3 days in the case of
bacteria and not more than 5 days in the case of
fungi. The media are suitable if a clearly visible
growth of the microorganisms occurs.
STORAGE
If prepared media are stored in unsealed containers,
they can be used for 1 month, provided that they are
tested for growth promotion within 2 weeks of the
time of use and that color indicator requirements are
met. If stored in tight containers, the media can be
used for 1 year, provided that they are tested for
growth promotion within 3 months of the time of
use and that the color indicator requirements are
met.
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DILUTING AND RINSING
FLUIDS FOR MEMBRANE
FILTRATION
Fluid A
PREPARATION
Dissolve 1 g of peptic digest of animal tissue in
water to make 1 L, filter or centrifuge to clarify, if
necessary, and adjust to a pH of 7.1 ± 0.2. Dispense
into containers, and sterilize using a validated
process.
PREPARATION FOR PENICILLINS OR
CEPHALOSPORINS
Aseptically add to the above Preparation, if
necessary, a quantity of sterile -lactamase sufficient
to inactivate any residual antibiotic activity on the
membranes after the solution of the test specimen
has been filtered (see Media for Penicillins or
Cephalosporins).
Fluid D
To each L of Fluid A add 1 mL of polysorbate 80,
adjust to a pH of 7.1 ± 0.2, dispense into containers,
and sterilize using a validated process. Use this
fluid for articles containing lecithin or oil, or for
devices labeled as ―sterile pathway.‖
Fluid K
Dissolve 5.0 g of peptic digest of animal tissue, 3.0
g of beef extract, and 10.0 g of polysorbate 80 in
water to make 1 L. Adjust the pH to obtain, after
sterilization, a pH of 6.9 ± 0.2. Dispense into
containers, and sterilize using a validated process.
VALIDATION TEST
Carry out a test as described below under Test for
Sterility of the Product to be Examined using
exactly the same methods, except for the following
modifications.
Membrane Filtration
After transferring the content of the container or
containers to be tested to the membrane, add an
inoculum of a small number of viable
microorganisms (not more than 100 cfu) to the final
portion of sterile diluent used to rinse the filter.
Adnan Sarwar Chaudhary
Direct Inoculation
After transferring the contents of the container or
containers to be tested (for catgut and other surgical
sutures for veterinary use: strands) to the culture
medium, add an inoculum of a small number of
viable microorganisms (not more than 100 cfu) to
the medium.
In both cases use the same microorganisms as those
described above under Growth Promotion Test of
Aerobes, Anaerobes, and Fungi. Perform a growth
promotion test as a positive control.
Incubate all the containers containing medium for
not more than 5 days. If clearly visible growth of
microorganisms is obtained after the incubation,
visually comparable to that in the control vessel
without product, either the product possesses no
antimicrobial activity under the conditions of the
test or such activity has been satisfactorily
eliminated. The test for sterility may then be carried
out without further modification.
If clearly visible growth is not obtained in the
presence of the product to be tested, visually
comparable to that in the control vessels without
product, the product possesses antimicrobial activity
that has not been satisfactorily eliminated under the
conditions of the test. Modify the conditions in
order to eliminate the antimicrobial activity, and
repeat the validation test.
This validation is performed (a) when the test for
sterility has to be carried out on a new product; and
(b) whenever there is a change in the experimental
conditions of the test. The validation may be
performed simultaneously with the Test for Sterility
of the Product to be Examined.
TEST FOR STERILITY OF THE
PRODUCT TO BE EXAMINED
Number of Articles to Be Tested
Unless otherwise specified elsewhere in this chapter
or in the individual monograph, test the number of
articles specified in Table 3. If the contents of each
article are of sufficient quantity (see Table 2), they
may be divided so that equal appropriate portions
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are added to each of the specified media. [NOTE than 300 mg not
Perform sterility testing employing two or more of less than 50 mg
the specified media.] If each article does not contain
sufficient quantities for each medium, use twice the 300 mg–5 g 150 mg
number of articles indicated in Table 3. Greater than 5 g 500 mg
Table 2. Minimum Quantity to be Used for Each Devices
Medium
Minimum Quantity to Catgut and other surgical
3 sections of a strand
be Used sutures for
(each 30-cm long)
Quantity per Container (unless otherwise veterinary use
justified and
authorized) Surgical
dressing/cotton/gauze (in 100 mg per package
Liquids (other than anitbiotics) packages)

The whole contents of Sutures and other

Less than 1 mL
each container individually packaged The whole device
single-use material
Half the contents of
each container, but The whole device, cut
1–40 mL
not Other medical devices into pieces or
less than 1 mL disassembled

Greater than 40 mL, and

not greater 20 mL Table 3. Minimum Number of Articles to be Tested
than 100 mL in Relation to the Number of Articles in the Batch
Minimum Number of
10% of the contents Items to be Tested for
Number of Items in the
Greater than 100 mL of the container, but Each Medium (unless
Batch
not less than 20 mL otherwise justified and
authorized) *

Antibiotic liquids 1 mL
Parenteral preparations
The whole contents of
Other preparations soluble
each container to
in water or in
provide not less than
isopropyl myristate 10% or 4
200 mg
containers,
Not more than 100 containers
Insoluble preparations, Use the contents of whichever is the
creams, and each container to greater
ointments to be provide not less than
More than 100 but not more
suspended or emulsified 200 mg 10 containers
than 500 containers
Solids
2% or 20
The whole contents of More than 500 containers containers,
Less than 50 mg
each container whichever is less

Half the contents of For large-volume parenterals 2% or 10

50 mg or more, but less
each container, but containers,
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whichever is less
Antibiotic solids
Pharmacy bulk packages (<5
20 containers
g)
Pharmacy bulk packages ( 5
6 containers
g)
See Bulk solid
Bulks and blends
products
Ophthalmic and other noninjectable preparations
5% or 2 Culture Medium with the product to be examined.
containers,
Not more than 200 containers
whichever is the
greater
More than 200 containers 10 containers
If the product is presented in the form of single-
dose containers, apply the scheme shown above for
preparations for parenteral use.
Devices
2% or 5 packages,
Catgut and other surgical whichever is the
sutures for greater, up to a
veterinary use maximum total of
20 packages
10% or 4 articles,
Not more than 100 articles whichever is
greater
More than 100, but not more
10 articles
than 500 articles
2% or 20 articles,
More than 500 articles
whichever is less
Bulk solid products
Up to 4 containers Each container
More than 4 containers, but 20% or 4 suitable for carrying out the incubation after adding
not more than 50 containers containers,
whichever is
Adnan Sarwar Chaudhary
greater
2% or 10
containers,
More than 50 containers
whichever is
greater
If the contents of one container are enough to
*
inoculate the two media, this column gives the
number of containers needed for both the media
together.
The test may be carried out using the technique of
Membrane Filtration or by Direct Inoculation of the
Appropriate negative controls are included. The
technique of membrane filtration is used whenever
the nature of the product permits; that is, for
filterable aqueous preparations, for alcoholic or oily
preparations, and for preparations miscible with, or
soluble in, aqueous or oily solvents, provided these
solvents do not have an antimicrobial effect in the
conditions of the test.
Membrane Filtration
Use membrane filters having a nominal pore size
not greater than 0.45 µm whose effectiveness to
retain microorganisms has been established.
Cellulose nitrate filters, for example, are used for
aqueous, oily, and weakly alcoholic solutions; and
cellulose acetate filters, for example, are used for
strongly alcoholic solutions. Specially adapted
filters may be needed for certain products (e.g., for
antibiotics).
The technique described below assumes that
membranes about 50 mm in diameter will be used.
If filters of a different diameter are used, the
volumes of the dilutions and the washings should be
adjusted accordingly. The filtration apparatus and
membrane are sterilized by appropriate means. The
apparatus is designed so that the solution to be
examined can be introduced and filtered under
aseptic conditions: it permits the aseptic removal of
the membrane for transfer to the medium, or it is
the medium to the apparatus itself.
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AQUEOUS SOLUTIONS
If appropriate, transfer a small quantity of a
suitable, sterile diluent such as Fluid A (see Diluting
and Rinsing Fluids for Membrane Filtration) onto
the membrane in the apparatus and filter. The
diluent may contain suitable neutralizing substances
and/or appropriate inactivating substances, for
example, in the case of antibiotics.
Transfer the contents of the container or containers
to be tested to the membrane or membranes, if
necessary, after diluting to the volume used in the
Validation Test with the chosen sterile diluent, but
using not less than the quantities of the product to
be examined prescribed in Tables 2 and 3. Filter
immediately. If the product has antimicrobial
properties, wash the membrane not less than three
times by filtering through it each time the volume of
the chosen sterile diluent used in the Validation
Test. Do not exceed a washing cycle of 5 times 200
mL, even if during validation it has been
demonstrated that such a cycle does not fully
eliminate the antimicrobial activity. Transfer the
whole membrane to the culture medium or cut it
aseptically into two equal parts, and transfer one
half to each of two suitable media. Use the same
volume of each medium as in the Validation Test.
Alternatively, transfer the medium onto the
membrane in the apparatus. Incubate the media for
not less than 14 days.
SOLUBLE SOLIDS (OTHER THAN
ANTIBIOTICS)
Use for each medium not less than the quantity
prescribed in Tables 2 and 3 of the product
dissolved in a suitable solvent, such as Fluid A
(Diluting and Rinsing Fluids for Membrane
Filtration), and proceed with the test as described
above for Aqueous Solutions using a
membrane appropriate to the chosen solvent.
OILS AND OILY SOLUTIONS
Use for each medium not less than the quantity of
the product prescribed in Tables 2 and 3. Oils and
oily solutions of sufficiently low viscosity may be
filtered without dilution through a dry membrane.
Viscous oils may be diluted as necessary with a
suitable sterile diluent such as isopropyl myristate
Adnan Sarwar Chaudhary
shown not to have antimicrobial activity in the
conditions of the test. Allow the oil to penetrate the
membrane by its own weight, and then filter,
applying the pressure or suction gradually. Wash
the membrane at least three times by filtering
through it each time about 100 ml of a suitable
sterile solution such as Fluid A (see Diluting and
Rinsing Fluids for Membrane Filtration) containing
a suitable emulsifying agent at a concentration
shown to be appropriate in the validation of the test,
for example polysorbate 80 at a concentration of 10
g per L (Fluid K). Transfer the membrane or
membranes to the culture medium or media, or vice
versa, as described above for Aqueous Solutions,
and incubate at the same temperatures and for the
same times.
OINTMENTS AND CREAMS
Use for each medium not less than the quantities of
the product prescribed in Tables 2 and 3. Ointments
in a fatty base and emulsions of the water-in-oil
type may be diluted to 1% in isopropyl myristate as
described above, by heating, if necessary, to not
more than 40. In exceptional cases it may be
necessary to heat to not more than 44. Filter as
rapidly as possible, and proceed as described above
for Oils and Oily Solutions.
PREFILLED SYRINGES
For prefilled syringes without attached sterile
needles, expel the contents of each syringe into one
or two separate membrane filter funnels or into
separate pooling vessels prior to transfer. If a
separate sterile needle is attached, directly expel the
syringe contents as indicated above, and proceed as
directed for Aqueous Solutions. Test the sterility of
the needle, using Direct Inoculation under
Validation Test.
SOLIDS FOR INJECTION OTHER THAN
ANTIBIOTIC
Constitute the test articles as directed on the label,
and proceed as directed for Aqueous Solutions or
Oils and Oily Solutions, whichever applies. [NOTE—
If necessary, excess diluent can be added to aid in
the constitution and filtration of the constituted test
article.]
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ANTIBIOTIC SOLIDS FOR INJECTION
Pharmacy Bulk Packages, < 5 g— From each of 20
containers, aseptically transfer about 300 mg of
solids, into a sterile 500-mL conical flask, dissolve
in about 200 mL of Fluid A (see Diluting and
Rinsing Fluids for Membrane Filtration), and mix;
or constitute, as directed in the labeling, each of 20
containers and transfer a quantity of liquid or
suspension, equivalent to about 300 mg of solids,
into a sterile 500-mL conical flask, dissolve in
about 200 mL of Fluid A, and mix. Proceed as
directed for Aqueous Solutions or Oils and Oily
Solutions, whichever applies.
Pharmacy Bulk Packages, 5 g— From each of 6
containers, aseptically transfer about 1 g of solids
into a sterile 500-mL conical flask, dissolve in
about 200 mL of Fluid A, and mix; or constitute, as
directed in the labeling, each of 6 containers and
transfer a quantity of liquid, equivalent to about 1 g
of solids, into a sterile 500-mL conical flask,
dissolve in about 200 mL of Fluid A, and mix.
Proceed as directed for Aqueous Solutions.
ANTIBIOTIC SOLIDS, BULKS, AND BLENDS
Aseptically remove a sufficient quantity of solids
from the appropriate amount of containers (see
Table 2), mix to obtain a composite, equivalent to
about 6 g of solids, and transfer to a sterile 500- mL
conical flask. Dissolve in about 200 mL of Fluid A,
and mix. Proceed as directed for Aqueous Solutions.
STERILE AEROSOL PRODUCTS
For fluid products in pressurized aerosol form,
freeze the containers in an alcohol-dry ice mixture
at least at –20 for about 1 hour. If feasible, allow the
propellant to escape before aseptically opening the
container, and transfer the contents to a sterile
pooling vessel. Add 100 mL of Fluid D to the
pooling vessel, and mix gently. Proceed as directed
for Aqueous Solutions or Oils and Oily Solutions,
whichever applies.
DEVICES WITH PATHWAYS LABELED
STERILE
Aseptically pass not less than 10 pathway volumes
of Fluid D through each device tested. Collect
the fluids in an appropriate sterile vessel, and
Adnan Sarwar Chaudhary
proceed as directed for Aqueous Solutions or Oils
and Oily Solutions, whichever applies.
In the case of sterile, empty syringes, draw sterile
diluent into the barrel through the sterile needle,
if attached, or through a sterile needle attached for
the purpose of the test, and express the
contents into a sterile pooling vessel. Proceed as
directed above. Direct Inoculation of the Culture
Medium Transfer the quantity of the preparation to
be examined prescribed in Tables 2 and 3 directly
into the culture medium so that the volume of the
product is not more than 10% of the volume of the
medium, unless otherwise prescribed.
If the product to be examined has antimicrobial
activity, carry out the test after neutralizing this
with a suitable neutralizing substance or by dilution
in a sufficient quantity of culture medium. When it
is necessary to use a large volume of the product, it
may be preferable to use a concentrated culture
medium prepared in such a way that it takes into
account the subsequent dilution. Where
appropriate, the concentrated medium may be added
directly to the product in its container.
OILY LIQUIDS
Use media to which have been added a suitable
emulsifying agent at a concentration shown to be
appropriate in the validation of the test, for example
polysorbate 80 at a concentration of 10 g per
L.
OINTMENTS AND CREAMS
Prepare by diluting to about 1 in 10 by emulsifying
with the chosen emulsifying agent in a suitable
sterile diluent such as Fluid A (see Diluting and
Rinsing Fluids for Membrane Filtration).
Transfer the diluted product to a medium not
containing an emulsifying agent.
Incubate the inoculated media for not less than 14
days. Observe the cultures several times during
the incubation period. Shake cultures containing
oily products gently each day. However, when
thioglycollate medium or other similar medium is
used for the detection of anaerobic
microorganisms, keep shaking or mixing to a
minimum in order to maintain anaerobic conditions.
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CATGUT AND OTHER SURGICAL SUTURES
FOR VETERINARIAN USE
Use for each medium not less than the quantities of
the product prescribed in Tables 2 and 3.
Open the sealed package using aseptic precautions,
and remove three sections of the strand for
each culture medium. Carry out the test on three
sections, each 30-cm long, which have been cut
off from the beginning, the center, and the end of
the strand. Use whole strands from freshly
opened cassette packs. Transfer each section of the
strand to the selected medium. Use sufficient
medium to cover adequately the material to be
tested (20 mL to 150 mL).
SOLIDS
Transfer a quantity of the product in the form of a
dry solid (or prepare a suspension of the product
by adding sterile diluent to the immediate
container), corresponding to not less than the
quantity indicated in Tables 2 and 3. Transfer the
material so obtained to 200 mL of Fluid
Thioglycollate Medium, and mix. Similarly, transfer
the same quantity to 200 mL of Soybean–Casein
Digest Medium, and mix. Proceed as directed
above.
Adnan Sarwar Chaudhary
PURIFIED COTTON, GAUZE, SURGICAL
DRESSINGS, AND RELATED ARTICLES
From each package of cotton, rolled gauze bandage,
or large surgical dressings being tested,
aseptically remove two or more portions of 100- to
500-mg each from the innermost part of the
sample. From individually packaged, single-use
materials, aseptically remove the entire article.
Immerse the portions or article in each medium, and
proceed as directed above.
STERILE DEVICES
Articles can be immersed intact or disassembled. To
ensure that device pathways are also in
contact with the media, immerse the appropriate
number of units per medium in a volume of medium
sufficient to immerse the device completely, and
proceed as directed above. For extremely large
devices, immerse those portions of the device that
are to come into contact with the patient in a
volume of medium sufficient to achieve complete
immersion of those portions. For catheters where
the inside lumen and outside are required to be
sterile, either cut them into pieces such that the
medium is in contact with the entire lumen or fill
the lumen with medium, and then immerse the
intact unit.
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Leaker’s test A leak is a way for fluid to escape a container or

