THESE R TEST RELATED TO QUALITY CONTROL METHOD FOR EVALUATION OF CRUDE DRUG AS PER WHO

1. COMPENDIAL METHODS FOR
EVALUATION OF CRUDE DRUG AND
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HERBAL FORMULATION
Prepared by: Jagani Nayan M.
M. Pharm (sem-II) Q.A.
Guided by : Mr. Jignesh S. Shah,
Asst. Professor,
Q. A. department.
1
Department of Quality Assurance
SJTPC - RAJKOT

2.  INTRODUCTION
 Definition: A crude drug is naturally occurring, unrefined substance
derived from organic or inorganic sources such as plant,
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animal, bacteria, organs or whole organisms intended for
use in the diagnosis, cure, mitigation, treatment, or
prevention of disease in man or other animals.
 The term “herbal drugs” denotes plants or plant parts that have been
converted into phytopharmaceuticals by means of simple processes
involving harvesting, drying, and storage.
 Herbalism is a traditional medicinal or folk medicine practice based
on use of plant and plant extracts known as herbal medicine,
phytotherapy, herbology etc.
 Following are the Q. C. Parameter for evaluation of crude drug and
herbal formulation :
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3.  PARAMETERS FOR EVALUATION OF
HERBAL DRUGS
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(A) PHYSICAL PARAMETERS
 Viscosity,  Solubility,
 Volatile matter,  Moisture content,
 Density,  Specific gravity,
 Refractive index,  Optical rotation,
 Hemolytic activity,  Bitterness value,
 Foaming index,  Swelling index,
 Melting point,  Ash value.
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4. (B) CHEMICAL PARAMETER
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 Detection of alkaloids.
 Detection of carbohydrates and glycosides.
 Detection of phytosterols.
 Detection of fixed oils and fats.
 Detection of saponins.
 Detection of phenolic compounds and tannins.
 Detection of protein and free amino acids.
 Detection of gums and mucilage.
 Detection of volatile oils.
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5. (C) BOTANICAL PARAMETER
 Microscopical parameters.
 Macroscopical parameters.
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(D) BIOLOGICAL / TOXICOLOGICAL PARAMETER
 Determination of pesticides.
 Determination of arsenic and heavy metals.
 Determination radioactive contamination.
 Determination of aflatoxins.
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6.  General considerations:
 The metric system is used throughout. All temperatures are
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expressed in degrees Celsius (°C). Tests are normally carried out
at room temperature (between 15 and 25°C, or up to 30°C in
some climatic zones).
 Any glassware used in the tests should be of suitable quality.
Graduated and volumetric vessels should be calibrated at room
temperature.
 When a water-bath is referred, a bath containing boiling water
(about 100°C) is to be used, unless a specific water temperature
is given.
 Unless otherwise specified, all solutions indicated in the tests are
prepared with distilled or demineralized water of adequate
purity.
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7.  Determination of Foreign Matter:
Any organism, part or product of an organism, other than that named in
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
the specification and description of the plant material concerned.
 Material not adhering to the medicinal plant materials, such as soil,
stones, sand, and dust.
 Sample size:
o Plant material Sample size
o roots, rhizomes and bark 500 g
o leaves, flowers, seeds 250 g
o cut medicinal plant materials
o (average weight of each fragment less than 0.5 g) 50 g
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8.  Recommended procedure:
 Weigh 100 –500 g of the drug sample to be examined, or the
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minimum quantity prescribed in the monograph, and spread it
out in a thin layer. The foreign matter should be detected by
inspection with the unaided eye or by the use of a lens (6x).
 Separate and weigh it and calculate the percentage present.
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9.  Determination of Pesticides:
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 Pesticides are chemicals derived from synthetic and natural sources
which are effective in small concentrations against pest.
 Even though there are no serious reports of toxicity due to the presence
of pesticides and fumigants, it is important that herbs and herbal
