Quality Control

of Crude Drugs

A variety & sheer No. of plants with therapeutic activity is

astonishing.
A pharmacist must capable of knowing all the reliable
information on the quality, efficacy, safety and rational
uses of all the herbal formulations.
Herbal Preparation:
Fresh Juice, paste/power, Decoction, Cold/Hot infusion

Quality STDS of Herbal Products

Represents a number unique problem
Because of nature of the ingredients present
These are complex mixture of 2o metabolites
Considerable depending on the environmental and
genetic factors
Which are responsible for therapeutic effects

 Hence standards are used

To determine genuine drug
Methods used for adulteration
Detection of adulteration

Types
Structural STDs
Analytical STDs
STDs relating physical constants

Structural STDs
Certain parts of the plants mixes
Carelessness during the process of collection or preparation or
both
The materials with mixed accidently or deliberately called foreign
organic matter
Standard commonly
2% as per monograph

Analytical STDs
Concerned with the constituents of the
materials present within the herbal drugs
Std 1) Definite chemical Entity
Presence of minimum amount of useful
constituents
e.g. Strychnine, Quinine
2) Mineral Matter content
Estimation of ash values
Total ash
Acid soluble Ash

 STDs relating physical constants

Properties like Density, Viscosity, Ref. Index, Optical
rotation

Factors relating to quality of herbal drugs

1)Authentication & reproducibility of herbal ingredients
Accurately identified by macroscopic & Microscopical
characters are compared with authenticated material
Referred
Binominal Latin names of genus & species
Permitted synonyms should be used

2) Intra/Inter species variation in Plants:

1o & 2o metabolites also varies
Controlled by genetically

3) Environmental Factors
Climate, rainfall, altitude etc.

4) Plant parts used

Constituents vary between plant parts

5) Time of harvesting
Optimum time Constituents concentrates

6) Post harvesting factors

Storage, transport
Improper- Contamination

7) Contaminants of Herbal Ingredients

Free from insects, other animal matter and excreta
determined by ash values

8) Pesticides, Fumigants & other toxic metals

DDT, Organophosphate, Carbamates in cultivated crops
Pesticide Ethylene oxide, methyl bromide
- Pb, Cd, Hg etc.
- Contaminants of herbal ingredients

Salient features of who

guidelines (1991)
Quality control of crude drug material, plant
Preparation & finished products
Stability assessment and shelf life
Safety Assessment Toxicological studies
Efficacy Assessment Biological studies
Parameters in Quality control
Sampling Procedures
Morphological Examinations
Microscopical
Evaluation
Standardization
Parameters
Phytoconstituents and analysis

Sampling Procedures
Assessment of the quality of herbal drugs is
directly dependent on the selection of sample
for exam:
No. of packages in
shipment
1-10

No. of packages to be
sampled
1-3

10-25

2-4

25-50

3-6

50-75

6-8

75-100

8-10

Determination of Foreign
Matter

Prescribed the permissible limit for the other

parts of the pits/the organic matters.
Animal excreta, moulds, insects or other animal
contamination
Procedure
Specific quantity spread on a thin layer of
paper
Examine by using magnifying lens
The percentage is recorded
WHO guidelines:
Leaves, flowers, seeds & Fruits 250mg
Roots, Rhizomes & Barks 500gm
Cut Medicinal Plant Materials 50gm

Morphological Examinations
Organoleptic Evaluation

Colour
Size
Odour
Taste

Surface characteristic
Texture
Fracture

Macromorphography/ Gross Morphology

Whole drug- studies

Aerial stem
Arrangement of leaves
Description of flowers
Description of Fruits
Unorganized drugs

Cytomorphological evaluation
Basic cell types: Parenchyma, Sclerenchyma, Collenchyma,
Xylem, Phloem, Epidermis
Cell Inclusions CaCO3, Starch, Oil cells