fluid-containing system, through which the contents
of container can escape or outside matter can enter
(Remington) the container.
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1. Ampoules are leakers.
2. Leaking through tiny holes or cracks.
3. To check the leakage from ampoules only.
Types:
Sealing is of two types:
1. Tip sealing (heating until tip seal)
They are made by melting sufficient glass at the tip
of the ampoule neck to form a bead of the glass and
close the opening.
2. Pull sealing (pull the red hot ampoule to seal)
They are made by heating the neck of a rotating
ampoule below the tip, then pulling the tip away to
form a small twisted capillary just prior to being
melted closed.
Purpose:
1. To check the incompletely sealed ampoules
so that they may be discarded.
2. Can cause the interchange between the
content of ampoules and environment.
3. Microorganisms and other contaminants can
enter the ampoules and deteriorate it or
content release outside and spoil the
packaging.
Procedure:
1. This test is performed by producing negative
pressure within an incompletely sealed
ampoule while the ampoule is submerged
entirely in a deeply dye solution.
2. Most often, approximately 1% methylene
blue solution is employed.
3. After carefully rinsing the dye solution from
the outside, colour from the dye will be
visible within a leaker.
4. Leakers of course, are discarded.
Disadvantages:
1. Not for vials and bottles.
2. Capillaries of about 15 µm may or may not
be detected.
Adnan Sarwar Chaudhary
Clarity test
(USP)
Particulate matter in injections and parenteral
infusions consists of mobile undissolved particles,
other than gas bubbles, unintentionally present in
the solutions.
There are two types of clarity test depend upon the
type of particles present:
1. Subvisible particles
2. Visible particles
1. Subvisible particles
For the determination of particulate matter, two
procedures are as follows:
i. Method 1 (Light Obscuration Particle Count
Test)
ii. Method 2 (Microscopic Particle Count Test)
When Method 1 is not applicable, e.g., in the case
of preparations having reduced clarity or increase
viscosity, the test should be carried out according to
Method 2. Emulsions, colloids, and liposomal
preparations are examples.
Similarly, products that produce air or gas bubbles
when drawn into the sensor may also require
microscopic particle count testing.
i. Method 1 (Light Obscuration
Particle Count Test)
Principle:
Based on the principle of light blockage that allows
for an automatic determination of the size of
particles and the number of particles according to
size.
Apparatus:
An electronic liquid borne particle counting system
that uses a light obstruction sensor with a suitable
feeding device is used.
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Precautions: The preparation complies with the test if the

 The test is carried out in a laminar flow cabinet average number of particles present in the units
(Class 100 area). tested does not exceed 6000 per container equal to
 Very carefully wash the glassware and filtration or greater than 10 µm and does not exceed 600 per
equipment used, except for the membrane container equal to or greater than 25 µm.
filters, with a warm detergent solution, and rinse
Disadvantage:
with abundant amounts of water to remove all
Not used for viscous materials.
traces of detergent.
 Immediately before use, rinse the equipment ii. Method 2 (Microscopic Particle
from top to bottom, outside and then inside,
with particle-free water. Count Test)
Use a suitable binocular microscope, a filter
Procedure: assembly for retaining particulate matter, and a
1. Mix the contents of the sample by slowly membrane filter for examination.
inverting the container 20 times successively.
2. If necessary, cautiously remove the sealing Binocular Microscope
closure. a. Ocular micrometer (circular diameter
3. Clean the outer surfaces of the container graticule)
opening using a jet of particle-free water and
The large circle divided by crosshairs into quadrants
remove the closure, avoiding any
is designated the graticule field of view (GFOV).
contamination of the contents.
4. Eliminate gas bubbles by allowing to stand for Transparent and black circles having 10-µm and 25-
2 minutes or sonicating. µm diameters at 100× are provided as comparison
5. For large-volume parenterals, single units are scales for particle sizing.
tested.
6. For small-volume parenterals less than 25 mL A relative error of the linear scale of the graticule
in volume, the contents of 10 or more units are within ±2% is acceptable.
combined in a cleaned container to obtain a
volume of not less than 25 mL
7. Remove four portions of sample, not less than
5 mL each, and count the number of particles
equal to or greater than 10 µm and 25 µm.
8. Disregard the result obtained for the first
portion, and calculate the mean number of
particles for the preparation to be examined.

Acceptance Criteria:
Test-1.A (more than 100 mL) (25/ml =
10µm, 3/ml = 25 µm)

The preparation complies with the test if the

average number of particles present in the units
tested does not exceed 25 per mL equal to or greater
than 10 µm and does not exceed 3 per mL equal to
or greater than 25 µm.
Test-1.B (less than 100 mL) (6000/container = 10
µm, 600/container = 25 µm)
b. Stage micrometer
Ocular micrometer is calibrated using a stage
micrometer that is certified by either a domestic or
international standard institution. The large circle is
designated the graticule field of view (GFOV).

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c. Mechanical stage
It hold the entire filteration area of the membrane
filter
d. Illuminators
Two illuminators are required. One is an episcopic
brightfield illuminator internal to the microscope,
the other is an external, focusable auxiliary
illuminator that can be adjusted to give reflected
oblique illumination at an angle of 10 to 20.
Filter Assembly
The filter assembly for retaining particulate matter
consists of a filter holder made of glass or other
suitable material, and is equipped with a vacuum
source and a suitable membrane filter.
The membrane filter is of suitable size, black or
dark gray in color, nongridded or gridded, and 1.0
µm or finer in nominal pore size.
Procedure:
1.Mix the contents of the sample by slowly
inverting the container 20 times successively.
2.If necessary, cautiously remove the sealing
closure.
3.Clean the outer surfaces of the container opening
using a jet of particle-free water and remove the
closure, avoiding any contamination of the
contents.
4.Eliminate gas bubbles by allowing to stand for 2
minutes or sonicating.
5.For large-volume parenterals, single units are
tested.
6.For small-volume parenterals less than 25 mL in
volume, the contents of 10 or more units are
combined in a cleaned container to obtain a
volume of not less than 25 mL
7.Wet the inside of the filter holder fitted with the
membrane filter with several mL of particle-free
water. Filter all the solution.
8.Place the membrane filter in a Petri dish, and
allow the membrane filter to air-dry.
9.After the membrane filter has been dried, place
the Petri dish on the stage of the microscope,
scan the entire membrane filter under the
reflected light from the illuminating device, and
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count the number of particles that are equal to or
greater than 10 µm and the number of particles
that are equal to or greater than 25 µm.
10. Alternatively, partial membrane filter count
and determination of the total filter count by
calculation is allowed. Calculate the mean
number of particles for the preparation to be
examined.
Acceptance Criteria
Test-2.A (more than 100 mL) (12/ml = 10µm, 2/ml
= 25 µm)
The preparation complies with the test if the
average number of particles present in the units
tested does not exceed 12 per mL equal to or greater
than 10 µm and does not exceed 2 per mL equal to
or greater than 25 µm.
Test-1.B (less than 100 mL) (3000/container = 10
µm, 300/container = 25 µm)
The preparation complies with the test if the
average number of particles present in the units
tested does not exceed 3000 per container equal to
or greater than 10 µm and does not exceed 300 per
container equal to or greater than 25 µm.
Visible Test (visual
assessment of parenterals)
Principle
Visual assessment of parenterals
Apparatus:
1. Viewing station
2. Black panel (vertical position)
3. White panel (vertical position to the black panel)
4. Adjustable lamp holder (fitted with shaded white
light source)
The intensity of illumination at the viewing point is
maintained between 2000 lux and 3750 lux.
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Procedure:
1. Remove label from the bottle.
2. Wash the bottle from outside and dry it.
3. Invert, shake and swirl (rounding) to remove
the gas bubbles not so hard otherwise
bubbles are produced and observe for about
5 sec in front of white panel.
4. Repeat the procedure in front of the black
panel.
5. Record the presence of any particles.
Acceptance Criteria:
If particles are seen than reject otherwise accept.
Pyrogen Test
(USP 30 – NF 25)
Pyrogens are fever producing substances released
from either bacteria or viruses.
Pyro means ‗pyrexia‘
Gen means ‗producing‘
Two types are there:
1. Invivo Pyrogen Test (Rabbit test)
2. Invitro Pyrogen Test (LAL test)
The pyrogen test is designed to limit to an
acceptable level the risks of febrile reaction in the
patient to the administration, by injection, of the
product concerned. The test involves measuring the
rise in temperature of rabbits following the
intravenous injection of a test solution and is
designed for products that can be tolerated by the
test rabbit in a dose not to exceed 10 mL per kg
injected intravenously within a period of not more
than 10 minutes. For products that require
preliminary preparation or are subject to special
conditions of administration, follow the additional
directions given in the individual monograph or, in
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the case of antibiotics or biologics, the additional
directions given in the federal regulations.
APPARATUS AND DILUENTS
Render the syringes, needles, and glassware
free from pyrogens by heating at 250 for not less
than 30 minutes or by any other suitable method.
Treat all diluents and solutions for washing and
rinsing of devices or parenteral injection assemblies
in manner that will assure that they are sterile and
pyrogen-free. Periodically perform control pyrogen
tests on representative portions of the diluents and
solutions for washing or rinsing of the apparatus.
Where Sodium Chloride Injection is specified as a
diluent, use Injection containing 0.9 percent of
NaCl.
TEMPERATURE RECORDING
Use an accurate temperature-sensing device
such as a clinical thermometer, or thermistor probes
or similar probes that have been calibrated to assure
an accuracy of ±0.1 and have been tested to
determine that a maximum reading is reached in
less than 5 minutes. Insert the temperature sensing
probe into the rectum of the test rabbit to a depth of
not less than 7.5 cm, and, after a period of time not
less than that previously determined as sufficient,
record the rabbit's body temperature.
TEST ANIMALS
Use healthy, mature rabbits. House the
rabbits individually in an area of uniform
temperature between 20 and 23 and free from
disturbances likely to excite them. The temperature
varies not more than ±3 from the selected
temperature. Before using a rabbit for the first time
in a pyrogen test, condition it not more than seven
days before use by a sham test that includes all of
the steps as directed for Procedure except injection.
Do not use a rabbit for pyrogen testing more
frequently than once every 48 hours, nor prior to 2
weeks following a maximum rise of its temperature
of 0.6 or more while being subjected to the pyrogen
test, or following its having been given a test
specimen that was adjudged pyrogenic.
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PROCEDURE
Perform the test in a separate area
designated solely for pyrogen testing and under
environmental conditions similar to those under
which the animals are housed and free from
disturbances likely to excite them. Withhold all
food from the rabbits used during the period of the
test. Access to water is allowed at all times, but may
be restricted during the test. If rectal temperature-
measuring probes remain inserted throughout the
testing period, restrain the rabbits with light-fitting
neck stocks that allow the rabbits to assume a
natural resting posture. Not more than 30 minutes
prior to the injection of the test dose, determine the
―control temperature‖ of each rabbit: this is the base
for the determination of any temperature increase
resulting from the injection of a test solution. In any
one group of test rabbits, use only those rabbits
whose control temperatures do not vary by more
than 1 from each other, and do not use any rabbit
having a temperature exceeding 39.8 . Unless
otherwise specified in the individual monograph,
inject into an ear vein of each of three
rabbits 10 mL of the test solution per kg of body
weight, completing each injection within 10
minutes after start of administration. The test
solution is either the product, constituted if
necessary as directed in the labeling, or the material
under test treated as directed in the individual
monograph and injected in the dose specified
therein. For pyrogen testing of devices or injection
assemblies, use washings or rinsings of the surfaces
that come in contact with the parenterally
administered material or with the injection site or
internal tissues of the patient. Assure that all test
solutions are protected from contamination. Perform
the injection after warming the test solution to
a temperature of 37 ± 2. Record the temperature at
30-minute intervals between 1 and 3 hours
subsequent to the injection.
TEST INTERPRETATION AND
CONTINUATION
(USP30-NF25)
Adnan Sarwar Chaudhary
Consider any temperature decreases as zero
rise. If no rabbit shows an individual rise in
temperature of 0.5 or more above its respective
control temperature, the product meets the
requirements for the absence of pyrogens. If any
rabbit shows an individual temperature rise of 0.5
or more, continue the test using five other rabbits. If
not more than three of the eight rabbits show
individual rises in temperature of 0.5 or more and if
the sum of the eight individual maximum
temperature rises does not exceed 3.3, the material
under examination meets the requirements for the
absence of pyrogens.
TEST FOR PYROGENS
(BP 2013)
The test consists of measuring the rise in
body temperature evoked in rabbits by the
intravenous injection of a sterile solution of the
substance to be examined.
Selection of animals
Use healthy, adult rabbits of either sex
weighing not less than 1.5 kg, fed a complete and
balanced diet not containing antibiotics, and not
showing loss of body mass during the week
preceding the test. A rabbit is not be used in a
pyrogen test:
(a) If it has been used in a negative pyrogen test
in the preceding 3 days, or
(b) If it has been used in the preceding 3 weeks
in a pyrogen test in which the substance
under examination failed to pass the test.
Animals' quarters
Keep the rabbits individually in a quiet area
with a uniform appropriate temperature. Withhold
food from the rabbits overnight and until the test is
completed; withhold water during the test. Carry out
the test in a quiet room where there is no risk of
disturbance exciting the animals and in which the
room temperature is within 3 °C of that of the
rabbits' living quarters, or in which the rabbits have
been kept for at least 18 h before the test.
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Materials
Glassware syringes and needles. Thoroughly
wash all glassware, syringes and needles with water
for injections and heat in a hot-air oven at 250 °C
for 30 min or at 200 °C for 1 h.
Retaining boxes
The retaining boxes for rabbits whose
temperature is being measured by an electrical
device are made in such a way that the animals are
retained only by loosely fitting neck-stocks; the rest
of the body remains relatively free so that the
rabbits may sit in a normal position. They are not
restrained by straps or other similar methods which
may harm the animal. The animals are put into the
boxes not less than 1 h before the first record of the
temperature and remain in them throughout the test.
Thermometers
Use a thermometer or electrical device
which indicates the temperature with a precision of
0.1 °C and insert into the rectum of the rabbit to a
depth of about 5 cm. The depth of insertion is
constant for any one rabbit in any one test. When an
electrical device is used it may be left in position
throughout the test.
Preliminary test
After selection of the animals, one to three
days before testing the product to be examined, treat
those animals that have not been used during the
previous 2 weeks by intravenous injection of 10 mL
per kilogram of body mass of a pyrogen-free 9 g/L
solution of sodium chloride R warmed to about 38.5
°C. Record the temperatures of the animals,
beginning at least 90 min before injection and
continuing for 3 h after the injection of the
solution. Any animal showing a temperature
variation greater than 0.6 °C is not used in the main
test.
Main test
Carry out the test using a group of three rabbits.
Adnan Sarwar Chaudhary
Preparation and injection of the product
Warm the liquid to be examined to
approximately 38.5 °C before the injection. The
product to be examined may be dissolved in, or
diluted with, a pyrogen-free 9 g/L solution of
sodium chloride R or another prescribed liquid.
Inject the solution slowly into the marginal vein of
the ear of each rabbit over a period not exceeding 4
min, unless otherwise prescribed in the monograph.
The amount of the product to be injected varies
according to the product to be examined and is
prescribed in the monograph. The volume injected
is not less than 0.5 mL per kilogram and not more
than 10 mL per kilogram of body mass.
Determination of the initial and maximum
temperatures
The "initial temperature" of each rabbit is
the mean of two temperature readings recorded for
that rabbit at an interval of 30 min in the 40 min
immediately preceding the injection of the product
to be examined. The "maximum temperature" of
each rabbit is the highest temperature recorded for
that rabbit in the 3 h after the injection. Record the
temperature of each rabbit at intervals of not more
than 30 min, beginning at least 90 min before the
injection of the product to be examined and
continuing 3 h after the injection. The difference
between the maximum temperature and the initial
temperature of each rabbit is taken to be its
response. When this difference is negative, the
result is counted as a zero response.
Rabbits showing a temperature variation
greater than 0.2 °C between two successive
readings in the determination of the initial
temperature are withdrawn from the test. In any one
test, only rabbits having initial temperatures which
do not differ from one another by more than 1 °C
are used. All rabbits having an initial temperature
higher than 39.8 °C or less than 38.0 °C are
withdrawn from the test.
Interpretation of results
Having carried out the test first on a group
of three rabbits, repeat if necessary on further
groups of three rabbits to a total of four groups,
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depending on the results obtained. If the summed  Take Sample and LAL reagent in test tube
response of the first group does not exceed the and incubate for 1 hour.
figure given in the second column of the Table  Gel or clot formed
2.6.8.-1, the substance passes the test. If the  Rotate 180 o or invert it.
summed response exceeds the figure given in the  If clot break then test is negative otherwise
second column of the table but does not exceed the positive.
figure given in the third column of the table, repeat
the test as indicated above. If the summed response b. Quantitative:
exceeds the figure given in the third column of the
 Make dilutions of sample other procedure is
table, the product fails the test. Rabbits used in a test
same as in qualitative.
for pyrogens where the mean rise in the rabbits'
 Check lamda at different wavelengths.
temperature has exceeded 1.2 °C are permanently
excluded for Parenteral and other sterile ii. Photometric technique:
preparations a. Turbidimetric technique

Invitro Pyrogen Test (LAL test) This technique is a photometric test to measure the
(EP) increase in turbidity. Based on the test principle
employed, this technique may be classified as being
This test is used to check bacterial endotoxin.
either the end- point-turbidimetric test or the
Principle kinetic-turbidimetric test.
Extract protenious in nature
The end-point-turbidimetric test is based on the
Nature quantitative relationship between the endotoxin
Containing enzyme and proteins concentration and the turbidity (absorbance or
Source transmission) of the reaction mixture at the end of
Limulus polyphemus (horse shoe crab) an incubation period.
Take heart of mature crab and get blood ameobocyte
cells by lysis so called Limulus Amebocyte Lysate test. The kinetic-turbidimetric test is a method to
measure either the time (onset time) needed for the
Temperature
reaction mixture to reach a predetermined
36 - 38 °C during incubation or centrifugation
absorbance or transmission, or the rate of turbidity
pH: 5-7 development.