products are free of these chemicals or at least are controlled for the
absence of unsafe levels.
 Herbal drugs are liable to contain pesticide residues, which accumulate
from agricultural practices, such as spraying, treatment of soils during
cultivation, and administering of fumigants during storage. However, it
may be desirable to test herbal drugs for broad groups in general, rather
than for individual pesticides.
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10.  Classification of pesticides (according to pest they control):
A) Fungicides: prevents plant from diseases caused by phyto pathogenic
fungi and used as soil or seed fungicidal disinfectants.
Types:
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a. disinfectants for seed: parathion, carboxins.
b. disinfectants for soil:
c. Leaf fungicides: colloidal sulphur, barium sulphide,
dithiocarbamates.
B) Herbicides: ‘weed killers’ which are used for destroying the unwanted
plants.
Types:
a. Total herbicides
b. Selective herbicides (widely used)
c. Water weed killers
d. Harvesting aids
Eg.: carbamates, urea derivatives, triazines , quarternary ammonium
compounds.
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11. C) Insecticides: used to protect the crop from insects.
Types:
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a. Inorganic insecticides:
arsenic compounds (lead arsenate)
flourine compounds (sodium floride).
b. Orgenic insecticides: nicotine, pyrethrum
D) Others like nematocides, rodenticides, bactericides.
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12.  Method:
20-50 gm powder + acetonitrile –water mixture
blend for 5min at high speed
& filter
take filtrate & transferred in 1000ml searating funnel & add 100ml of
light petroleum ether.
shake, 1-2min.
Add 10ml NaCL(40%) +600ml water
shake vigorously for 30-45sec.
Petroleum ether layer is collected & washed with water thrice.
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13. Cont…
This is treated with anh. Sodium sulphate.
extract is subjected to column chromatography.
i. column packing: activated fluorosil
ii. eluted with: petroleum ether
iii. 3 fractions of 200ml are collected.
a.1st elute contains chlorinated pesticides.
b. 2nd elute contains dieldrein.
c. 3rd elute contains malathion.
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14. cont…
iv. The elutes are concentrated to 10ml & used for TLC or HPTLC.
v. Particulars for TLC:
- standard samples: in petroleum ether.
- adsorbent: pre-coated silica gel GF254 plate (10*20cm) of
uniform thickness.
- solvent system: N-hexane – acetone.
- detection: under UV chamber.
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15.  Maximum limit of pesticide residues for herbal drugs:
 According to EP(1997) limits for pesticides calculated using following
formula:
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MRL= ADI * W____
MDI * (100 * safety factor)
 Where,
MRL: maximum residue limits(mg/kg)
ADI: acceptable daily intake =NOEL *(safety factor )
NOEL: non-observable effect level.
W: body weight(kg)
MDI: mean daily intake of drug.
 Safety factor: 1/100 to 1/2000.
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16.  When the herbal crude drug is used to prepare the extracts, tinctures
or other phytopharmaceutical formulations in which there is difference
in concentration of pesticide residues, so the MRL is calculated as :
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MRL= ADI * W * E____
MDI * (100 * safety factor)
where, E: the extraction co-efficient of the pesticide which depend on
method of preparation.
 The quantity of the pesticide transferred from raw material to the final
formulations depends on several factors like:
 Types and quantity of the solvents.
 Temperature.
 Duration of heating.
 Degree of contamination.
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17.  DETERMINATION OF HEAVY METAL: (AS PER WHO)
 Limit test for Arsenic
 Limit test for cadmium and lead
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18.  Metal:
Characterized by luster, malleability, high electric and thermal
conductivity; chemically form bases which can react with acids
 Heavy Metal: Metal of High Specific Gravity:
- Cadmium 112 - Lead 207
- Mercury 200 - Zinc 65 (Not so heavy)
- Aluminum 27 (Light) -Arsenic 75
- Selenium 79
 Heavy Metals – Sources:
 In general: Worldwide: associated with mining industry.
 Others:
- Environmental pollution,
- Accidental inclusion in processing,
- Contamination from containers. 18
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19.  Excess heavy metal accumulation in soil is toxic to humans and