Microscopical Evaluation
Instruments for Microscopical Study:
Camera Lucida, Photomicrography
Modified light microscopy, Polarizing microscopy
Phase contrast microscopy, Electron microscopy
UV microscopy

PPN of crude drug sample

Clearing reagents
Chloral Hydrate soln.
HCl
Caustic soda

 Micro chemical Testing

Phloroglucinol stains lignified cells

Iodine Starch
Picric Acid Aleurone grains
Sudan III solution Secretary cells
Ruthenium red Mucilage

Micro chemical Precipitate

Atropine + Wagners reagent small dark rods
Ephedrine + Krauts reagent Tree shaped crystals

Micro chemical Sublimation

Cineol + Resorcinol in 15-30% acetic with
Crystalline additional product.

Quantatitive Microscopy
Find out & identify drugs either in fresh or in dry
condition.
Parameters:

Vein islet No. & vein termination nos.

Stomatal index & Stomatal Number
Palisade Ratio
Microscopic measurements
Determination of length/width/diameter of tissues,
vessels, fibres, vittae tubes, oil glands, starch grains.
Determination of no. of size of uniform tissues in one
mg. of the given drug powder.
Constant for particular species of pH independent of
its age & size.

Vein Islet Number

Average no. of V.I. present in 1 sq.mm of the
central part of the lamina, between the midrib &
margin.
Procedure

Cut portion
Cleaning solution
Mount in glycerin
Arrange camera lucida & drawing board
Stage m Calibration
Stage mm is replaced by the cleared leaf section on
the slide
Count the islets
4 trials to be done

Vein Termination Number

Average no. of vein terminals present in 1
sq.mm of the same lamina, between the midrib
& margin
Procedure:

Cut portion
Cleaning solution
Mount in glycerin
Arrange camera lucida & drawing board
Stage m Calibration
Stage mm is replaced by the cleared leaf section on
the slide
Count the islets
4 trials to be done

Stomatal Number & Stomatal

Index

Stomatal No. No. stomata/Sq.mm of

epidermis
Stomatal Index the % proportion of
stomata and epidermal cells with stomata
Constant
S.I =
S
x 100
E+S
S No. of stomata per unit area
E No. of epidermis in same unit area

Palisade Ratio
Average no. of palisade cells present
under on epidermal cells
Determined in fine powder
Diagnostic value constant
Procedure
Pieces from base, middle & apex of young as
well as adult leaves
Clearing solution
Leaves become transparent
Camera Lucida

Lycopodium Spore Method

T.E. Wallis
Determination of % purity of drug & no. of grains present
in the drug sample
Consist of the common club moss
Lycopodium clavatum

Family: Lycopodiaceal
Orders: Pteridophytes
Each spore tetrahedral shape is uniform in size
94,000 spores/mg constant

Procedure:

Drug powder: Lycopodium powder (2:1)

Mix with olive oil
Make uniform suspension
Place drop of suspension on slide
Put cover slip
Examine under microscope

% Purity of sample drug = N x W x 94,000 x 100

SxPxM
N No. of ginger starch grains in 25 fields
W Wt. of Lycopodium Powder in mg.
S No. of spores
P No. of characteristics particles/mg of sample
M Wt. of sample in mg.
Determination of lg/wd/diameter of tissues

Calibrate the eye piece m meter using stage mm and calculate the factor
Powder + (Phloroglucinol + Conc. HCl)
Mount in dil. Glycerine
Observe under low power
Measure the width
Calculate the values of fibres & multiply them by factor

Swelling factor
Mucilage in the epidermis
1gm Isaphagol seed + 20ml H2O 24 hrs
Measure

Standardisation Parameters
Determination of Solvent Extractive values:
Determines the amount of active constituents in a given amount
of medicinal plant material when extracted with solvents
Employed for material for which no. chemical (or) biological
assay method exist

Water soluble extractive:

Method I:
5 gm air dried drug + 100ml H2O
Macerated for 23 hours
Shaking frequently during 1st 6 hours & stand for 18 hrs
Filter
Evaporate 25ml of the filtrate to dryness
Dry at 105oC weigh
% of water soluble extractive with reference to air dried drug
have to be calculated

Method I:
5 gm drug + 50ml of H2O at 80oC in stoppered flask
Shake well, stand for 10 mts
Add 2gm Kieselguhr
Filter
5ml of the filtrate
Evaporate, dry for 30 mts
Dry in oven for 2 hrs
Weight the residue
Calculate the % of water soluble extractive

2) Determination of alcohol soluble extractive

5gm of drug +100ml of ethanol (specified strength)
24 hrs
Shaking frequently during the 1st 6 hrs & Std for 18 hrs
Filter
Evaporate 25ml of the filtrate dry at 105oC & weigh
% of ethanol soluble extractive has to be calculated

3) Solvent Hexane soluble extractive:

2 gm drug Extract with solvent hexane for 20 hrs
Evaporate spontaneously
Dried over P2O5 for 18 hrs
Weight & Heat the extract
Dry at 105oC
Weight, loss in wt represents the volatile portion of extract

5) Non volatile ether soluble extractive:

Similar to volatile ether soluble extractive,
weight of extract after drying at 105 oC
represents the non volatile portion of the
extract.
II) Determination of Ash Values:
Represents their non-volatile inorganic
components (metallic salts & Silica)

1) Total Ash:
Total amount of material produced after
complete incineration of the ground drug at as
low temp (450oC) to remove all the carbons
Physiological ash + Non-physiological ash
(Plant)
(Sand & Soil)
Total Ash
2-3 gm of drug in patient (or) silica dish
Incinerate at a temperature not exceeding 450 oC
Cool & Weight

2) Acid insoluble Ash (I.P. 1996)

Residue obtained after extracting the total ash
with HCl.
Calculate with reference to 100gm of drug
Method:
Ash + 25ml 2m HCl
Boil for 5 mts
Collect the insoluble matter
Wash with hot H2O
Ignite, cool & weight

3) Water soluble ash:

Difference in weight between the total ash & the residue
obtained after treatment of total ash with H 2O.
Part of the total ash content which is soluble in H2O.
Method: I.P. 1996
Boil the ash with 25ml H2O
For 5mts
Collect the insoluble matter
Wash with H2O
Ignite for 15mts at temp. not exceeding 450oC
Subtract the weight of the insoluble matter from the weight
of the ash
Difference represents H2O soluble ash
Calculate with reference to air dried drug

4) Determination of Total Solids:

Residue obtained when the treated amount of the preparation
is dried to constant weight under specified condition.
Method:
Weight the drug
Evaporate, heat on H2O bath
Dry to constant weight at 105oC
Cool weight
5) Determination of crude fibre:
Denotes the measurements of the content of cellulose, lignin
& cork cell in the plant tissue (more resistant part of plant cell)
Indicates adulteration with woody tissues likes kernels
Other than ash which can not be dissolved in H 2O & can not
be digested by boiling with H2SO4 (or) with NaOH.

Method:

2 gm drug + ether
Extraction
Add 200ml 1.25% H2SO4
Boil for 30 mts under reflux
Filter
Wash the residue with boiling H2O until free from acid.
Take the residue in flask with 200ml of boiling 1.25% NaOH Solution*
Boil again for 30 mts
Filter
Residue is washed with boiling H2O until neutral, dried at 110oC to
constant wt.
Incinerate to constant wt
Difference between the weight of the dried residue & that of the
incinerated residue represents the weight of the crude fibre

VI) Determination of Moisture Content:

Excess moisture cause breakdown of important
constituents by enzymatic activity & encourage
the growth of yeast & fungi during storage.
Determined by loss on drying, volumetric
azeotropic distillation method, Karl fischer
method.
A) Loss on drying:
Major loss in wt. due to H2O & V.O.L it present
Method: Prescribed qty of the substance is dry to
constant mass for specified time in dessicator
over P2O5 (or) in oven.