Instrument & Equipment The test is carried out at the incubation temperature
Depyrogenated (wash with water for injection or heat in recommended by the lysate manufacturer (usually
hot air oven at 250 °C for 30 minutes. 37 ± 1 °C).
Techniques
b. Chromogenic technique
Gel clot technique:
This technique is used to measure the chromophore
Apparatus released from a suitable chromogenic peptide by the
Incubator 36-38 o C, Hot air oven 250 o C, Pipette. reaction of endotoxins with the lysate. Depending
on the test principle employed, this technique may
Precaution: be classified as being either the end- point-
Avoid agitation otherwise gel not formed chromogenic test or the kinetic-chromogenic test.
Types: The end-point-chromogenic test is based on the
a. Qualitative: quantitative relationship between the endotoxin

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concentration and the quantity of chromophore Test A:

released at the end of an incubation period.
The kinetic-chromogenic test measures either the
time (onset time) needed for the reaction mixture to
reach a predetermined absorbance, or the rate of
colour development.
The test is carried out at the incubation temperature
recommended by the lysate manufacturer (usually
37 ± 1 °C).
Assay for active Ingredient
This test is used to confirm that every unit should
contains same amount of drug or active ingredients
with little variation in the batch.
Principle:
The test of the uniformity of the content of single
dose preparations is based on the assay of individual
contents of active substance(s) of a number of
dosage units to determine whether the individual
contents are within the limits set with reference to
the average content of the sample.
 For tablets, powders for parenteral use,
ophthalmic inserts, suspension for injection.
 The preparation complies with the test if
each individual content is between 85-115%
of the Average content.
 The preparation fails to comply with the test
if more than 1 individual content is outside
85-115% of the average content or if 1
individual content is outside 75-125% of the
average content.
 If 1 individual content is outside the limit of
85-115% but within the limit of 75-125%,
determine the individual content of another
20 dosage units taken at random.
 The preparation complies with the test if
more than 1 individual contents of the 30
units is outside 85-115% of the average
content and none is outside the limit of 75-
125% of the average content.
Safety Test (Remington)
It is entirely possible for a parenteral product to
pass the routine Sterility test, Pyrogen test, and
Chemical analysis, and still cause unfavourable
reactions when injected.
So, a safety test in animal is essential to provide the
additional assurance that the product does not have
unexpected toxic properties.

Method:
 Using a suitable analytical method,
determine the individual contents of active
substance(s) of 10 dosage units taken at
random.
 Now apply the criteria of Test A, Test B or
Test C as specified in the monograph for any
specified dosage form.
 Use Test A for liquid dosage form given
below:

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BIOLOGICAL ASSAYS

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biological effect, as produced by the

ASSAYS standard.
An assay is an investigative (analytic)  The standards are internationally accepted
procedure in laboratory medicine, pharmacology, samples of drugs maintained and
environmental biology, and molecular biology for recommended by the Expert Committee of
qualitatively assessing or quantitatively measuring the Biological Standardization of W.H.O.
the presence or amount or the functional activity of They represent the fixed units of activity
a target entity (the analyte), which can be a drug or (definite weight of preparation) for drugs
biochemical substance or a cell in an organism or
organic sample PURPOSE OF BIOASSAYS:
 Measurement of the pharmacological
BIOASSAYS: activity of new or chemically undefined
substances
Bioassay (commonly used short hand for biological  Investigation of the function of endogenous
assay), or biological standardization is a type of mediators
scientific experiment.  Determination of the side-effect profile,
Biological assays (also called bioassays) are an including the degree of drug toxicity
integral part of the quality assessment required for  Measurement of the concentration of known
the manufacturing and marketing of many substances.
biological and some non biological drug products.  Assessing the amount of pollutants being
Bioassays commonly used for drug potency released by a particular source, such as
estimation can be distinguished from chemical tests wastewater or urban runoff.
by their reliance on a biological substrate (e.g.  Determining the specificity of certain
animals, living cells, or functional complexes of enzymes to certain substrates.
target receptors). Because of multiple operational
and biological factors arising from this reliance on DIFFERENCE BETWEEN
biology, they typically exhibit a greater variability CHEMICAL AND
than do chemically-based tests. BIOLOGICAL ASSAY:
Bioassays are typically conducted to measure the
CHEMICAL ASSAY:
effects of a substance on a living organism and are
 Less time consuming
essential in the development of new drugs and in
 Economical
monitoring environmental pollutants.
 More precise and accurate
Bioassay is a procedure by which the potency or  Determine the amount of specific compound
the nature of a substance is estimated by studying or structural moiety
its effects on living matter.  Less chance of error
 Less reliable
Bioassay is a procedure for the determination of the
concentration of a particular constitute of a mixture. BIOASSAY:
 More time consuming
Principle of Bioassay:  Expensive
 The basic principle of bioassay is to  Less precise and accurate
compare the test substance with the  Measures the actual biological activity
International Standard preparation of the  More chance of error
same and to find out how much test  More reliable
substance is required to produce the same

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TYPES OF BIOASSAYS
Bioassays are of two types:
1. Quantal
2. Graded
1. QUANTAL:
 A quantal assay involves an "all or none
response". For example: Insulin induced
hypoglycemic convulsive reaction or the
cardiac arrest caused by digitalis. The
response is either +ve or -ve, there is no
intermediate response e.g.—either
convulsion occurs or doesn't occur; similarly
is with cardiac arrest.
 In case of toxicity studies, the animal
receiving a dose of drug either dies or does
not die. Also, no intermediate response is
possible. This is also known as the "all or
none" response assay. The quantal method
though not precise is employed for bioassay
of substance in the following ways:
o Comparison of threshold response or
o Comparison of effective dose
(ED50) or median lethal dose
(LD50)
2. GRADED:
 Graded assays are based on the observation
that there is a proportionate increase in the
observed response following an increase in
the concentration or dose. The parameters
employed in such bioassays are based on the
nature of the effect the substance is expected
to produce.
 For example: contraction of smooth muscle
preparation for assaying histamine or the
study of blood pressure response in case of
adrenaline.
 A graded bioassay can be performed by
employing any of the below-mentioned
techniques. The choice of procedure
depends on:
1. The precision of the assay required
2. The quantity of the sample substance
available
Adnan Sarwar Chaudhary
3. The availability of the experimental
animals.
BIOASSAY METHODS:
1. Matching bioassay
2. Interpolation method
3. Bracketing method
4. Multiple point bioassay
(i.e three point, four point and six
point bioassay).
1.Matching Bioassay:
 Matching Bioassay: It is the simplest type of
the bioassay. In this type of bioassay,
response of the test substance taken first and
the observed response is tried to match with
the standard response.
 Several responses of the standard drug are
recorded till a close matching point to that of
the test substance is observed.
 A corresponding concentration is thus
calculated. This assay is applied when the
sample size is too small.
 Since the assay does not involve the
recording of concentration response curve,
the sensitivity of the preparation is not taken
into consideration. Therefore, precision and
accuracy is not very good.
2.Interpolation Method:
 Interpolation bioassay: Bioassays are
conducted by determining the amount of
preparation of unknown potency required to
produce a definite effect on suitable test
animals or organs or tissue under standard
conditions.
 This effect is compared with that of a
standard. Thus the amount of the test
substance required to produce the same
biological effect as a given quantity the unit
of a standard preparation is compared and
the potency of the unknown is expressed as
a % of that of the standard by employing a
simple formula.
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Drawback of Interpolation Method:
 Many times, a reliable result cannot be
obtained using this calculation. Therefore it
may be necessary to adopt more precise
methods of calculating potency based upon
observations of relative, but not necessarily
equal effects, likewise, statistical methods
may also be employed.
 The data (obtained from either of assay
techniques used) on which bioassay are
based may be classified as quantal or graded
response.
 Both these depend ultimately on plotting or
making assumption concerning the form of
DRC.
Specificity of Bioassay:
 An accuracy with in ± 20% of the true value
is good and a ±10% is excellent..
Bioassay of Antibiotics
Antibiotics
An antibiotic is a medicinal preparation containing
a significant quantity of a chemical substance that is
produced naturally by a microorganism or
artificially by synthesis that has the capacity to
inhibit or destroy the microorganisms.
Principle
The potency of an antibiotic is estimated by
comparing the inhibition of growth of sensitive
micro-organisms produced by known
concentrations of the antibiotic to be examined and
a reference substance.
Acceptance Criteria
Acceptable limits for the sample is 95-105% of
estimated potency.
Methods
There are two methods for Antibiotics Assay
I. Diffusion Method
II. Turbidimetric Method
Adnan Sarwar Chaudhary
I. Diffusion Method
Principle:
Potency of antibiotic is determined by measuring
the diameter of zones of microbial growth inhibition
that occur due to the diffusion of antibiotic sample
into semisolid medium.
Procedure
A. Preparation of Petri dish
1. Liquefy a suitable medium (Agar medium)
2. Inoculate it with suitable micro-organisms
sensitive to antibiotic examined.
3. Pour this inoculated medium into petri dish.
4. Layer of the medium will be of uniform
thickness i.e. 2-5mm.
5. Store the dishes so that no appreciable
growth or death of the micro-organisms
occurs.
B. Preparation of reference & sample solutions
1. Using the solvent and the buffer solution
indicated in Table, prepare solutions of the
reference substance and of the antibiotic to
be examined.
Types of diffusion method
1. Cavity/well method
2. Disc method
1. Cavity/well method
 Holes of 5-8 mm in diameter are bored in
medium with sterile borer.
 The solutions are placed in the holes by
micropipette sufficient to fill the hole (1-
10µg).
 Place petri dish for 1-4 hrs at RT or at 4℃.
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 Incubate at a suitable temperature for about
18 hrs.
 Measure the diameters of the areas of the
circular inhibition zones with accuracy.
 In order to assess the validity of the assay,
use not fewer than 3 doses of the reference
substance and 3 doses of the antibiotic to be
examined
2. Disc method
 Use sterile absorbent paper discs of suitable
quality.
 Impregnate the disks with reference or test
solutions.
 Place the disc on the surface of medium.
 Place petri dish for 1-4 hrs at RT or at 4℃.
 Incubate at a suitable temperature for about
18 hrs.
 Measure the diameters of the areas of the
circular inhibition zones with accuracy.
II. Turbidimetric Method
Principle:
 This method is based on inhibition of
microbial growth as indicated by
measurement of the turbidity (transmittance)
of suspensions of a suitable micro-organism
in a fluid medium to which have been added
graded amount of test compound.
 Changes in transmittance produced by the
test compound are compared with those
produced by reference material.
Procedure
 Prepare a suitable medium e.g. Agar broth.
 Inoculate the medium (9ml) with a
suspension of chosen micro-organism (1ml)
sensitive to the antibiotic to be examined.
 Using the solvent and the buffer solution
indicated in Table, prepare solutions of the
reference substance and of the antibiotic to
be examined.
 Distribute an equal volume of each solution
into two identical test tubes.
 Add equal volume of inoculated medium to
each tube.
Adnan Sarwar Chaudhary
 Prepare at the same time 2 control tubes
without antibiotic, both containing the
inoculated medium and to one of which is
added immediately 0.5 mL of formaldehyde.
These tubes are used to set the optical
apparatus used to measure the growth.
 Incubate all test tubes for 3-4 hrs at a
suitable temperature i.e. 37 ℃ in water bath.
 After incubation, stop the growth of micro-
organisms by adding 0.5ml of
formaldehyde.
 Determine the amount of growth by
measuring transmittance with SP.
 Compare the growth of micro-organisms in
test with reference.
 Simply check the clarity with formaldehyde
control test tube for 100% inhibition.
Acceptance Criteria:
Clarity of solution
Bioassay of Insulin Injection
The most prominent manifestation of insulin
activity, an abrupt decrease in blood glucose, was
the basis for biologic assay from the time of its first
clinical use. The procedure, although relatively
cumbersome, has the great merit of accurately
reflecting the effect on the diabetic patient. The
advent of practical yet sophisticated
physicochemical methods (e.g., liquid
chromatography) to measure insulin potency
quantitatively has resulted in a more accurate and
precise compendial test for insulin and insulin
products. However, the bioidentity of insulin and
insulin products cannot be assessed by these
methods. Thus, a qualitative test in rabbits is
included in this chapter, and its use is called for in
the appropriate monographs.
The Rabbit Blood Sugar
Method
Quantitative is used to determine the potency of
Insulin Reference Standards, for the validation of
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the stability of new insulin preparations, and to
determine the specific activities of insulin analogs.
RABBIT BLOOD SUGAR
METHOD—QUANTITATIVE
USP Reference Standards
1. USP Dextrose RS
2. USP Insulin RS
3. USP Insulin (Beef) RS.
4. USP Insulin Human RS
5. USP Insulin (Pork) RS.
Diluent
Prepare an aqueous solution containing 0.1% to
0.25% (w/v) of either cresol or phenol, 1.4% to
1.8% (w/v) of glycerin, and sufficient hydrochloric
acid to produce a pH between 2.5 and 3.5, unless
otherwise directed in the individual monograph.
Standard Stock Solution
Dissolve either a suitable quantity of accurately
weighed USP Insulin RS or a vial of lyophilized
USP Insulin RS of the appropriate species in
Diluent to make a Standard Stock Solution
containing 40 USP Insulin Units per mL and having
a pH between 2.5 and 3.5, unless otherwise directed
in the individual monograph. Store in a cold place,
protected from freezing, and use within 6 months.
Standard Solutions
Dilute portions of the Standard Stock Solution with
Diluent to make two solutions, one to contain 1.0
USP Insulin Unit per mL (Standard Solution 1), and
the other to contain 2.0 USP Insulin Units per mL
(Standard Solution 2).
Assay Stock Solution
Proceed as directed under Standard Stock Solution,
except to use a suitable quantity of the preparation
under test in place of USP Insulin RS. The Assay
Stock Solution contains about 40 USP Insulin Units
per mL.
Assay Solutions
Dilute portions of the Assay Stock Solution with
Diluent to make two dilutions of the preparation
under test, one of which may be expected, on the
basis of the assumed potency, to contain 1.0 USP
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Insulin Unit per mL (Assay Solution 1), and the
other to contain 2.0 USP Insulin Units per mL
(Assay Solution 2). In the case of neutral insulin
injection, adjust to a pH of 2.5 to 3.5prior to making
the dilutions.
Doses of the Solutions To Be Injected
Select on the basis of trial or experience the dose of
the dilutions to be injected, the volume of which
usually will be between 0.30 mL and 0.50 mL. For
each animal the volume of the Standard Solution is
the same as that of the Assay Solution.
Preparation of Animal
Select suitable, healthy rabbits each weighing not
less than 1.8 kg. Keep the rabbits in the laboratory
for not less than 1 week before use in the assay,
maintaining them on an adequate uniform diet, with
water available at all times.
Procedure
Divide the rabbits into four equal groups of
preferably not less than six rabbits each. On the
preceding day, approximately 20 hours before the
assay, provide each rabbit with an amount of food
that will be consumed within 6 hours. Follow the
same feeding schedule before each test day. During
the assay, withhold all food until after the final
blood specimen is taken. Handle the rabbits with
care in order to avoid undue excitement, and inject
subcutaneously the doses indicated in the following
design (see Table 1), the second injection being
made on the day after the first injection, or not more
than 1 week later. The time between the first and
second injection is the same for all rabbits.
Table 1
Group First Injection Second
Injection
1 Standard Assay Solution 1
Solution
2
2 Standard Assay Solution 2
Solution
1
3 Assay Solution 2 Standard
Solution
1
4 Assay Solution 1 Standard
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Solution values, and subtract its response, disregarding the