other animals.
 Exposure to heavy metals is normally chronic.
(exposure over a longer period of time), due to food chain transfer.
 Acute (immediate) poisoning from heavy metals is rare through
ingestion or dermal contact, but is possible.
 Chronic problems associated with long-term heavy metal
exposures are:
Lead – mental lapse.
Cadmium – affects kidney, liver, and GI tract.
Arsenic – skin poisoning, affects kidneys and CNS.
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20.  LIMIT TEST FOR ARSENIC:
 Principle:
The amount of arsenic in the medicinal plant material is estimated by
matching the depth colour with that of a standard stain.
 Reactions:
H2AsO4 KI H3AsO3
Arsenic acid arsenious acid
H3AsO3 + 3H2 AsH3 + H2O
Arsenious acid arsine gas
AsH3 + mercuric bromide yellow stain
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21.  Preparation of the sample (by acid digestion):
Place35-70 g of coarsely ground material, accurately weighed, in a Kjeldahl
flask, capacity 800-1000 ml. +(10-25 ml) water + (25-50 ml) nitric acid +
carefully add (20 ml) sulphuric acid.
Heat
Gradually add nitric acid (~1000 g/l), drop by drop, until all the organic
matter is destroyed. a clear solution with copious vapours of sulfur trioxide
is obtained.
Cool
add (75 ml) water + (25 ml) ammonium oxalate.
Heat again until sulfur trioxide vapours develop.
Cool, transfer to a 250-ml volumetric flask and dilute to
volume with water. 21
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22.  Preparation of standard stain:
Add (10 ml) hydrochloric acid +
1 ml dilute arsenic As to 50 ml water.
The resulting solution, when treated as described in the general test
yields a stain on mercuric bromide paper.
standard stain (10 μg of As).
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23.  Method:
Take an aliquot (25-50 ml) of the test solution +1 g of potassium
iodide + l0 g of granulated zinc in the widemouthed bottle place
the prepared glass tube assembly quickly in position.
reaction 40 minutes,40’c temp.
Compare any yellow stain that is produced on the mercuric
bromide paper with a standard stain produced in a similar manner
with a known quantity of dilute arsenic.
 The contents of lead and cadmium may be determined by atomic
absorption spectrophotometry
Maximum amount in dried plant materials:
-lead: 10mg/kg. -cadmium: 0.3mg/kg. 23
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24.  Determination of Ash:
 The ash remaining following ignition of medicinal plant materials is
determined by three different methods which measure total ash, acid-
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insoluble ash and water-soluble ash.
 The total ash method is designed to measure the total amount of
material remaining after ignition. This includes both "physiological
ash", which is derived from the plant tissue itself, and "non-
physiological" ash, which is the residue of the extraneous matter (e.g.
sand and soil) adhering to the plant surface.
 Acid-insoluble ash is the residue obtained after boiling the total ash
with dilute hydrochloric acid, and igniting the remaining insoluble
matter. This measures the amount of silica present, especially as sand
and siliceous earth.
 Water-soluble ash is the difference in weight between the total ash and
the residue after treatment of the total ash with water.
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25.  Recommended procedures:
 Determination of Total Ash
 Incinerate about 2 to 3 g accurately weighed, of the ground drug in a silica
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dish at a temperature not exceeding 450º until free from carbon, cool and
weigh. If a carbon free ash cannot be obtained in this way, exhaust the
charred mass with hot water, collect the residue on an ashless filter paper,
incinerate the residue and filter paper, add the filtrate, evaporate to
dryness, and ignite at a temperature not exceeding 450º. Calculate the
percentage of ash with reference to the air-dried drug.
 Determination of Acid Insoluble Ash
 Boil the ash obtained above for 5 minutes with 25 ml of dilute hydrochloric
acid; collect the insoluble matter in a Gooch crucible, or on an ashless filter
paper, wash with hot water and ignite to constant weight. Calculate the
percentage of acid-insoluble ash with reference to the air dried drug.
 Determination of Water Soluble Ash
 Boil the ash for 5 minutes with 25 ml of water; collect insoluble matter in a
Gooch crucible, or on an ashless filter paper, wash with hot water, and ignite
for 15 minutes at a temprature not exceeding 450º. 25