B) Azeotropic distillation method: (Tolura distillation)

Organic solvent (toluene, benzene, xylene) forms a
azeotropic mixture with H2O Present in the crude drug.
When the drug is heated, org-solvent & H2O is distilled
together which is collected in graduated tube of the
apparatus. Water forms a bottom layer can be directly
lead after complete distillation (Deavestark apparatus)
C) Karl fischer Method:
Applicable to very small qty of moisture
Reagent Iodine + SO2 + Pyridine in dry CH3OH
I2 is reduced by SO2 is presence of H2O causing the loss
of dark brown colour of the reagent.

VII. Determination of Essential oils in crude drugs:

By hydro distillation (or) steam distillation
Clavengers apparatus is used for estimation
During hydro distillation, addition of 5-10% glycerin
cause better yield of V. oil (4-5 hrs)
Collected Volatile oil measured from graduated receiver
VIII. Determination of Bitterness value:
Bitters have strong bitter taste, employed therapeutically
as appetizing agents.
Bitterness stimulates the gastrointestinal secretions
Bitterness value determined by comparing the threshold
bitter conc. Of an extract of the materials with that of
dilute soln. of quinine HCl (1 gm is 2000ml)

IX. Determination of Haemolytic activity:

Saponins characterised by their frothing property
Cause haemolysis when added to suspension of blood.
(produce change in erythrocyte membranes, causing
haemoglobin to diffuse into the surrounding medium)

X. Determination of swelling Index:

E.g: Seals (Plantago ovata)
Measuring the volume of mucilage produced in 24 hrs
from 1 gm of the seeds. This is known as swelling factor.
Swelling Index: Volume in ml taken up by the swelling of
1gm of plant material under specific condition.

XI. Determination of Foaming index:

Saponins
The foaming ability of an aqueous decoction of
plant materials & their extracts is measured in
terms of foaming index.
XII. Determination of Tannins:
Gold beaters test (or) tanning test:
Capable of turning animals hides into leather by
binding proteins to form water soluble
substances that are resistant to proteolytic
enzymes.

XIII. Residual Analysis:

A) Pesticide Residues:

Intended to be used for protection of plants

against certain damaging influences such as
insects, fungal diseases, weeds, rodents
Many developed countries have issued
restricting regulations concerning the use of
pesticides.
B) Heavy metals analysis:

Such as arsenic, head, cadmium, mercury

should be analysed which are toxic to human.
Achieved by means of chemical analysis, atomic
absorption method, colorimetric method.

 Limits for pesticide residues, arsenic and heavy

metals can be
Maximum residue limits
= ADI x BW x Extraction factor
MDI x safety factor x 100
ADI Acceptable daily intake as per WHO
BW Body weight
MDI Mean daily intake of drug
XIV.

Determination of Radioactivity:

Medicinal plants may be exposed to radio

nucleides, therefore they should be tested for
residual levels of radioactivity.

XV. Determination of Microbial

contamination
The microbial quality of medicinal products
has attracted much attention because
which leads to clinical infection.
The limits are: 10 x Microorganism per g
Aerobic bacterial count should be within 102 108 cfu/g
Coliform bacterial count should be within 102 108 cfu/g
Fungal count should be within 102 108 cfu/g

REFERENCES
Trease and Evans Pharmacognosy 14th
Edition, WB Saunders Co., Ltd., London, 1996.
Indian Pharmacopoeia, 4th Edn, Ministry of
Health and Welfare, New Delhi, 1996.
Wallis, T.E.1985, Text book of Pharmacognosy,
Fifth Edition, CBS publishers, New Delhi, India.
WHO/Pharm/92-559/Quality central methods
for medicinal plant materials, 1992.
Edward 8, Claus, Varro E, Tyler, JR,
Fifth Edition, 1965.