2 chronological order in which the responses were
observed, to obtain the individual differences, y, as
Blood Samples
shown in Table 2.
At 1 hour ± 5 minutes and 2½ hours ± 5 minutes
after the time of injection, obtain from each rabbit a When the data for one or more rabbits are missing
suitable blood specimen from a marginal ear vein. in an assay, do not use the confidence interval
Blood can also be collected effectively from the formulas given here, but seek statistical help. The
central auricular artery. data can still be analyzed with proper analysis of
variance. When the number of rabbits, f, carried
Dextrose Determination through the assay is the same in each group, total
the y's in each group and compute Ta = –T1 + T2 +
Determine the dextrose content of the blood T3 – T4 and Tb = T1 + T2 + T3 + T4. The
specimens by a suitable procedure that is adapted
logarithm of the relative potency of the test
to automated analysis. The following procedure
dilutions is M ′ = 0.301Ta / Tb. The potency of the
may be used.
injection in USP Units per mg equals the antilog
Anticoagulant Solution (log R + M ′), where R = vS / vU, in which vS is the
Dissolve 1 g of edetate sodium and 200 mg of number of USP Units per mL of the Standard
sodium fluoride in 1 L of water, and mix. dilution and vU is the number of mg of insulin per
mL of the corresponding Assay dilution.
Dextrose Standard Preparations
Determine the 95% confidence interval for the log-
Transfer known concentrations of USP Dextrose
relative potency using Fieller's Theorem. If the
RS to suitable vessels, and dilute quantitatively and
stepwise with Anticoagulant Solution (1:9) to obtain confidence interval is more than 0.082, which
a range of Dextrose Standard Preparations corresponds at P = 0.95 to confidence limits of
containing between 20 and 100 mg per 100 mL, about ±10% of the computed potency, repeat the
having known concentrations similar to the assay until the combined data of the two or more
concentrations in the rabbit blood samples. assays, redetermined as described in Combination
of Independent Assays under Design and Analysis of
Test Preparations
Biological Assays , meet this acceptable limit.
Pipet into separate, suitable vessels 0.1 mL of each
Blood Sample and 0.9 mL of Anticoagulant Table 2
Solution. Gro Differe Indivi Total Standar
up nces dual Respo d
Procedure Respo nse Deviati
Subject the Test Preparations to dialysis across a nse (T) ons of
semipermeable membrane for a sufficient time so (y) Differe
that the dextrose passes through the membrane into nces
a saline TS solution containing glucose oxidase, (S)
1 Standar y1 T1 S1
horseradish peroxidase, 3-methyl-2
d
benzothiazolinone hydrazine hydrochloride TS, and Solution
N,N-dimethylaniline. The absorbances of the Test 2 –
Preparations are determined at 600 nm in a Assay
recording colorimeter. The absorbances of the Solution
Dextrose Standard Preparations are similarly 1
2 Assay y2 T2 S2
determined at the start and the end of each run.
Solution
2 –
Calculation
Standar
Calculate the response of each rabbit to each d
injection from the sum of the two blood-sugar
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Solution Procedure:
1
3 Assay y3 T3 S3 1. Bioassay by Guinea Pig
Solution  Take 12 guinea pig, each weighing between
2 –
Standar 200-600g.
d  Weight of the heaviest and lightest animal
Solution should not differ > 100g.
1  Distribute the pigs at random into two
4 Standar y4 T4 S4
groups.
d
Solution  Mean body weights of the two groups
2 – should not differ by >10%.
Assay  Extract of both test and standard are diluted
Solution with normal saline (1g in 80ml).
1
 The jugular vein is traced out by removing
the adhering tissues and cannulated by
means of venous cannula.
Assay of prepared digitalis  A pin is inserted in the heart such that it gets
inserted in the apex of heart. In this way we
Introduction: can observe the heart beats by up and down
 Digitalis is Cardio-active glycoside. movements of the pin.
 Acts directly on myocardium and increase  Extract is injected at slow uniform rate into
force of contraction jugular vein of anesthetized animal between
 Used in CHF duration of 20-40 min.
 Injection is continued until heart is arrested.
Principle:  Volume of extract administered is taken as
Potency of test is compared with that of standard lethal dose.
preparation by determining the action on cardiac
muscles. Bioassay by Pigeons
Standard Preparation & Units  Take 12 pigeons and divided into 2 groups.
 It consists of a mixture of dried and  Weight of largest pigeon should not exceed
powdered digitalis leaves. twice the weight of smallest pigeon.
 Supplied in ampoules having 2.5g digitalis.  Mean body weights of the two groups
should not differ by >30%.
1 unit = 76 mg of standard  Extract of both test and standard are diluted
with normal saline (1g in 80ml).
0.01316 unit = 1 mg of standard
 Extract is injected at slow uniform rate into
Preparation of Extract alar vein of slightly anesthetized and
 Transfer the known quantity of powder in immobilized pigeon.
container approx. 50ml capacity.  Dose 1ml/kg is administered within few
 Add 10ml of alcohol (80%) for each gram of seconds and repeated at 5 min interval until
powder. heart is arrested.
 Close the container and shake continuously:  In pigeons, stoppage of heart is associated
24 23 o For 48 hours at 10-20 ℃ with a characteristic vomiting response
 Centrifuge and filter it called ‗Emesis‘ (the milk from the crop sac
 Store in dry and tightly closed bottle at 5 to - of pigeon is ejected out). This may take as
5℃ the end point response of the digitalis.
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 Lethal dose per kg is equal to number of than 60 g, or that shows evidence of injury,
doses received. disease, or anatomical abnormality.

Acceptance Criteria: 2. Depletion Period

Acceptable limits for the sample is 85-125% of  Depletion period, which extends from the
estimated potency. end of the preliminary period to the first day
of the assay period.
Assay of Vitamin D.  Provide each rat with the Rachitogenic Diet
and water
Vitamin-D
3. Rachitogenic Diet
1. Vitamin D is fat soluble vitamin.
2. It maintain the adequate plasma level mixture of the following ingredients in the
of calcium in the body. proportions shown in the accompanying table.
3. Deficiency results in rickets in
children and osteomalacia in adults

Principle Ingredient Parts by weight

The activity of a preparation of vitamin D is
Whole yellow corn, ground 76
determined by comparing its antirachitic activity
with that of standard preparation by a suitable Wheat gluten, ground 20
method
Calcium carbonate 3
 USP Reference Standard:
 USP Cholecalciferol RS Sodium chloride 1

Subject:
Very young rats (litters)
Assigning Rats to Groups for Assay Period
Procedure
 Consider a litter suitable for the assay period
1. Preliminary Period
when individual rats in the litter show
2. Depletion Period
evidence of rickets such as enlarged joints
3. Rachitogenic Diet
and a distinctive wobbly, rachitic gait,
4. Assigning Rats to Groups for Assay Period
provided that the depletion period is not less
5. Administration of Standard and Sample
than 19 or more than 25 days.
Doses
 The presence of rickets may be established
6. Assay Period
also from the width of the rachitic
7. Line Test
metaphysis upon X-ray examination or by
8. Observations and Acceptability
applying the Line Test (described below) to
Preliminary Period a leg bone of one member of each litter.
 30 days period and extends from birth to the  Record the weight of each rat, and assign it
first day of the depletion period to a group
 During the preliminary period, use a dietary  Litters are divided into 4 groups each
regimen that provides for normal containing 7 or more litters.
development but is limited in its content of
5. Administration of Standard and Sample Doses
vitamin D.
 The litters in two reference groups receive x
 At the end of the preliminary period, reject
and nx doses respectively.
any rat that weighs less than 44 g or more
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 The litters in two sample group receive the  Score the degree of calcification of the
dose in same ratio. rachitic metaphysis in each rat, according to
 Give the dose at once or in 8 divided doses. a scale that allows the average response to
 Ratio of larger to smaller dose not less than be plotted as a straight line against the
1.5 or more than 2.5. logarithm of the dose.
 Before feeding, the Reference Standard
and/or sample may be diluted with
cottonseed oil, provided that not more than
0.2 mL is fed on any one day.
 Store the oil solutions in well-closed bottles,
protected from light, at a temperature not
exceeding 10°, and use within 5 weeks.

6. Assay Period
 Assay period last for 7-10 days.
 Throughout the assay period, maintain as
uniform environmental conditions as
possible for all rats, and exclude exposure to
antirachitic radiations.
 At the end of a fixed period of 7 to 10 days,
weigh and kill each rat.
 Dissect out one or more leg bones for
examination by the Line Test.

7. Line Test
 Remove the proximal end of a tibia or the
distal end of a radius, and clean adhering
tissue from it, in any one assay using the
same bone from all animals.
 Rinse in purified water, immerse
immediately in silver nitrate solution (1 in
50) for 1 minute, and rinse again in purified
water.
 Expose the cut surface of bone, in water, to
daylight or another source of actinic light
until the calcified areas develop a clearly
defined stain without marked discoloration
of the uncalcified areas.
 The staining procedure may be modified to
differentiate more clearly between calcified
and uncalcified areas.

8. Observations And Acceptability

 Calcified and uncalcified areas are observed
and relative potency of standard and sample
is calculated.

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ALCOHOL
DETERMINATION

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“The measurement of percentage of alcohol in
different dosage forms.”
Properties of Alcohol
Alcohol means ‗ethanol‘, used in preparations
because of its important properties:
 Used as solvent
 As co-solvent along with water
 Used because of its preservative properties.
 Nontoxic in nature.
Methods To Determine
Alcohol
There are two methods for determination of alcohol:
1. Distillation method.
2. Gas Liquid Chromatography.
Method I (Distillation
Method)
Method I is to be used for determination of alcohol
unless otherwise specified in individual monograph.
It is suitable for examining fluid extracts, tinctures,
provided the capacity of the distilling flask is
sufficient (commonly 2 to 4 times the volume of the
liquid to be heated) and the rate of distillation is
such that clear distillates are produced.
Procedure:
Two types of procedures are there:
1. Normal Procedure
2. Special Procedure
Normal Procedure:
For liquids containing 30% alcohol or less
• By using pipette transfer liquid dosage form in
distilling apparatus equal to 25ml.
• Note temperature and add equal volume of
water
• Pour into distillation apparatus and start the
process
Adnan Sarwar Chaudhary
• Collect the volume of distillate about 2ml less
than volume of original test liquid
• Maintain the temperature as it was at the start
of process
• Add some quantity of water to make original
volume of test liquid
• Resulted distillate may be clear or not if not
then clear it by adding the talc or CaCO3
• Now determine the specific gravity at 25 oC
• So we can calculate the %age v/v of alcohol.
• The distillate is clear or not more than slightly
cloudy, and does not contain more than traces
of volatile substances other than alcohol and
water. If distillate is not clear than add talc or
calcium carbonate, and filtered, after which the
temperature of the filtrate is adjusted and the
alcohol content determined from the specific
gravity.
• Take precautions to minimize the loss of
alcohol by evaporation during all
manipulations.
For liquids containing more than 30% of alcohol
• Take 25ml of the liquid in which the alcohol is
to be determined
• Add twice volume of water in the sample
liquid.
• Subject this liquid to distillate.
• Collect a volume of distillate about 2ml less
than the twice volume of sample liquid taken
(about 48ml) and bring the temperature at
which original liquid was measured.
• Add sufficient water to measure exactly the
double volume of sample liquid taken i.e.50ml
• Determine the specific gravity of liquid.
• The proportion of ethanol, by volume, in this
distillate, as ascertained from its specific
gravity, equals one half that in the liquid
examined.
Special Procedure
For liquids containing 50% alcohol or less
• Take 25ml of the sample to be examined in
separating funnel.
• Mix an equal volume of water in it.
• Saturate this mixture with sodium chloride.
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• Add 25ml of solvent hexane, shake the mixture
to extract the interfering volatile ingredients.
• Draw off the lower layer into second separating
funnel.
• Repeat the extraction twice with two further
25ml portions of solvent hexane.
• Extract the combined solvent hexane solutions
with three 10ml portions of a saturated solution
of sodium chloride.
• Separate aqueous layer i.e. NaCl solution.
• Combine the saline solution and distill in usual
manner, collecting a volume of distillate having
a simple ratio to the volume of the original
sample. (via same procedure as for less than
30% alcohol)
• Collect the distillate and determine %age of
alcohol & specific gravity.
For liquids containing more than 50% alcohol
• Adjust the specimen under examination by
diluting it with water up to 25% ad then
performed process as above.
• In preparing collodion or flexible collodion, use
water in place of saturated NaCl solution as
directed above.
• The remaining procedure is same.
• If volatile oils are presents in small proportion
only, and a cloudy distillate is obtained, the
solvent hexane treatment not having been
employed. The distillate may be clarified by
shaking it with about one-fifth its volume of
solvent hexane, or by filtering it through a thin
layer of talc.
Special Consideration
a) Volatile Acids & Bases:
If preparations contain volatile bases make it
slightly acidic with diluted sulfuric acidic before
distilling. If volatile acids are present make the
preparation slightly alkaline with sodium
hydroxideTS.(4% NaOH)
b) Glycerin:
If sample contain glycerin add sufficient water so
that the residue, after distillation, contains not less
than 50% of water.
Adnan Sarwar Chaudhary
c) Iodine:
If the sample solution contain free iodine than treat
sample with powdered zinc before distillation.
OR
Decolorize with just sufficient sodium thiosulfate
solution (1in10), followed by a few drops of sodium
hydroxideTS.
d) Other Volatile Substances:
Spirits, elixirs , tinctures, and similar preparations
that contain appreciable proportions of volatile
materials other than alcohol and water, such as
volatile oils , chloroform , ether , camphor , etc.,
require special treatment, as follows:
Problems during Distillation of
Alcohol
Following problems occurred during distillation of
alcohol:
1. Rothing / Foam Formation:
 The foam formation in the distillation flask
is frothing due to presence of surface active
agents.
 It may cause problem in accurate measuring.
Remedy:
 Treat with slight excess of CaCl₂ solution.
 Addition of small amount of paraffin oil or
silicone oil before starting distillation.
2. Bumping:
 Bubble formation in distillation flask is
called bumping.
 It may exert pressure on the walls of vessel
which may break the vessel.
Remedy:
 It can be prevented by adding porous chips
of insoluble material e.g., silicon carbide,
glass chips, beads.
3. Cloudy / Milky Distillation: Cause problem
in measuring. Remedy
Clarification is done by adding talc or calcium
carbonate, chalk and then filter.
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4. Azeotrope Formation:
 Complex formation between two agents
which may change the nature of individual
components is called Azeotrope.
 Azeotropes are also called constant boiling
mixtures.
 They are binary mixtures having the same
composition in liquid and vapor phase and
boil at a constant temperature. In such a case
it is not possible to separate the components
by fractional distillation.
Example:
Water+ acetonitrile (Azeotrope formation)
5. Emulsification:
It can be problematic for the process of distillation.
Remedy:
 Distillate is saturated with brine and light
petroleum.
 Excessive temperature can break emulsion.
METHOD II (GAS LIQUID
CHROMATOGRAPHY)
USP Reference Standards
1. USP Alcohol Determination—Acetonitrile
RS
2. USP Alcohol Determination—Alcohol RS.
Method IIa
Apparatus
Gas chromatography specification
1. Flame-ionization detector
2. 4-mm × 1.8-m glass column packed with 100-
to 120-mesh chromatographic column packing
support S3
3. Nitrogen or helium as the carrier.
Temperature specification
1. Prior to use, condition the column overnight at
235oC with a slow flow of carrier gas.
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2. The column temperature is maintained at 120o
3. The injection port and detector temperatures
are maintained at 210o.
4. Adjust the carrier flow and temperature so that
acetonitrile, the internal standard, elutes in 5
to 10 minutes.
Solutions:
Test Stock Preparation:
Dilute the specimen under examination stepwise
with water to obtain a solution containing
approximately 2% (v/v) of alcohol.
Test Preparation:
Pipet 5 mL each of the Test Stock Preparation and
the USP Alcohol Determination—Acetonitrile RS
[Alternatively, a 2% aqueous solution of acetonitrile
of suitable quality may be used as the internal
standard solution] into a 50-mL volumetric flask,
dilute with water to volume, and mix.
Standard Preparation:
Pipet 5 mL each of the USP Alcohol
Determination—Alcohol RS and the USP Alcohol
Determination—
Acetonitrile RS [Alternatively, a 2% aqueous
solution of acetonitrile of suitable quality may be
used as the internal standard solution] into a 50-mL
volumetric flask, dilute with water to volume, and
mix.
Procedure
 Inject about 5 µL each of the Test
Preparation and the Standard Preparation, in
duplicate, into the gas chromatograph,
record the chromatograms, and determine
the peak response ratios.
 Calculate the percentage of alcohol (v/v) in
the specimen under test according to the
formula:
CD(RU / RS)
o C is the labeled concentration of USP
Alcohol Determination—Alcohol RS
o D is the dilution factor (the ratio of the
volume of the Test Stock Preparation to the
volume of the specimen taken);
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o RU and RS are the peak response ratios
obtained from the Test Preparation and the
Standard Preparation, respectively.
System Suitability Test
 In a suitable chromatogram, the resolution
factor, R, is not less than 2;
 The tailing factor of the alcohol peak is not
greater than 2.0
 Six replicate injections of the Standard
Preparation show a relative standard deviation
of not more than 2.0% in the ratio of the peak of
alcohol to the peak of the internal standard.
Method IIb
Apparatus:
Gas chromatography specification
1. Split injection port with a split ratio of 5:1
2. A flame-ionization detector
3. 0.53-mm × 30-m capillary column coated with a
3.0-µm film of phase G43.
4. Helium is used as the carrier gas at a linear
velocity of 34.0 cm per second.
Temperature specification
1. The chromatograph is programmed to maintain
the column temperature at 50 o for 5 minutes
2. Then to increase the temperature at a rate of 10 o
per minute to 200 o , and maintain at this
temperature for 4 minutes.
3. The injection port temperature is maintained at
210 o
4. The detector temperature at 280 o .
Solutions:
Test Stock Preparation:
Dilute the specimen under examination stepwise
with water to obtain a solution containing
approximately 2% (v/v) of alcohol.
Test Preparation:
Pipet 5 mL each of the Test Stock Preparation and
the USP Alcohol Determination—Acetonitrile RS
[Alternatively, a 2% aqueous solution of acetonitrile
of suitable quality may be used as the internal
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standard solution] into a 25-mL volumetric flask,
dilute with water to volume, and mix.
Standard Preparation:
Pipet 5 mL each of the USP Alcohol
Determination—Alcohol RS and the USP Alcohol
Determination— Acetonitrile RS [Alternatively, a
2% aqueous solution of acetonitrile of suitable
quality may be used as the internal standard
solution] into a 25-mL volumetric flask, dilute with
water to volume, and mix.
Procedure:
 Inject about 0.2 to 0.5 µL each of the Test
Preparation and the Standard Preparation, in
duplicate, into the gas chromatograph,
record the chromatograms, and determine
the peak response ratios.
 Calculate the percentage of alcohol (v/v) in
the specimen under test according to the
formula:
CD(R U / R S )
o C is the labeled concentration of
USP Alcohol Determination—
Alcohol RS
o D is the dilution factor (the ratio of
the volume of the Test Stock
Preparation to the volume of the
specimen taken);
o R U and R S are the peak response
ratios obtained from the Test
Preparation and the Standard
Preparation, respectively.
System Suitability Test:
 In a suitable chromatogram, the resolution
factor, R, between alcohol and the internal
standard is not less than 4
 The tailing factor of the alcohol peak is not
greater than 2.0
 Six replicate injections of the Standard
Preparation show a relative standard
deviation of not more than 4.0% in the ratio
of the peak of alcohol to the peak of the
internal standard.Used only when specified
in individual monograph.
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ALKALOIDAL
DRUG ASSAY