26.  Determination of swelling index:
 Many medicinal plant materials are of specific therapeutic or
pharmaceutical utility because of their swelling properties, especially
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gums and those containing an appreciable amount of mucilage, pectin or
hemicellulose.
 The swelling index is the volume in ml taken up by the swelling of 1 g
of plant material under specified conditions. Its determination is based
on the addition of water or a swelling agent as specified in the test
procedure for each individual plant material (either whole, cut or
pulverized). Using a glass-stoppered measuring cylinder, the material is
shaken repeatedly for 1 hour and then allowed to stand for a required
period of time. The volume of the mixture (in ml) is then read.
 The mixture of whole plant material with the swelling agent is easy to
achieve, but cut or pulverized material requires vigorous shaking at
specified intervals to ensure even distribution of the material in the
swelling agent. 26

27.  MACROSCOPIC EXAMINATION
 Organoleptic evaluation of drugs refers to the evaluation of a drug by
colour, odour, size, shape, taste and special features including touch,
texture etc. Since the majority of information on the identity, purity and
quality of the material can be drawn from these observations, they are of
primary importance before any further testing can be carried out.
 For this purpose authentic specimen of the material under study and
samples of pharmacopoeial quality should be available to serve as a
reference.
 This evaluation procedure provides the simplest and quickest means to
establish the identity and purity and thereby ensure quality of a
particular sample.
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28.  If it is found to be devoid of or significantly different from the
specified sensory characters like colour, consistency, odour, etc., it
is considered as not fulfilling the requirements.
 However judgment based on the sensory characteristics like
odour, taste etc., may vary from person to person and time to time
based on individual's nature.
 So the description of this features are very difficult so that often
the characteristic like odour and taste can only described as
'characteristic' and reference made to the analyst's memory.
 No preliminary treatment is necessary for evaluating the sample
in this manner excepting the softening and stretching of the
wrinkled and contracted leaves and flowers etc.

29.  MICROSCOPIC EXAMINATION
 This allows more detailed examination of a drug and it can be used to
identify the organised drugs by their known histological character.
 This mostly used for qualitative evaluation of organised crude drugs in
entire and powdered forms and can be used to distinguish cellular
structure.
 For the effective results, various reagents can be used to distinguish
cellular structure.
eg. A drop of phloroglucinol and conc. Hcl gives red stain with lignin.
Mucilage is stained with rhuthenium red to give pink color.
 Following are the parameter evaluated by histological examination:
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30. a) Stomatal index : is the percentage which give the number of stomata to
total number of epidermal cells.
 Procedure : Place leaf fragments of about 5 × 5 mm in size in a test
tube containing about 5 ml of chloral hydrate solution and heat in a
boiling water-bath for about 15 minutes or until the fragments become
transparent. Transfer a fragment to a microscopic slide and prepare the
mount, the lower epidermis uppermost, in chloral hydrate solution and
put a small drop of glycerol-ethanol solution on one side of the cover-
glass to prevent the preparation from drying. Examine with a 40x
objective and a 6x eye piece, to which a microscopical drawing apparatus
is attached. Mark on the drawing paper a cross (x) for each epidermal
cell and a circle (o) for each stoma.
 It is calculated by following equation :
S. I. = S / E + S * 100
where, S = no. of stomata per unit area, 30
E = no. of epidernal cells in unit area.

31. b) Stomatal no.: is the average no. of stomata present per sq mm of
epidermal cell.
c) Palisade ratio : is the average no. of palisade cells beneath each
epidermal cell.
d) Vein islet & veinlet termination number : is the no. of vein islets &
veinlet termination per sq mm of the leaf surface midway between
midrib and margin.
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32.  Determination of Volatile Oil in Drugs
 Volatile oils are characterized by their odour, oil-like appearance and
ability to volatilize at room temperature. Chemically, they are usually
composed of mixtures of monoterpenes, sesquiterpenes and their
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oxygenated derivatives.
 Volatile oil is considered to be the "essence" of the plant material, and
are often biologically active, they are also known as "essential oils".
 The determination of volatile oil in a drug is made by distilling the
drug with a mixture of water and glycerin, collecting the distillate in a
graduated tube in which the aqueous portion of the distillate is
automatically separated and returned to the distilling flask, and
measuring the volume of the oil.
 The content of the volatile oil is expressed as a percentage v/w.
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33. 5/25/2011
Apparatus used for determination of
Volatile Oil
Fig. 1 33