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Alkaloids
Alkaloids are the organic compounds normally with
basic chemical properties and usually containing at
least one nitrogen atom in a heterocyclic ring,
occurring chiefly in many vascular plants and some
fungi; having marked physiological effects on
humans or animals. Many alkaloids, such as
nicotine, quinine, cocaine, and morphine, are known
for their poisonous or medicinal attributes.
Assay
The determination of the activity, potency, strength,
etc. of a substance, either on an absolute basis or in
comparison with that of a standard preparation.
OR
Qualitative or quantitative analysis of a substance,
especially of a drug, to determine its components.
OR
Determination of the amount of a particular
constituent of a mixture, or of the potency of a drug.
ALKALOIDAL DRUG
ASSAY
The assay of the drugs containing alkaloids is called
―Alkaloidal Drug Assay‖.
Examples
2. Morphine sulphate
3. Epinephrine
4. Hyoscyamine
5. Atropine
6. Papaverine
7. Caffeine
Introduction
 Alkaloids are slightly or very slightly
soluble in water.
 Alkaloids are soluble in certain organic
solvents immiscible with water e.g.
chloroform.
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 Salts of alkaloids are soluble in water but
almost insoluble in organic solvents.
 The process of assay is carried out by
treating the drug with a solvent immiscible
with water in the presence of excess of alkali
that liberates the alkaloid.
 The free alkaloid is dissolved by the
immiscible solvent from which it is removed
by means of excess of dilute acid.
 The acid solution is then extracted with an
immiscible solvent in the presence of alkali.
 The immiscible solvent is then evaporated to
obtain the alkaloid which is either weighed
or determined volumetrically.
Preparation of Drug for Assay

powder of fineness grade.

water.
Weighing for Assay
a. Weighing of bulky crude drugs
In weighing bulky crude drugs for the assay,
accuracy to within 10 mg for quantities of 5 gm and
over is sufficient.
b. Portions of soft extracts or ointments
These may be weighed on a tarred piece of wax
paper and transferred to the vessel containing the
solvent for extraction.
Extraction of drugs
The alkaloidal content of alkaloid-bearing drugs is
usually extracted by one of the following methods.
1. Maceration
2. Percolation
3. Continuous Extraction
1. Maceration
Introduction
 The term maceration comes from the Latin
‗macerare‘ which means ‗to soak‘.
 It is a process in which the properly crushed
drug is permitted to soak in the menstrum
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until the cellular structure is softened and
penetrated by the menstrum.
 By this method, nearly all the soluble
contents are dissolved in the menstrum.
Menstrum: A solvent, especially one used in
extracting compounds from plant and animal tissues
and preparing drugs.
 Maceration is different from water based
infusions and decoctions in following
respects:
o The menstrum is usually alcohol.
o The herb remains in the menstruum for a
longer period of time.
o The process is conducted at ordinary
temperature.
Procedure
Method 1
 The drug to be extracted is placed in a wide-
mouth container containing certain
menstruum.
 The vessel is closed tightly to prevent the
loss of menstruum.
 The contents are shaken/agitated repeated
(preferably on daily basis) for a period of 2
to 14 days.
 The agitation allows the repeated flow of
fresh menstrum over the entire surface area
of soaked comminuted drug.
 After the specific time, the liquid is drained
from the mark. The mark is then pressed to
retrieve more of the menstruum.
 The expressed liquid is mixed with the
strained liquid and the mixture is left to
stand until it is clear, after which it is
filtered.
Method 2
 Place the drug in a porous cloth bag that is
tied and suspended in the upper portion of
menstruum. (Just like a teabag).
 As the soluble contents dissolve in the
menstruum, they tend to settle to the bottom
because of an increase in the specific gravity
of the liquid.
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 The extractive is separated from the marc by
expressing the bag of drug and washing it
with additional menstruum.
 The menstruum is then filtered.
Specifications for Maceration
 Maceration is usually conducted at a
temperature between 15 to 20 ° C.
 Duration is from minimum 2 days to
maximum 14 days.
 Wide mouth vessel is used.
 Mouth of vessel is stoppered tightly.
 Contents are agitated repeatedly.
Examples:
 Benzoin
 Aloe
 Tolu
Percolation
Introduction
The term percolation is derived from the Latin ‗per‘
meaning ‗through‘ and ‗colare‘ meaning ‗to strain‘.
So
―It is a process in which a comminuted drug is
extracted of its soluble constituents by a slow
passage of a suitable solvent through a column of
the drug‖.
According to USP
―Percolation consists in subjecting a comminuted
substance or a mixture of substances contained in a
vessel called a percolator, to the solvent action of a
liquid termed as menstruum in such a manner that
the liquid shall extract the soluble constituents and
pass from the percolator‖.
Procedure
 The comminuted drug is packed in a special
extraction apparatus termed as percolator by
packingNthe outlet with some porous
material e.g. cotton.
 Saturate the drug with specified solvent and
allow standing for 5 minutes.
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 Add some ammonia sufficient to make the

mixture distinctly alkaline and mix
thoroughly with drug.
 From this percolator, the suitable
solvent(menstruum) is passed through
slowly which dissolves the soluble
constituents of the comminuted drug.
 Allow the drug to macerate for about 10 to
12 hours.
 That menstruum is now called ―Percolate‖
and is collected from the bottom of
percolator until the drug is completely
exhausted of its alkaloidal content.
 The flow of the menstruum is generally
downwards due to gravity.
 In some specialized and more sophisticated
percolation apparatus, additional pressure is
exerted on the column with positive air
pressure at the inlet and suction at the outlet.

Determination of the completeness of extraction of

alkaloid
 Take about 4 ml of the last percolate.
 Evaporate it to dryness.
 Dissolve the residue in 0.5 ml of 0.5 N acid.
 Add a drop of mercuric iodide(Valser‘s
Reagent)
 A slight turbidity is produced.

Percolators
 Percolators employed on large scale
industrial preparations are generally made
up of stainless steel or glass lined metal
vessels and vary greatly in size & operation.
 For example percolators used to extract from
leaves may be 6 to 8 feet in diameter and 12
to 18 feet high. Continuous Extraction
 Percolators used on small scale are usually Procedure
made up of glass.
 Moisten the drug with a specified solvent.
Shapes  Allow to stand for 5 mins.
 Cylindrical  Make the mixture alkaline using ammonia
 Roundish and mix thoroughly.
 Conical  Allow the drug to macerate for 6-12 hrs.
 Funnel shaped  Then pack the drug in the thimble and cover
it with a pledge of cotton.

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 Take the thimble and insert it into a suitable
extractor e.g. Soxhlet extractor.
 Add sufficient quantity of solvent and
extract the drug.
PURIFICATION OF
ALKALOIDS
 The alkaloidal solution obtained by any of
the extraction methods is usually
contaminated with other extractives which
interfere with the quantitative determination
of alkaloids.
 For effective purification, remove the
alkaloids from the immiscible solvent by
shaking out with an acid.
 Then make the acidic solution alkaline with
an alkali hydroxide and extract with an
immiscible solvent.
 The volume & strength of the acid vary case
to case. However total volume should be as
small as possible.
 Shake the combined acid extracts with one
or more 10 ml portions of the appropriate
immiscible solvent until the acid solution is
clear. Then wash the immiscible solvent
Adnan Sarwar Chaudhary
extracts with one or more 5 ml portion of
water acidified with HCl or H 2 SO 4 and
add these washings to the acid solution.
 Then make the acid solution alkaline with
ammonia and extract it with some
immiscible solvent. Repeat the operation as
long as any alkaloid is extracted by the
immiscible solvent. The completeness of the
extraction can be tested by mercuric iodide
TS.
 In all assays, continues the extraction until
0.5 ml of the last acid washing shows a very
slight turbidity on the addition of a drop of
mercuric iodide.
DETERMINATION OF
ALKALOIDS
 Evaporate the solution of the purified
alkaloids in the immiscible solvent to
dryness on a steam bath or with a current of
air.
 Soften the alkaloidal residue by addition of
about 1 ml of the neutralized alcohol or
ether.
 Add an accurately measured volume of
standard acid.
 Warm the mixture gently.
 Dissolve the alkaloidal residue in
chloroform and add standard acid of higher
normality.
 Remove the chloroform completely by
evaporation.
 The add water (q.s) to make the volume of
mixture at least 25 ml.
 Titrate the excess of the acid with standard
alkali.
 Dry the alkaloidal residue at 105 °C to a
constant weight.
ESTIMATION OF ALKALOIDS
Following tests are used to detect the presence of
alkaloids:
1. Mayer’s Test:
It gives white or yellow ppt except with alkaloids of
purine group.
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2. Dragendroff’s Test 6. Melting Range Test

Orange colour ppt formed. Every alkaloid has specific melting range like
Atropine has 114-118 o C.
3. Wagner’s Test
Brown or reddish brown ppt.

4. Hagger’s Test
Characteristics crystalline ppt.

5. Tannic Acid Test

Freshly prepared tannic acid solution gives ppt
which is insoluble in dilute acid.

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QUALITY
ASSURANCE OF
VACCINES

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Immunological Products
Group of pharmaceutical preparations with diverse
origins but a common pharmacological purpose:
Modification of the Immune status of a recipient,
either to provide immunity to infectious diseases or
to aid in the detection of such diseases.
Types of Immunological Products
VACCINES
Vaccine is a biological preparation that consists of
either a whole organism or a part of it against which
immunization has to be achieved.
Vaccines provide active immunity as they stimulate
the immune system of the recipient to produce T
cells or antibodies that impede the attachment of
infectious agents and promote their destruction.
Types of Vaccines:
Live Vaccines:
 These are preparations of live bacteria,
viruses or other agents which, when
administered by an appropriate route, cause
subclinical or mild infections. In the course
of such an infection the components of the
microorganisms in the vaccine evoke an
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immune response which provides protection
against the more serious natural disease.
 Examples: Vaccinia (Smallpox), BCG (TB)
Killed Vaccines:
 Killed vaccines are suspensions of bacteria,
viruses or other pathogenic agents that have
been killed by heat or by disinfectants such
as phenol, ethanol or formaldehyde.
 Killed organisms cannot replicate and cause
an infection. Thus every dose of killed
vaccine must have an antigenic material to
increase the immunogenic response
 Since all the components of the micro-
organism are present, it may be toxic to the
body. Thus it is recommended to divide the
vaccine into booster doses which may be
given at regular intervals of time.
 Examples: Polio, Typhoid, Pertussis,
Cholera, Plague, Rabies.
Toxoid Vaccines:
 Toxoid vaccines are preparations derived
from the toxins that are secreted by certain
species of bacteria.
 In the manufacture of such vaccines, the
toxin is separated and treated chemically
(Formaldehyde) to eliminate toxicity but not
immunogenicity.
 This process is called as toxoiding and the
end product is termed as Toxoid or Formol
toxoids.
 Examples: Tetanus, Diphtheria, Botulism,
Clostridial infections of farm animals.
Cell components or Subunit Vaccines
 Instead of using whole cells which may
consist of undesirable reactogenic
components, vaccines are prepared from
purified protective components.
 Such vaccines is that they evoke an immune
response only to the component, or
components, in the vaccine and thus induce
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a response that is more specific and
effective.
 Examples: Hemophilus influenzae type b,
Neisseria meningitidis ACWY, Hepatitis b
etc.
Conjugate Vaccines
 Some antigens which are used to prepare
vaccines are less immunogenic and do not
give appropriate responses.
 Such antigens are conjugated to certain
immunogenic carriers which improve the
immunogenic response.
 Example: Glyco- Conjugate Vaccine of
Neisseria meningitidis with carrier protein is
CRM197
Adjuvants
 Heterogeneous collection of substances
which enhance the immune response.
 Examples: Aluminium hydroxide gel
(hydrated aluminium oxide) and aluminium
phosphate are the only ones in general use in
human vaccines.
 A much wider range of substances including
oily emulsions, saponin, immune-
stimulating complexes (ISCOMS),
monophosphoryl lipid A and others are used
in veterinary vaccines and some are under
investigation for use in human vaccines
QUALITY CONTROL
Mainly to provide assurances of both the probable
efficacy and safety of every batch of every product.
There are two ways
1. In-process control
2. Final product control
a. Assays
b. Safety tests
In-process Control
 In-process quality control is the control
exercised over starting materials and
intermediates.
 The toxoid concentrates used in the
preparation of the vaccines have been much
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diluted and, as the volume of vaccine that
can be inoculated into the test animals
(guinea-pigs) is limited, the tests are
relatively insensitive. In-process control,
however, provides for tests on the undiluted
concentrates and thus increases the
sensitivity of the method at least 100-fold.
Final Product Control
Assays:
 Vaccines containing killed microorganisms
or their products are generally tested for
potency in assays in which the amount of the
vaccine that is require to protect animals
from a defined challenge dose of the
appropriate pathogen, or its product, is
compared with the amount of a standard
vaccine that is required to provide the same
protection.
 The number of survivors in each group is
used to calculate the potency of the test
vaccine relative to the potency of the
standard vaccine by the statistical method.
 The potency of the test vaccine may be
expressed as a percentage of the potency of
the standard vaccine
 Vaccines containing live microorganisms
are generally tested for potency by
determining their content of viable particles.
 Example: In the case of BCG vaccine,
dilutions of vaccine are prepared in a
medium which inhibits clumping of cells,
and fixed volumes are dropped on to solid
media capable of supporting mycobacterial
growth. After a fortnight the colonies
generated by the drops are counted and the
live count of the undiluted vaccine is
calculated.
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Safety Tests
 Bacterial vaccines are regulated by
relatively simple safety tests. Those vaccines
composed of killed bacteria or bacterial
products must be shown to be completely
free from the living microorganisms used in
the production process.
 Those vaccines prepared from toxins, for
example, diphtheria and tetanus toxoids,
require in addition, a test system capable of
revealing inadequately detoxified toxins.
 This can be done by inoculation of guinea-
pigs, which are exquisitely sensitive to both
diphtheria and tetanus toxins.
 A test for sensitization of mice to the lethal
effects of histamine is used to detect active
pertussis toxin in pertussis vaccines.
 With killed vaccines the potential hazards
Parametric Release
are those due to incomplete virus
inactivation and the consequent presence of
residual live virus in the preparation.
 With attenuated viral vaccines the potential
hazards are those associated with reversion
of the virus during production to a degree of
virulence capable of causing disease in
recipients
Tests of general Applications
Bioburden
 A successful sterilization process is
dependent on a product having a low pre-
sterilization bioburden. Sterilization should
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be considered as the removal of the
bioburden.
 This will also be true of the individual
ingredients, which must have low levels of
microbial contamination or else there is a
danger that the contaminants will find their
way into the final product or be a source of
pyrogens.
 The bioburden is an estimate of the total
viable count of microorganisms present pre-
sterilization, and a knowledge of the
resistance characteristics of these organisms
is often an integral part of the sterility
assurance calculation.
 Sterilization process should be chosen in
such a way that all micro-organisms are
highly resistant.
 As there are significant limitations with the
test for sterility, many authorities place
considerable reliance on the validation and
reliable performance of sterilizers and their
sterilization cycles.
 Parametric release takes this reliance a
step further by allowing batches of
terminally sterilized products to be released
without being subjected to the test for
sterility.
 Validation studies would include heat
distribution, heat penetration, bioburden,
container closure and cycle lethality studies.
 For a product to be subject to parametric
release, pre-sterilization bioburden testing
on each batch would be completed, and the
comparative resistance of isolated spore-
formers checked.
 In practice this requires confirmation that
each part of the manufacturing process has
been satisfactorily completed, the initial pre-
sterilization bioburden is within agreed
limits, that the controls for the sterilizing
cycle were satisfactory and that the correct
time cycles were achieved.
 Clearly reproducibility, regular monitoring
and documentation are required
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Post Marketing Surveillance of
Vaccines
Vaccine Adverse Event Reporting System
 Co-administered by FDA and CDC
 Reporting by paper or electronic versions of
a standard form
o Serious AE reports are manually
reviewed by medical officers to
detect unexpected events
o Nonserious reports assessed
primarily through data mining
Post-licensure Rapid Immunization Safety
Monitoring System (PRISM)
• Integral part of Mini-Sentinel dedicated to
vaccine safety
• 42 million individuals
o 3 national health plans, 8 vaccine
registries
• Evidence reviews for key health outcomes
underway
• Developing methods
o Identify signals without pre-
specifying outcomes
o Perform sequential regression using
propensity scores
Stability of Vaccines
1. Choice of stability indicating parameters and
frequency of testing
2. Cumulative age of an antigen in the final
product
3. Stability of a final lot
a) Vaccine formulation
b) Vaccine presentation, container and
closure system
c) Stability of freeze-dried vaccines
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Choice of stability indicating parameters and
frequency of testing
Depending on the nature of the antigen and other
components as well as on the manufacturing
process, stability indicating parameters should be
selected on a case-by-case basis. In the selection of
stability indicating parameters, the potential clinical
implications of the observed changes must always
be considered. Ideally, stability indicating
parameters should reflect the link between vaccine
quality and efficacy or safety as demonstrated in
clinical trials.
For most vaccines, potency is considered as a
stability indicating parameter that reflects potential
impact of environmental conditions on the
immunogenicity and subsequent protective efficacy
of a vaccine.
Cumulative age of an antigen in the final product
The stability of the characteristics of a final product
should be guaranteed during the whole shelf-life,
irrespective of the age of the intermediates at the
time they are used in the production process. Total
age of all components at the end of shelf-life is
considered as cumulative age of the product. In
practice, stability data of the final product should
include the data generated on the intermediates of
different ages used in the final formulation
Stability of a final lot
a) Vaccine formulation:
The stability of a final lot of vaccine depends of the
stability of all intermediates as well as of the final
formulation. Therefore, data on stability of the
intermediates as well as stability data of the final
formulation should be submitted to the National
Regulatory Authority.
In the case of combined vaccines, the stability of
each component should be assessed and data
included in the manufacturers dossier.
b) Vaccine presentation, container and
closure system
In addition to the data on stability of the final
formulation, other factors that may affect vaccine
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stability during its use should also be tested in the