34.  Determination of Tannins
 Tannins are substances capable of turning animal hides into leather by
binding proteins to form water-insoluble substances that are resistant to
proteolytic enzymes. This process, when applied to living tissue, is
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known as an "astringent" action and is the reason for the therapeutic
application of tannins.
 Chemically, tannins are complex substances; they usually occur as
mixtures of polyphenols that are difficult to separate and crystallize.
 They are easily oxidized, polymerized in solution, form colloidal solution
with water & non crystalline compound.
 Types of tannins:
a) Hydrolysable : Eg. Gallic acid, pyrogallol, ellagic acid etc.
b) Condensed : Eg. Catechutannic acid, catechin etc.
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35. o Procedure :
 Weighed accurately a quantity of powder into a conical flask. Add 150 ml of
water and heat over a boiling water-bath for 30 minutes. Cool, transfer the
mixture to a 250-ml volumetric flask and dilute to volume with water. Allow
the solid material to settle and filter the liquid through a filter-paper,
discarding the first 50 ml of the filtrate.
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 To determine the total amount of material that is extractable into water,
evaporate 50.0 ml of the plant material extract to dryness, dry the residue in
an oven at 105°C for 4 hours and weigh (W1).
 To determine the amount of plant material not bound to hide powder that is
extractable into water, take 80.0ml of the plant material extract, add 6.0g of
hide powder R and shake well for 60 minutes. Filter and evaporate 50.0ml of
the clear filtrate to dryness. Dry the residue in an oven at 105°C and weigh
(W2).
 To determine the solubility of hide powder, take 6.0 g of hide powder R, add
80.0ml of water and shake well for 60 minutes. Filter and evaporate 50.0ml
of the clear filtrate to dryness. Dry the residue in an oven at 105°C and weigh
(W3).
 Calculate the quantity of tannins as a percentage using the following
formula:
W1 - ( W2 - W3 ) / w * 500
 where w = the weight of the plant material in grams.
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36.  Determination of foaming index
 Many medicinal plant materials contain saponins that can cause a
persistent foam when an aqueous decoction is shaken.
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 The foaming ability of an aqueous decoction of plant materials and
their extracts is measured in terms of a foaming index.
o Recommended Procedure:
Weigh accurately 1 gm coarse powder, transferred to 5ooml conical
flask containing 100ml boiling water.
Boil for 30 min
Cool, filter to 100 ml volumetric flask & make up volume to 100ml.
36

37. Take 10 T. T., Mark with 1 to 10, add 1ml, 2ml, 3ml etc. solution to
T. T.
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Make up volume with water to 10 ml in each T. T.
Shake them in a lengthwise motion for 15 seconds, two shakes per
second. Allow to stand for 15 minutes and measure the height of the
foam.
 Result given as :
a) If height of foam is less than 1cm, foaming index is less than 1000.
b) If height of foam is more than 1cm, foaming index is more than 1000.
c) If a height of foam is 1 cm in any tube, then according to volume of
filterate added in tube is used to determine index.
d) Foaming index is calculated by formula : 1000 / a
Where, a = volume of filterate added to test tube
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38.  Determination of Extractable matter
 Determines the amount of active constituents extracted with solvents
from a given amount of medicinal plant material and herbal formulation.
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 Extractive value of crude drug are useful for their evaluation, when the
constituents of a drug can not be readily estimated by any other means.
o Recommended procedure:
Method 1 : Hot extraction
Take accurately weighed coarse powdered air dried material + 100 ml
solvent in a glass stoppered conical flask.
Shake, stand for 1 hr
Boil for 1 hr, cool. Adjust to original volume. Shake & filter.
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39. transfer 25 ml of filterate to a tarred flat bottomed dish and evaporate to
dryness at 105 C for 6 hr & weigh.
Calculate content of extractable matter in mg per gm of air dried material.
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Method 2 : Cold Maceration :
Take accurately weighed coarse powdered air dried material + 100 ml
solvent in a glass stoppered conical flask.
Macerate for 6 hr
Allow to stand for 18 hr, Filter.
transfer 25 ml of filterate to a tarred flat bottomed dish and evaporate to
dryness at 105 C for 6 hr & weigh.
39
Calculate content of extractable matter in mg per gm of air dried material.

40.  Determination of Haemolytic activity:
 Many medicinal plant material, especially those derive from families
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Carryophyllaceae, Araliaceae, Primulaceae and Dioscoreaceae contains
saponins. The most characteristic property of saponin is ability to cause
haemolysis.
 The haemolytic activity of plant material is determined by comparison
with reference material, saponin R, which has haemolytic activity of
1000 units per gm.
 A suspension of erythrocyte is mixed with equal volume of serial
dilution of plant material extract.
 The lowest concentration cause complete haemolysis is determined after
allowing mixtures to stand for a given period of time. A similar test is
carried out simultaneously with saponin R.
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41. o Method:
a) Preliminary test:
 Prepare serial dilution of plant material extract with phosphate buffer
7.4 and blood suspension (2%) using four Test tube as follow:
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Tube No.
1 2 3 4
Plant material extract (ml) 0.1 0.2 0.5 1
Phosphate buffer ph 7.4 0.9 0.8 0.5 -
Blood suspension (ml) 1 1 1 1
 Mix, avoiding formation of foam, allow to stand for 6 hr at room
temperature. Examine tube & record dilution for total haemolysis &
proceed as follows :
a) If total haemolysis is observed only in tube no. 4, use original extract.
b) If observed in 3, 4, prepare 2 fold dilution of original extract.
c) If observed in 2, 3, 4, prepare 5 fold dilution of original extract.
d) If all tubes contain, clear, red solution, prepare 10 fold dilution of 41
original extract.

42. b) Main test:
 Prepare serial dilution of plant material extract diluted / undiluted with phosphate
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buffer 7.4 and blood suspension (2%) using 13 Test tube as follow:
Tube no.
1 2 3 4 5 6 7 8 9 10 11 12 13
Plant 0.4 0.45 0.5 0.55 0.6 0.65 0.7 0.75 0.8 0.85 0.9 0.95 1
material
extract
Phosphate 0.6 0.55 0.5 0.45 0.4 0.35 0.3 0.25 0.2 0.15 0.1 0.05 -
buffer pH 7.4
Blood 1 1 1 1 1 1 1 1 1 1 1 1 1
suspension
 Mix, observe result after 24 hr. Calculate amount material in gm that produce
complete haemolysis.
 Prepare similar dilutions of saponin R & calculate quantity of saponin R in gm that
produce complete haemolysis.
 Calculate haemolytic activity of material using following formula:
1000 * a / b
where, 1000 = defined haemolytic activity of saponin R on ox blood,
a = Quantity of saponin R that produce total haemolysis,
42
b = Quantity of material that produce total haemolysis.

43.  Determination of Radio active contamination:
 There are many sources of ionization radiation, including radionuclides,
occurring in the environment. Hence a certain degree of exposure is
inevitable. Dangerous contamination, however, may be the consequence of a
nuclear accident.
 The WHO, in close cooperation with several other international
organizations (IAEA), has developed guidelines in the event of a widespread
contamination by radionuclides resulting from major nuclear accidents.
These publications emphasize that the health risk, in general, due to
radioactive contamination from naturally occurring radio nuclides is not a
real concern, but those arising from major nuclear accidents such as the
nuclear accident in Chernobyl, may be serious and depend on the specific
radionuclide, the level of contamination, and the quantity of the
contaminant consumed.
 Taking into account the quantity of herbal medicine normally consumed by
an individual, they are unlikely to be a health risk. Therefore, at present,
43
no limits are proposed for radioactive contamination.

44.  List of Q. C. methods for medicinal plant material as
per W. H. O.
 Determination of foreign matter
 Macroscopic and microscopic examination
 Thin-layer chromatography
 Determination of ash
 Determination of extractable matter
 Determination of water and volatile matter
 Determination of volatile oils
 Determination of bitterness value
 Determination of haemolytic activity
 Determination of tannins
 Determination of swelling index
 Determination of foaming index
 Determination of pesticide residues
 Determination of arsenic and heavy metals
 Determination of microorganisms 44
 Radioactive contamination

45.  References:
1. Quality control methods for medicinal plant material (WHO).
2. “Quality standard of indian medicinal plants” ,Indian council of medicinal
research , new delhi-2003 , volume-I.
3. The ayurvedic pharmacopoiea for india, volume-I.
4. “Quality control of herbal drugs-an approach to evaluation of botanicals ” ,
mukherjee p.k. , bussiness horizons pharmaceutical publishers , new
delhi-2002.
5. Indian pharamacopoiea 2007, volume – 3.
6. Pharamacognosy by C. K. Kokate 36th edition, Nirali prakashan.
7. Experimental pharmacognosy by Biren N. Shah.
45
8. www.wikipedia.com

46.  Questions:-
1. Describe quality control methods for medicinal plant material as per
WHO.
2. Enlist quality control methods for medicinal plant material as per WHO.
Describe determination of –
a) Pesticide residue in plant material,
b) Extractable matter,
c) Ash value.
3. Enlist quality control methods for medicinal plant material as per WHO.
How extractable matter from plant material is determined?
4. Define compendial method. Enlist standardisation parameters for
evaluation of herbal drugs. Add a note on determination of Ash value.
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