stability study.

Potential interactions between the vaccine and

container and closure system are particularly
important for vaccines in liquid form.

c) Stability of freeze-dried vaccines

Data to support proposed use of vaccine after
reconstitution, maximum storage period, and
storage conditions should be generated as part of the
stability study performed on the final lot. In the
assessment of freeze-dried vaccines, residual
moisture should be specified.

Reconstitution period (time needed for

reconstitution and appearance of reconstituted
vaccine should be defined.

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MISCELLANEOUS
DETERMINATIONS
AND TESTS

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DETERMINATION OF Karl Fischer Titration:

 Classical method of titration
WEIGHT/mL  Invented in 1935

Karl Fischer Reagent:

The weight per milliliter is determined by dividing
the weight in air, expressed in g, of the quantity of
liquid that fills a pycnometer at the specified
temperature by the capacity, expressed in ml, of
the pycnometer at the same temperature.
The capacity of the pycnometer is ascertained from
the weight in air, expressed in g, of the quantity of
water required to fill the pycnometer at that
temperature.
A solution of iodine and sulphur dioxide in a
mixture of pyridine and methanol.
Principle
The titrimetric determination of water is based upon
the quantitative reaction of water with an anhydrous
solution of sulphur dioxide and in the presence of a
buffer that reacts with hydrogen ions.
2H₂O+ SO₂+I₂→2HI+H₂SO₄

The weight of a litre of water at specified Characteristics of the Karl Fischer Titration
temperatures when weighed against brass weights in Method:
air of density 0.0012 g per ml is given in the 1. Moisture content can be determined accurately
2. Measurements can be taken over short periods
following table. of time
3. Measurements can be taken using small
Ordinary deviations in the density of air from the samples
above value, here taken as the mean, do not affect 4. The method can be used with liquid, solid and
the result of a determination in the significant gaseous sample
figures prescribed for Pharmacopoeial substances. 5. The method is suitable for unstable substances
that can alter when heated
Temperature °C Weight of a litre of water
Working:
20 997.18
25 996.02
Water and iodine are consumed in a 1:1 ratio in the
above reaction.
30 994.62
Once all of the water present is consumed, the
presence of excess iodine is detected Volta
Water Content metrically by the titrator‘s indicator electrode.
That signals the end point of the titration. The
Determination amount of the water present in the sample is
calculated based on the concentration of iodine in
There are following methods: the Karl Fischer titrating reagent (i.e., titer) &
amount of Karl Fischer reagent consumed in the
1. Method 1 (Titrimetric) titration.
2. Method 2 (Azeotropic)
3. Method 3 (Gravimetric) Apparatus:
 A closed system consisting one or two
Method 1 (titrimetric) automatic burets
1) Karl Fischer
 A tightly covered titration vessel fitted with
2) Residual
electrodes
3) Coulometric
 A magnetic stirrer
 A suitable desiccant

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Standardization:
 To the titration vessel, add methanol R, dried
if necessary, or the solvent recommended by
the supplier of the titrant.
 Where applicable for the apparatus used,
eliminate residual water from the
measurement cell or carry out a pre-
titration.
 Introduce a suitable amount of water in an
appropriate form and carry out the titration,
stirring for the necessary time.
 The water equivalent is not less than 80% of
that indicated by the supplier.
 Standardize the titrant before the first use
and at suitable intervals thereafter.
Procedure:
 Unless otherwise specified, transfer 35 to
40 ml of methanol or other suitable solvent
to the titration vessel, and titrate with the
Reagent to the electrometric or visual
endpoint to consume any moisture that may
be present.
 Quickly add the test preparation, mix, and
again titrate with the reagent to the
electrometric or visual endpoint.
 Calculate the water content of the specimen,
in mg, taken by the formula (SF)
 S is the volume, in ml, of the reagent
consumed in the second titration
 F is the water equivalence of the reagent.
Visual Detection:
In case of colorless solution that is titrated directly,
the end point may be observed visually as a change
in color from canary yellow to amber.
Electrical Detection:
When alkyl sulphate moiety is developed then
ammeter show deflection. When ammeter show
persistent deflection for 30 sec then it is assumed
that end point is reached.
Residual Titration
Principle:
In the residual titration, excess reagent is added to
the test specimen, sufficient time is allowed for the
Adnan Sarwar Chaudhary
reaction to reach completion and the unconsumed
reagent is titrated with a standard solution of water
in a solvent such as methanol.
The residual titration procedure avoids the
difficulties that may be encountered in the direct
titration of substances from which the bound water
is released slowly.
Apparatus, Reagents and test preparation:
Use same as above
Procedure:
Were the individual monograph specifies that the
water content is to be determined by method 1b,
transfer 35-40 ml of methanol or other suitable
solvent to the titration vessel, and titrate with the
reagent to the electrometric or visual endpoint.
Quickly add the test preparation, mix , and add an
accurately measured excess of the reagent.
Allow sufficient time for the reaction to reach
completion, and titrate the unconsumed reagent
with standardized water solution to the
electrometric or visual endpoint.
Calculate the water content of the specimen, in mg,
taken by the formula:
F(X’-XR)
F is the water equivalence factor for the reagent
X’ is the volume, in ml, of the reagent added after
introduction of the specimen
X is the volume, in ml, of the standardized water
solution required to neutralized the unconsumed
reagent
R is the ratio, determined by the standardization of
the water solution for residual titration (ml of the
reagent/ ml of water solution)
Coulometric Titration
Principle:
 The coulometric titration of water is based
upon the quantitative reaction of water with
sulphur dioxide and iodine in an anhydrous
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medium in the presence of a base with
sufficient buffering capacity.
 In contrast to the volumetric method,
iodine is produced electrochemically in the
reaction cell by oxidation of iodide.
 The iodine produced at the anode reacts
immediately with water and the sulphur
dioxide contained in the reaction cell.
 The amount of water in the substance is
directly proportional to the quantity of
electricity up until the titration endpoint.
 When all of the water in the cell has been
consumed, the endpoint is reached and thus
an excess of iodine appears. 1mole of iodine
corresponds to 1mole of water, a quantity of
the electricity of 10.71C corresponds to 1mg
of water.
 Moisture is eliminated from the system by
pre-electrolysis. Individual determinations
can be carried out successively in the same
reagent solution, under the following
conditions:
o Each component of the test mixture
is compatible with the other
components
o No other reactions take place
o The volume and the water capacity
of the electrolyte reagent are
sufficient
Apparatus:
 Absolutely tight system fitted with the
necessary electrodes and a magnetic stirrer
 The reaction cell consists of a large anode
compartment and a smaller cathode
compartment separated by diaphragm
Adnan Sarwar Chaudhary
Test Preparation:
 Where the specimen is a soluble solid,
dissolve an appropriate quantity,
accurately weighed, in anhydrous
methanol or other suitable solvents.
 Liquids may be used as such or as accurately
prepared solutions in appropriate anhydrous
solvents.
 Where specimen is an insoluble solid the
water may be extracted using a suitable
anhydrous solvent from which an
appropriate quantity, accurately weighed,
may be injected into anolyte solution.
 Alternatively an evaporation technique may
be used in which water is released &
evaporated by heating the specimen in a
tube in a stream of dry inert gas, this gas
being then passed into the cell.
Procedure:
 Using a dry syringe, quickly inject the test
preparation accurately measured and
estimated to contain
 0.5 to 5 mg of water, or as recommended by
the instrument manufacture into the anolyte,
mix, and perform the coulometric titration to
the electrometric end point.
 Read the water content of the test
preparation directly from instrument‘s
display, and calculate the % that is present in
the substance.
 Perform a blank determination, and make
any necessary corrections.
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Method 2 (azeotropic-toulene
distillation)
Apparatus:
Use a 500 ml glass flask A connected by means of a
trap B to a reflux condenser C by ground glass
joints.
Procedure:
 Place the accurately weighed quantity of the
substance in the flash to the nearest
centigram.
 Place about 200ml of toluene in the flask.
 Connect the apparatus.
 Fill the receiving tube E with toluene poured
through top of condenser.
 Heat the flask gently for 15 minutes until the
toluene begins to boil.
 Distill out the water at the rate 2 drops per
second.
 Continue distillation for 5 minutes.
 Remove the heat. Allow the flask to cool to
room temp
 Scrub any adhered water droplets to the
walls of the tube, with the brush.
 When the water and toluene have separated
completely, read volume of water and
calculate the %age that was present in the
substance.
METHOD 3 (GRAVIMETRIC)
Procedure for Chemicals
Proceed as directed in the individual monograph
preparing the chemicals as directed under Loss On
Drying.
Adnan Sarwar Chaudhary
Procedure for Biologics
Proceed as directed in the individual monograph
LOSS ON DRYING
Definition:
The loss on drying test is designed to measure the
amount of water and volatile matters in a sample
when sample is dried under specified condition.
Procedure USP:
 Mix and weigh the substance.
 Unless otherwise directed in the monograph.
 Reduce size about 2mm if in crystal form.
 Take a covered petri-dish dry it for about 30
minutes under same conditions.
 Weigh it without cover.
 Put the sample in it.
 Weigh content with petri-dish.
 Spread the sample not more than 10mm in
depth.
 Place it in oven under conditions given in
monograph.
 Upon opening the oven promptly cover the
dish, allow to come in room temperature in
dessicator.
 If sample melt at specified temperature then
kept sample for one to two hours at
temperature 5-10℃ below its melting point.
 In case of capsules and tablets use content of
not less than 4 capsules.
 If drying in vacuum over a dessicator is
directed suitable vacuum drying apparatus
should use.
 If drying in a capillary –stoppered bottle in
vacuum is directed, use bottle with stopper
having
 225±25 diameter and maintain pressure of
5mm or less of mercury.
 Formula:
 LOD is calculated by:-
IDENTIFICATION TESTS
Definition:
It is performed to measure;
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 Severity and Number of subjects: 6

 Potential of health risk due to drug problems
Weight range: 1.8 – 2.2 kg
Purpose:
Requirement: These rabbits are placed under
The purpose of identification test is to check drug:
observation and food is not given for 18-24 hrs
 Bio-analysis e.g., mechanism of action before experiment
 Bioavailability
Procedure:
 Interaction of one drug with other
 Inject subcutaneously, a quantity of insulin
Methods: injection, which should cause convulsion in
atleast of the animals
These are perfomed both;  Immediately after convulsions, inject 5ml of
50% dextrose solution intravenously…
A. In vivo
convulsions should be stopped in atleast 4 of
B. In vitro
the animals and they should remain alive for
atleast 3 days after
IN VIVO
Specification:
IDENTIFICATION TESTS: This is specified in the monograph

In vivo identification tests discussed are for: Test For Ophthalmic Ointment
1. Atropine And Ophthalmic Solution
2. Insulin Injection (Isophlurophate):
3. Ophthalmic ointments and Ophthalmic Requirement:
solutions
Subject: Rabbit
1. Test for Atropine:
Number of subjects: 3
General characteristics of atropine are:
Procedure
1. Belladona Alkaloid
 Take 100mg of isoflurophate ointment or
2. Affinity for muscarinic receptors
0.1ml of solution in the right eye of each of
3. Cause mydriasis in eye by blocking
three rabbits
cholinergic activity
 Observe the constriction of pupil
Procedure for test:
RESULT:
 First take sufficient number of tablets or
If atleast 20mm decrease occurs then it is confirmed
injection which contain 0.6 to 1 mg Atropine
sulphate
 Dissolve it in 10 ml water IDENTIFICATION TESTS
 Take 1 ml from this and instill in rabbits eye
and observe IN VITRO
Result: For some drugs in vitro tests are performed and
If dilation occurs within 2 hrs atopine is confirmed compared with standard

Test For Insulin Injection:  The IR absorption spectra for chlorthiazide

dispersed in mineral oil should show
Subject: Rabbit

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maxima equal to the chlorthiazide refrence
standard.
 When transmitted light is passed through
Quabin dissolved in sulphuric acid produces
dark red color and produce greenish
fluorescence in reflected light..
 Blue fluorescence is exhibited in case of
solution of quinine gluconate in dilute
sulphuric acid, on addition of few drops of
HCl fluorescence disappears.
TOXICITY TESTS
Toxicology:
It is the study of the adverse effects of the physical,
chemical or biologic agents on living organisms and
the ecosystem.
Tests for Toxicity
1. Single Dose Acute Toxicity Test
Toxicity produced by a pharmaceutical when it is
administered in one or more doses during a period
not exceeding 24 hrs.
Purpose & Procedure:
Test is performed to identify doses causing no and
maximum toxic effects.
For acute toxicity tests two routes are used:
i. The route intended for human administration &
ii. Intravenous administration if feasible
Observation
Animals are kept under observation for 14 days
after administration of pharmaceutical and
following observations are noted and recorded
 Mortalities
 Time of onset
 Duration
 Reversibility of toxicity
If primary data supports single dose safety/kinetic
studies in humans, the toxicity studies should be
designed to access dose response relationship and
pharmacokinetics.
Adnan Sarwar Chaudhary
2. Repeated Dose Toxicity
Purpose:
 Development of safe medicinal products that
need repeated administration to patients.
 To characterize toxicological profile of test
compound following repeated
administration.
 To identify potential target organs of
toxicity & potential reversibility of toxic
effects.
General Recommendations Concerning The
Experimental Animal:
Species: it should be chosen based on similarity to
humans with regard to pharmacokinetic profile and
biotransformation.
 Sexes: Equal no. of male and female animals
should be taken.
 Size: It should be sufficient to allow
meaningful scientific interpretation of data
generated.
 No. of Species: It should be carried out in
two species of mammals, one of which,
must be non- rodent
 Animal Husbandry: A high standard of
animal husbandry is required &
Environmental conditions should be
controlled.
General Recommendations Concerning Dose and
Administration:
Duration of Administration: it depends on the
duration of proposed therapeutic use in humans.
 Route of Administration: same route should
be chosen as intended for humans. Other
routes are selected if justified on the basis of
pharmacology.
 Frequency of Administration: it should be
taken on the basis of intended clinical
dosing regimen and
toxicological/pharmacokinetic/pharmacodyn
amics profile of test compound.
 Dose levels: Treatment should include:
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o A low dose: sufficient to produce a  In it we determine drug‘s:

pharmacodynamic effect or the o Pharmacokinetics
desired therapeutic effect. o Metabolism &
o A high dose: selected to enable o Mechanism of action
identification of toxicity for target
organ. Phase 2
o Intermediate dose: geometric mean  Determine the short term effects of drugs &
between high and low dose.  Evaluate effectiveness of treatment

Observation: Phase 3
 Pre-treatment and control values; historical  It is conducted with several thousand
control data should be available for patients to gather information about
morphological, biochemical and physiologic effectiveness and safety.
variables studied, both for rodents and non-  It is done prior to human clinical
rodents. For non-rodents pre-treatment investigations
values should be obtained from animal used
Advantages:
in the study.
 Any type of toxicity test can be evaluated
 Monitoring during the study; followings are
 Mechanism of toxicity test can be evaluated
monitored: Food intake, General behavior,
Body weight, Heamatological parameters, Why Required:
Clinical chemistry, Urinalysis and Toxicity tests are required for different
ophthalmology.
 Drugs
Data Analysis, Presentation Of Results And  Toxoids &
Conclusions:  Containers
 Study report should reflect all the raw data
and information gathered during the study. Example:
 Group summary values should be presented. Diphtheria toxoid
 Finally, a conclusion based on study results
Procedure:
should be drawn.
For toxicity test of diphtheria toxoid
Methods for Toxicity Testing:
 Take four healthy pigs of weight 300-400g
1. Clinical investigation; chemicals are
 Inject 2ml diphtheria toxoid subcutaneously
administered to human subjects with careful
observation and lab measurements. Result
2. Epidemiological studies; observation of No symptoms of diphtheria toxoid should appear
humans that have been exposed to within 30 days
xenobiotics in normal course of their life
and occupation. EVALUATION OF
3. Reports of drug adverse reactions; Reports
are submitted to FDA after drug has been
approved.
OINTMENT
Phases of Clinical Investigation: Ointment:
The word ointment derived from Latin word
Phase 1 ―UNGUENT‖ means anoint with oil.
 Drug is tested in a small group of 20-80
patients
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Anoint with oil means rub with oil, or covered with effects on the skin or carry medicaments for
oil. treating certain topical ailments.
 Any greasy or oily semi solid preparation,
Ointments are semisolids intended for external
usually medicated, that can be applied
application to the skin or mucous membranes
externally to the skin in order to heal, soothe
OR or protect it.

Ointments are semisolids intended for external Types of Ointment bases:

application to the skin or mucous membranes that There are five (5) classes or types of ointment bases
usually contain less than 20% water and volatiles which are differentiated on the basis of their
and more than 50% hydrocarbons, waxes, or physical composition.
polyols as the vehicle and provide emollient,
These are:
protectants, or lubricants properties.
1. Oleaginous bases/hydrocarbon bases
 Ointments are semisolid preparations 2. Absorption bases
intended for topical application. They are 3. Water in oil emulsion ointment bases
used to provide protective and emollient 4. Oil in water emulsion ointment bases
5. Water soluble or water miscible bases
Oleaginous Absorption Water/Oil Oil/Water Water
Ointment Bases Ointment Bases Emulsion Emulsion miscible
Ointment Bases Ointment Bases Ointment
Bases
Composition
oleaginous oleaginous oleaginous oleaginous Polyethylene
compound base + base + base + Glycols
s w/o surfactant water (< 45% water (> 45% (PEGs)
w/w) w/w) + o/w
+ w/o surfactant
surfactant (HLB >9)
(HLB <8)
Water
Content anhydrous anhydrous Hydrous hydrous anhydrous,
hydrous
Affinity for
Water hydrophob hydrophilic Hydrophilic hydrophilic hydrophilic
ic
Spreadability
difficult difficult moderate to easy moderate to
easy easy
Washability
Non Non washable non- or washable washable
washable poorly
washable
Stability
oils poor; oils poor; unstable, unstable, stable
hydrocarbo hydrocarbons especially especially
ns better better alkali soaps alkali
and soaps and
natural natural

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colloids colloids; non

ionics better
Drug
Incorporatio solids or solids, oils, solids, oils, solid and solid and
n Potential oils (oil and and aqueous aqueous
solubles aqueous aqueous solutions solutions
only) solutions solutions (small
(small (small amounts)
amounts) amounts)
Drug Release
Potential* poor poor, but > fair to good fair to good good
oleaginous
Occlusiveness
yes yes Sometimes no no
Uses
protectants protectants, emollients, emollients, drug vehicles
, emollients cleansing vehicles for
emollients (+/-), creams, solid,
(+/-), vehicles for vehicles for liquid, or non
vehicles aqueous solid, hydrolyzable
for solutions, liquid, or non drugs
solids, and hydrolyzable
non drugs
hydrolyzable
drugs

Color
Quality Control Tests: Colour should be colourless to yellow.

1. Appearance and colour USP Weight variation tests or minimum

2. Weight variation tests BP or minimum fill
3. Particle size determination BP
4. Microbial content test USP
5. Metal contents in ophthalmic ointments BP
6. Sterility tests
7. Potency or content uniformity test BP
8. Viscosity
9. Leakage tests
10. Homogenicity
Appearance and colour:
Procedure
Transfer the ointment to a suitable test tube and examine
the sample in front of light source.
Acceptance rejection criteria:
Translucent sample is accepted.
fill
 Test is applied only to those containers that
contain not more than 150 g or mL of preparation.
 Select 10 filled containers, remove label and
weight them individually
 Remove the contents and weigh the empty
containers individually.
 Take difference of full and empty container for
getting contents.
 Take average of 10 tablets and its weight should
not be less than labelled amount.
Acceptance/rejection criteria:
In 2 stages, in S1 select 6 units if not meet criteria then
on S2 select 20 more units and total units is 30.

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Sample size S1= for 10 units S2=for 30  These microbes effect the skin and the
For ≤60g Net wt of Net wt of ointment also used for the skin condition is
contents of any contents of not already compromised.
single unit more than 3
should not be units should be
less than 90% of less than 90% of Metal particle test:
the labelled
labelled amount. amount.  This test is required only for ophthalmic
For 60-150g Net wt of Net wt of ointments. It is performed using 10 ointment
contents of any contents of not
single unit more than 1 tubes.
should not less unit should be  The content from each tube is completely
than 95% of the less than 95% of removed onto a clean 60 - mm - diameter
labelled amount. labelled
petri dish which possesses a fl at bottom
amount
 The lid is closed and the product is heated at
Particle size determination: 85 ° C for 2 h. Once the product is melted
 Dilute a specific quantity of sample with and distributed uniformly, it is cooled to
equal volume of glycerol or liquid paraffin room temperature. The lid is removed after
or specified solidification.
in monograph.  The bottom surface is then viewed through
 Mount the diluted sample on slide and an optical microscope at 30 magnification.
examine random fields microscopically The viewing surface is illuminated using an
using microscope external light source positioned at 45° on the
providing adequate resolution for top.
observation of small particles.  The entire bottom surface of the ointment is
 Count the number of particles with diameter examined, and the number of particles are
above or below than that specified in 50µm or above counted using a calibrated
monograph. eyepiece micrometer.
 Compare the percentage with official limits.
Acceptance rejection criteria:
Acceptance rejection criteria  The number of such particles in 10 tubes
Diameter ≥10µm ≥ 25µm ≥ 50µm should not exceed 50, with not more than 8
of particles particles in any individual tube.
No. Of Not more Not more Not more
particles than 12/ml than 5/ml than 2/ml  If these limits are not met, the test is
repeated with an additional 20 tubes.
Microbial content test:  In this case, the total number of particles in
30 tubes should not exceed 150, and not
 Except ophthalmic preparations, tropical more than 3 tubes are allowed to contain
applications are not required to be sterile. more than 8 particles.
 They must meet acceptable standards for
microbial content. Sterility test:
 Microbial limits are stated for certain
articles in USP.  Applied to the products that required to be
sterile such as ophthalmic preparations.
Example:  Ophthalmic semisolids should be free from
 Betamethasone valerate ointment, must clear anaerobic and aerobic bacteria and fungi.
the test for absence of Staphyloccus aureous
and Pseudomonas aeruginosa. Methods:
 Direct inoculation method
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 Membrane filtration method
i. Membrane filtration method:
 In the membrane filtration method, a
solution of test product (1%) is prepared in
isopropyl myristate and allowed to penetrate
through cellulose nitrate filter with pore size
less than0.45µm.
 If necessary, gradual suction or pressure is
applied to aid filtration.
 The membrane is then washed three times
with 100 - mL quantities of sterile diluting
and rinsing fluid and transferred aseptically
into fluid thioglycolate (FTG) and soybean –
casein digest (SBCD) medium.
 The membrane is finally incubated for 14
days.
 Growth on FTG medium indicates the
presence of anaerobic and aerobic bacteria,
and SBCD medium indicates fungi and
aerobic bacteria.
 Absence of any growth in both these media
establishes the sterility of the product.
ii. Direct inoculation method:
 In the direct - inoculation technique, 1 part
of the product is diluted with 10 parts of
sterile diluting and rinsing fluid with the
help of an emulsifying agent.
 Incubated in FTG and SBCD media for 14
days.
Criteria:
In both techniques, the number of test articles is
based on the batch size of the product. If the batch
size is less than 200 the containers, either 5% of the
containers or 2 containers (whichever is greater) are
used. If the batch size is more than 200, 10
containers are used for sterility testing.
Acceptance rejection criteria:
 If no evidence of microbial growth, sample
is sterile.
 If evidence of microbial growth, sample is
non sterile.
Adnan Sarwar Chaudhary
Potency/content uniformity
test:
 In this test the different analytical techniques
are used.
 Assay the 10 units individually, if test failed
20 more containers are tested.
 Drug assay is performed by using different
analytical techniques e.g. titrimetric assay.
 Conduct the assay on the amount of the
material that drains from the individual
container.
 Adjust the degree of the dilution so that the
concentration of the active ingredient in the
final solution is of same order as that
obtained in the assay procedure.
Acceptance rejection criteria:
For 10 doses unit acceptance criteria:
 Not more than 1 individual content, is
outside the limit of 85%-115% of average
labelled amount.
 Not a single unit should be outside the limit
of 75%-125% of average labelled amount.
Rejection criteria:
 More than 3 individual content is outside the
limit of 85%-115% of average labelled
amount or
 One or more is outside the limit of 75%-
125% of average labelled amount.
If this step reject then take 20 more units and tested
individually.
For 30 unit acceptance criteria:
 Not more than 3 units are outside the 85%-
115% of average labelled amount
 No one is outside the 75%-125% of average
content.
Viscosity:
 Viscosity is a property of liquids that is
closely related to resistance to flow.
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 Force required to moves one plane surface
continuously past another under specified
conditions.
 Basic unit is poise=100 centipoises.
 Viscosity of the water as a reference
material and all viscosities compare with
this standard.
 Specifying of temperature is more important
because viscosity decreases as temperature
raised.
 Viscosity take at 20 degree centigrade, and
for water is 1.
Measurement of viscosity:
Many capillary tube viscometers are used.
 Ostwald viscometers
 Ubbelohde viscometers
Adnan Sarwar Chaudhary
Leakage test
 This test is mandatory for ophthalmic
ointments, which evaluates the intactness of
the ointment tube and its seal.
 Ten sealed containers are selected, and their
exterior surfaces are cleaned.
 They are horizontally placed over absorbent
blotting paper and maintained at 60± 3 ° C
for 8 h.
Acceptance rejection criteria:
 The test passes if leakage is not observed
from any tube.
 If leakage is observed, the test is repeated
with an additional 20 tubes.
 The test passes if not more than 1 tube
shows leakage out of 30 tubes.
Homogenicity:
Content of ointment is placed on glass slab and
spread in form of thin layer. And then observe
under light source.
Acceptance criteria:
Ointment is homogenous if there are no granules
present.
ASH CONTENT
DETERMINATION:
Definition:
Ash content is measure of total amount of minerals
present in a product.
Mineral Content:
It is the measure of total amount of specific
inorganic compounds present in a product e.g Ca,
Na, Cl, Mg, Cu, Mn, Zn etx
Determination Methods BP:
There are two methods:
1. Acid insoluble ash value
2. Determination of sulphated ash
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Acid insoluble ash value:
Method 1:
 Ash content is boiled with 25ml Hcl (1:2:5)
for 5 min in water bath,covering the dish
with watch glass.
 It is then filtered through ashless filter paper
No.40.
 The residue is washed with water until free
of acid.
 It is then ignited at 600˚C for 20min.
 It is then cooled and weighed.
Method 2:
 Ash insoluble in Hydrochloric acids the
residue obtained after extracting the Sulfated
or Total ash with Hydrochloric acid,
calculated with reference to 100g of drug.
 To the crucible containing the residue from
the determination of Sulfated or Total ash,
add 15ml of water and 10ml of Hydrochloric
acid, cover with a watch glass, boil the
mixture gently for 10min and allow to cool.
 Filter through an ashless filter.
 Wash the residue with hot water until the
filtrate is neutral, dry, ignite to dull redness,
allow to cool it in a dessicator and weigh.
 Repeat until the difference between 2
consecutive weighing is not more than 1mg.
2. Sulphated –Ash value:
Method 1:
 Heat platinum dish to redness for 10min.
 Allow to cool in a dessicator and weigh.
 Unless otherwise specified in the
monograph, place 1g of substance in dish
moisten with Sulphuric acid.
 Ignite gently.
 Again moisten with Sulphuric acid, again
ignite at 800˚c.
 Cool, weigh, ignite for 15min and repeat this
procedure until two successive weighing do
not differ by more than 0.5mg.
Adnan Sarwar Chaudhary
Method 2:
 Ignite suitable crucible at 600± 50℃ for
30min.
 Cool in a dessicator.
 Place sample in crucible n weigh.
 Moisten with sulphuric acid usually (1ml).
 Heat gently at low temperature.
 Cool, again moisten, heat until white fumes
no longer evolved.
 Then at 600±50℃ until residue completely
incinerated.
 Cool in a dessicator, weigh and calculate
%age of residue.
 If amount of residue exceed the prescribed
limit, repeat the procedure.
ALKALINITY OF GLASS
―It is measure of ability of solution to neutralize
acids to equivalence point of carbonates &
bicarbonates‖.
 Alkalinity is basically/mainly due to
carbonates due to its common presence in
atmosphere.
 It may also be due to borates, hydroxides,
silicates & sulfides, etc.
 Unit of alkalinity is mEq/L.
 Commercially, ppm is used.
How Alkalinity of Glass Occurs?
 Glass contains sodium & potassium oxides
which are hygroscopic & absorbs the
moisture from air. This causes the oxides to
get converted to sodium & potassium
carbonates. These are also hygroscopic.
 In sea water, Na & K carbonates in unstable
glass may leach out leaving fragile, cracked,
flaked glass with frosty appearance.
Treatment of Unstable Glass:
They can be treated in different ways but the one is
as follows:
 Wash the glass in running tap water.
 Soak in distilled water.
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 Dry in 2 baths of alcohol (This prevents 3. Chemical Resistance of Test

disintegration, breakdown of glass & Equipments used in this test should be of high
improves appearance). quality & area should be from fumes & dust.
 To impede (delay) disintegration, apply
Apparatus:
organic lacquer.
 Auto clave (Temp. 121 ± 0.5)
 Store in environment of humidity not higher
 Mortar & pestle
than 40%.
 8 inch sieves (20, 40)
Types of Glass:
Type General Type of Test
Reagents
Description  Special distilled water
I Highly resistant, Powdered glass  Methyl red solution
Borosilicate test
glass 4. Powdered Glass Test:
II Treated Soda- Water attack test  This test is official in USP & IP.
lime glass
III Soda-lime glass Powdered glass  In this test, alkalinity as well as other glass
test constituents are measured.
IV or O General purpose Powdered glass
soda-lime test Procedure:
These are classified by Pharmacopoeial  Rinse with water (6 or more containers)
preparations according to the resistance to chemical  Dry in air
attack. Types I, II & III are for parenteral use While  Crush (25mm in size)
types O is for non-parenteral use.  Divide in 3 portions & place 1 in the mortar.
 Empty mortar in sieve no. 20 & repeat the
Type of Glass Test same procedure for the remaining two
portions.
1. Crushed Glass Test:
This test is official in USP.  Then pass through the sieve no. 40 & empty
in the mechanical shaker for 5 minutes.
 Container may be crushed & sieved to  Spread sample on glazed paper & pass the
produce definite particles. magnet over the material to eradicate the
 The control of particle size & weight of iron particles of sieves if present.
powder ensures constant surface area  Then transfer it in conical flask & wash.
exposed to solution.  Dry for 20 minutes at 140 degrees.
 All of glass is examined & extraction is  Use test solution within 48 hours.
enhanced by rough surface of particles.
 This test can be used to determine nature of 5. Water Attack Test:
glass.  This test is used for determining alkali
leaching out from surface of container.
2. Whole Container Test:  Mainly based on the alkali released from
 This test is official in European, British & glass under influence of medium used.
International Pharmacopoeias.  The amount of acid to neutralize released
 In USP, used only for Soda-lime glass. alkali from surface is estimated.
 Containers are filled with test solution &  Methyl red is used to determine end point.
exposed to the test conditions.
 Glass wares may pass this test more easily
because surface layer is smooth & Creative.

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STATISTICAL
INTERPRETATION
OF QUALITY
CONTROL CHARTS
DURING
MANUFACTURING
PROCESSES

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Statistic
The collection, organization and interpretation of
data.
Quality
The International Organization for Standardization
(ISO) defines quality as ―the degree to which a set
of inherent characteristics fulfills requirements‖
Other definition is:
―The degree to which a product meets the
requirements of customer‖
Quality control
Quality control (QC) is a procedure or set of
procedures intended to ensure that a manufactured
product or performed service adheres to a defined
set of quality criteria or meets the requirements of
the client or customer.
Quality control is the regulatory process through
which we measure actual quality performance.
Levels of quality control
There are three levels of quality control
(1) Supplier level quality control
It deals with the quality of input materials i.e.
quality of raw materials
(2) In-process quality control level
It deals with the quality of partially completed
products i.e. quality during manufacturing process
(3) Customer level quality control
It deals with the quality of finished products
Quality Assurance
Quality Assurance is a system of activities whose
purpose is to provide an assurance that the overall
quality control is in fact being done effectively.
Quality Assurance: All those planned or systematic
actions necessary to provide confidence that a
product or service will satisfy given needs.
Statistical quality control
(SQC)
These are set of statistical tools used by quality
professionals to evaluate organizational quality.
Statistical quality control can be divided into three
broad categories:
1. Descriptive statistics
2. Statistical process control (SPC)
3. Acceptance sampling
Descriptive statistics
Statistics used to describe quality characteristics and
relationships, Included are statistics such as the
mean, standard deviation, the range, and a measure
of the distribution of data.
(a) Mean (average)
The arithmetic average, or the mean, is a statistic
that measures the central tendency of a set of data.
∑
To compute the mean we simply sum all the
observations and divide by the total number of
observations. The equation for computing the mean
is
Where
x = the mean
xi = observation i, i 1, . . . , n
n = number of observations
(b) Range
It is the difference between the largest and smallest
observations in a set of data.
(c) Standard deviation
It is a Statistic that measures the amount of data
dispersion around the mean.
It is a measure of average spread around the mean.
√∑ ( )

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The equation for computing the standard deviation

is

Where
 = standard deviation of a sample
x = the mean
xi = observation i, i 1, . . . , n
n = the number of observations in the sample

Small values of the range and standard deviation

mean that the observations are closely clustered
around the mean.

Large values of the range and standard deviation Positive skewness

mean that the observations are spread out around
the mean. If the data is skewed toward right than it‘s called
positive skewness. A long right tail in graph.
Mean > Median / Mode = Positive Skeweness

Negative skewness
If the data is skewed toward left than it‘s called
negative skewness. A long left tail in graph.
Mean < Median / Mode = Negative Skeweness

(d) Distribution of Data

A descriptive statistic used to measure quality
characteristics is the shape of the distribution of
the observed data.

Symmetric distribution
When a distribution is symmetric, there is the same
number of observations blew or above the mean.
Kurtosis:
This is what we commonly find when only normal
variation is present in the data. Kurtosis provides the visual estimation of variance
in a sample. It is a measure whether the data is peak
Skewness distribution or flat related to the normal distribution.
When a disproportionate number of observations
Leptokurtic
are either above or below the mean, the data has
skewed distribution. Kurtosis value greater than two is leptokurtic. It‘s
sharper than the normal distribution; values are
concentrated around the mean and little variance.

Platykurtic
Kurtosis for the negative number more than -1 is
called platykurtic distribution. It‘s flatter than the
normal distribution; values are spread out wider
from the mean and greater variance.
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Differences between symmetric and
skewed distributions
2) Statistical process control (SPC)
Statistical tool that involves inspecting a random
sample of the output from a process and deciding
whether the process is producing products with
characteristics that fall within a predetermined
range. SPC answers the question of whether the
process is functioning properly or not.
We use different types of control charts in statistical
process control.
3) Acceptance sampling
The process of randomly inspecting a sample of
goods and deciding whether to accept the entire lot
based on the results. Acceptance sampling
determines whether a batch of goods should be
accepted or rejected.
Adnan Sarwar Chaudhary
Control Charts
A control chart (also called process chart, Shewart
chart or quality control chart) is a graph that shows
whether a sample of data falls within the common
or normal range of variation.
The most commonly used tool for monitoring the
production process is a control chart.
Components of control chart
There are five components of a control chart:
X axis
The x axis represents samples, sequence or time.
Y axis
The y axis represents the quality characteristic that
is being monitored i.e. weight of tablet, volume of
injectable filled or values or results to be monitored.
Center line (CL)
The center line (CL) of the control chart is the
mean, or average, of the quality characteristic that
is being measured.
Control limits
Upper control limit (UCL) is the maximum
acceptable variation from the mean for a process
that is in a state of control.
LCL=mean-3 * sigma /n (1/2)
Lower control limit (LCL) is the minimum
acceptable variation from the mean for a process
that is in a state of control.
UCL=mean+3 * sigma /n (1/2)
Data points
The upper and lower control limits on a control
chart are usually set at ±3 standard deviations from
the mean. If we assume that the data exhibit a
normal distribution, these control limits will capture
99.74 percent of the normal variation.
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Control limits can be set at 2 standard deviations vi. Useful to highlight inter or intra batch
from the mean. In that case, control limits would variation
capture 95.44 percent of the values.

Control limit can be set at ±1 standard deviation

Types of Control Charts
from the mean. In the case, control limits would Control charts can be divided into two groups:
capture 68% of the values.
1. Charts for variables
2. Charts for attributes

(1) Control chart for variables

Control charts for variables monitor characteristics
that can be measured and have a continuous
scale, such as height, weight, volume, or width.

Control charts for variable are of following types:

1. x chart
2. R chart
3. S chart
4. S2 chart

a) Mean (x-Bar) Charts

A mean control chart is often referred to as an x-bar
chart. It is used to monitor changes so that a mean
value of a process can be obtained.

Center line and control limits are calculated as

CL = x =
Importance of quality control chart
UCL = x z σx
1. Proven technique for improving
productivity. LCL = x - z σx
2. Effective in defect prevention.
3. Prevent unnecessary process adjustment. Where
4. Provide diagnostic information. K = no of sample mean
5. Provide information about process Z = standard normal variable (2 for 95.44%
capability. confidence, 3 for 99.74% confidence)
6. As aid in controlling and analyzing physical, σx = standard deviation of the distribution of sample
chemical, analytical and biological means, computed as
parameters of production: σx =
√
i. Weight variation of tablets
ii. Thickness of tablets σ = population (process) standard deviation
iii. Volume of filled liquid in a container n = no of observations
iv. The number of defects or %age defects in
parenteral products Another way to construct the control limits when σ
v. The number or fraction of defects in the is not known is to use the sample range as an
sample of packages estimate of the variability of the process.

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In this case control limits would be constructed as
follows:
UCL = x + A2 R
LCL = x - A2 R
Where
X = average of sample mean
R = average range of samples
A2 = factor for x chart
b) Range (R) Charts
Range (R) charts are another type of control chart
for variables. Range charts monitor the
dispersion or variability of the process. Where-as
x-bar charts measure shift in the central tendency of
the process.
The method for developing and using R-charts is
the same as that for x-bar charts. The center line of
the control chart is the average range, and the upper
and lower control limits are computed as follows.
CL = R
UCL = D4 R
LCL = D3 R
Where
R = average range
D4 and D3 = factors for R chart
S Chart
In this chart, the sample standard deviations are
plotted in order to control the variability of a
variable.
S 2 Chart
In this chart, the sample variances are plotted in
order to control the variation of a variable.
(2) Control charts for attributes
A control chart for attributes is used to monitor
characteristics that have discrete values and can
be counted rather than measured. Often they can
be evaluated with a simple yes or no decision.
Examples include color, taste, or smell of product.
Control charts for attributes are of following types:
1. P chart
Adnan Sarwar Chaudhary
2. C chart
3. U chart
4. Np chart
1) P-Charts
Control chart that monitors the proportion of
defects in a sample is called P-chart.
P-charts are appropriate when both the number of
defectives measured and the size of the total sample
can be counted.
The computation of the center line as well as the
upper and lower control limits is similar to the
computation for the other kinds of control charts.
The center line is computed as the average
proportion defective in the population.
To construct the upper and lower control limits for a
p-chart, we use the following formulas,
CL = P
UCL = P zσp
LCL = P - zσp
Where
z = standard normal variable
P = the sample proportion defective
σp = standard deviation of the average
proportion defective calculated as
´( ´)
σp = √
2) C-CHARTS
A control chart used to monitor the number of
defects per unit.
Unit may be time, space or distance Examples are
the number of returned meals in a restaurant, the
number of trucks that exceed their weight limit in a
month.
C-charts count the actual number of defects.
However, we cannot compute the proportion of
complaints. Control limits are calculated as
CL = c
UCL = c z√
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LCL = c - z √ When? We say process is out of

Where control
c = average number of defects Process will be out of control when:
z = standard normal variable  A plot of data reveals that one or more
U chart samples fall outside the control limits.
 In this chart we plot the rate of defectives,  9 points in Zone C or beyond (on one side of
that is, the number of defectives divided by central line).
the number of units inspected (the n; e.g.,  2 out of 3 points in a row in Zone A or
feet of pipe, number of batches). beyond.
 4 out of 5 points in a row in Zone B or
 Unlike the C chart, this chart does not
beyond.
require a constant number of units, and it
 15 points in a row in Zone C (above and
can be used, for example, when the batches
below the center line).
(samples) are of different sizes.
 8 points in a row in Zone B, A, or beyond,
Np chart on either side of the center line (without
 In this chart, we plot the number of points in Zone C).
defectives (per batch, per day, per machine)  6 points in a row steadily increasing or
as in the C chart. decreasing.
 However, the control limits in this chart are  14 points in a row alternating up and down.
not based on the distribution of rare events,
but rather on the binomial distribution.
 Therefore, this chart should be used if the
occurrence of defectives is not rare (e.g.,
they occur in more than 5% of the units
inspected).
 For example, we may use this chart to
control the number of units produced with
minor flaws

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Comparison b/w control charts by

variables and attributes
Variables control chart Attributes control
charts

Monitor quality Monitor quality

characteristics that can characteristic that can be
be measured counted rather than
measured

Based on continuous Based on discrete

values values

Uneconomical Economical

Cha Process Process Process Size Less easily More easily

rt observation observatio observati of understandable understandable
ns ons shift
relationsh type to
ips dete
more sensitive or Less sensitive or
ct efficient efficient
and Quality Independe Variables Larg
R characteristic nt e (≥
char measurement 1.5σ Process capability
t within one )
subgroup The ability of a production process to meet or
and Quality Independe Variables Larg
s characteristic nt e (≥ exceed preset specifications. This is called process
char measurement 1.5σ capability.
t within one )
subgroup Product specifications often called tolerances are
p- Fraction Independe Attributes Larg Preset ranges of acceptable quality characteristics.
char nonconformin nt † e (≥
Process capability involves evaluating process
t g 1.5σ
within one ) variability relative to preset product specifications
subgroup in order to determine whether the process is capable
np- Number Independe Attributes Larg of producing an acceptable product.
char nonconformin nt † e (≥
t g 1.5σ Measuring Process Capability (Capability of
within one )
process =Cp)
subgroup
c- Number of Independe Attributes Larg
nonconforman nt † e (≥
Simply setting up control charts to monitor whether
char
t ces within 1.5σ a process is in control does not guarantee process
one subgroup ) capability. To produce an acceptable product, the
u- Nonconforma Independe Attributes Larg process must be capable and in control before
char nces per nt † e (≥
t unit within 1.5σ production begins.
one subgroup )
Process capability is measured by the process
capability index, Cp, which is computed as the

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ratio of the specification width to the width of the

process variability:
Where
Specification width is the difference between the
Upper Specification Limit (USL) and the Lower
Specification Limit (LSL) of the process.
Interpretation of Cp value
There are three possible ranges of values for Cp
that also helps us interpret its value:
Cp = 1: A value of Cp equal to 1 means that the
process variability just meets specifications, as in
Figure (a). We would then say that the process is
minimally capable.

Process width is computed as 6 standard deviations Cp < 1: A value of Cp below 1 means that the
(6σ) of the process being monitored. process variability is outside the range of
specification, as in Figure (b). This means that the
Another measure for process capability is used process is not capable of producing within
more frequently: specification and the process must be improved.
( ) Cp > 1: A value of Cp above 1 means that the
process variability is tighter than specifications and
the process exceeds minimal capability, as in Figure
Where

µ = the mean of the process

σ = the standard deviation of the process

To use this measure, the process capability of each

half of the normal distribution is computed and the
minimum of the two is used.

Factors for three-sigma control limits of and -

charts
Factor for Computing Central Line and Three Sigma Limits
Observation A A2 D1 D2 D3 D4 A3 B3 B4 d2 c4
in Sample
(n)
2 2.121 1.880 0 3.686 0 3.267 2.659 0 3.267 1.128 0.7979
3 1.732 1.023 0 4.358 0 2.574 1.954 0 2.568 1.693 0.8862
4 1.500 0.729 0 4.698 0 2.282 1.628 0 2.266 2.059 0.9213
5 1.342 0.577 0 4.918 0 2.114 1.427 0 2.089 2.326 0.9400
6 1.225 0.483 0 5.078 0 2.004 1.287 0.030 1.970 2.534 0.9515
7 1.134 0.419 0.204 5.204 0.076 1.924 1.182 0.118 1.882 2.704 0.9594
8 1.061 0.373 0.388 5.306 0.136 1.864 1.099 0.185 1.815 2.847 0.9650
9 1.000 0.337 0.547 5.393 0.184 1.816 1.032 0.239 1.761 2.970 0.9693
10 0.949 0.308 0.687 5.469 0.223 1.777 0.975 0.284 1.716 3.078 0.9727
11 0.905 0.285 0.811 5.535 0.256 1.744 0.927 0.321 1.679 3.173 0.9754
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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

12 0.866 0.266 0.922 5.594 0.283 1.717 0.886 0.354 1.646 3.258 0.9776
13 0.832 0.249 1.025 5.647 0.307 1.693 0.850 0.382 1.618 3.336 0.9794
14 0.802 0.235 1.118 5.696 0.328 1.672 0.817 0.406 1.594 3.407 0.9810
15 0.775 0.223 1.203 5.741 0.347 1.653 0.789 0.428 1.572 3.472 0.9823
16 0.750 0.212 1.282 5.782 0.363 1.637 0.763 0.448 1.552 3.532 0.9835
17 0.728 0.203 1.356 5.820 0.378 1.622 0.739 0.466 1.534 3.588 0.9845
18 0.071 0.194 1.424 5.856 0.391 1.608 0.718 0.482 1.518 3.640 0.9854
19 0.688 0.187 1.487 5.891 0.403 1.597 0.698 0.497 1.503 3.689 0.9862
20 0.671 0.180 1.549 5.921 0.415 1.585 0.680 0.510 1.490 3.735 0.9869
21 0.655 0.173 1.605 5.951 0.425 1.575 0.663 0.523 1.477 3.778 0.9876
22 0.640 0.167 1.659 5.979 0.434 1.566 0.647 0.534 1.466 3.819 0.9882
23 0.626 0.162 1.710 6.006 0.443 1.557 0.633 0.545 1.455 3.858 0.9887
24 0.612 0.157 1.759 6.031 0.451 1.548 0.619 0.555 1.445 3.895 0.9892
25 0.600 0.153 1.806 6.056 0.459 1.541 0.606 0.565 1.435 3.931 0.9896

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Adnan’s Pharmaceutical Quality Management Adnan Sarwar Chaudhary

Adnan Sarwar Chaudhary

deenasaan@gmail.com

03041038728

Other Books
1. Basic Pharmacognosy by Hafiz Abdul Khaliq
2. Advance Pharmacognosy by Hafiz Abdul Khaliq
3. Adnan's Biostatistics by Adnan Sarwar Chaudhary
4. Adnan's Computer For Pharmacist by Adnan Sarwar Chaudhary & Saad
Muhammad Rustam
5. Adnan's Pharmaceutical Instrumentation by Adnan Sarwar Chaudhary &
Saad Muhammad Rustam
6. Adnan‘s Professional English by Adnan Sarwar Chaudhary & Saad
Muhammad Rustam
7. Nida‘s Pathology by Adnan Sarwar Chaudhary & Nida Rehman Alvi NB.

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