Chemical tests

Analytical Pharmacognosy
Drug Adulteration
Deterioration- impairment in the quality of drug.

Admixture-addition of one article to another due to carelessness or by accident.

Sophistication- Intentional or deliberate type of adulteration.

Substitution- totally different substance is added in place of original drugs.

Inferiority- adding any substandard drug.

Spoilage- due to attack of microorganisms.

Examples

Substitution with substandard commercial varieties-

1. Presence of Strychnousnux-blanda or S. potatorum in place of S. nux vomica.

2. Capsicum minimum replaced by C. annuum.
3. Indian senna substituted with Arabian senna and dog senna.
4. Gentian substituted by Kutki.
5. Medicinal ginger replaced by its inferior varieties such as African, Japanese and Cochin ginger.

Substitution with superficially similar inferior drugs

1. Belladonna leaves are substituted with Ailanthus leaves.

2. Saffron [Crocus sativus (Iridaceae)] is mixed with dried flowers of Carthamustinctorius.
3. Scented bdellium is used for Myrrh.
4. Mother cloves and clove stalks are mixed with clove [Cloves are the dried flower buds of
Syzygiumaromaticum (Eugenia caryophyllus) (Myrtaceae)]
5. Bees wax is substituted by Japan wax.

Substitution with artificially manufactured substances

1. Compresses chicory in place of coffee.

2. Paraffin wax substituted for beeswax.
3. Properly cut and shaped basswood for nutmeg

Substitution with exhausted drugs

1. Exhausted gentian made bitter with aloes

2. Artificial colouring of exhausted saffron.
3. Addition of synthetic benzyl benzoate to balsam of Peru
4. This type adulteration is more common in volatile oils containing drugs like fennel, coriander,
clove, caraway etc.

Presence of vegetative matter from the same plant

1. The lower plants like moss, liver worts and epiphytes growing on bark portion are mixed with
cascara or cinchona
2. The stem portions are mixed along with leaf drugs like stramonium, lobelia and senna.
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Harmful adulterents
1. Pieces of amber coloured glass in colophony
2. Limestones in asafoetida
3. Lead shot in opium
4. White oil in coconut oil
5. Stearin or paraffin in cocoa butter

Adulteration of Powders

1. Dextrin in ipecacuanha
2. Powdered liquorice or gentian admixed with powdered olive stones.
3. Red sanders wood in capscicum
4. Brick powder in powdered bark.

Famous drugs and adulterants

Drug Adulterants
Digitalis Primrose, Comfrey leaves
Belladona Leaves of Phytolacca Americana, Solanum
nigrum, Ailanthus glandulosa
Senna Tinnaveli Senna (Cassia angustifolia) Dog Senna (Cassia obovate), Palthesenna
(Cassia auriculate), Bombay senna
Alexandrian senna (Cassia acutifolia) Dog senna, Palthesenna, Bombay senna
Clove (flower buds) Mother Clove(ripened fruit), Blown Clove
(expanded flowers), exhausted cloves(it
may float on water), clove stalks [Presence
of sclereids or calcium oxalate crystals in
clove stalk. (genuine powdered clove do not
contain this)].
,

DRUG EVALUATION
1. Morphological or Organoleptic Evaluation
Organoleptic evaluation means conclusions drawn from studies resulted due to impressions on
organs of senses.
Morphology- The study of form of a crude drug
Morphography- Description of the form of the drug.
Examples:
1. Fractured surfaces in cinchona, quillaia and cascara barks and quassia wood
2. Aromatic odour of umbelliferous fruits and sweet taste of liquorice
3. Ovoid tears of gum acacia, ribbon shaped characteristics of tragacanth, disc shaped structure
of nux-vomica, conical shape of aconite, quills of cinnamon
4. The wavy shape of rauwolfia, pungent taste of capsicum, brown colour of cinnamon, odour
and taste of spices
2. Microscopic Evaluation
Detailed examination of organised drugs by their known histological characters.
Examples:
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Microchemistry
1. A drop of phloroglucinol and Conc. HCl give pink stain with lignin
2. Mucilage is stained pink with rhuthenium red
3. Cellulose swells and dissolves in cuoxam (Ammoniacal solution of Copper oxide)(used as test
for cotton)
4. Hemicellulose and starch stained blue with N/50 iodine solution

Microanatomy

5. Identification of Lignified trichomes in nux vomica

6. Warty trichomes of senna
7. Wavy medullary rays of cascara bark
8. Glandular trichomes of mint
9. Presence of sclereids or calcium oxalate crystals in clove stalk. (genuine powdered clove do not
contain this).
10. Powdered clove fruits shows presence of starch, while it is absent in clove
11. Presence of non lignified vessels in powders of rhubarb and ginger indicates adulteration
12. Senna varieties are distinguished by stomatal number and palisade ratio.
13. The diameter of starch grains in Cinnamomum cassia is 10 microns and hence, useful for
detecting adulterants.
14. The number of sclerenchymatous cells/mm2 of cardamom is useful for detecting different
varieties of cardamom seed.
Leaf constants:
 Palisade ratio-Average number of palisade cells beneath each epidermal cell.it can
be determined with powdered drugs by using camera lucida
 Atropa belladonna 6 - 10
 Digitalis lanata 2.5 - 6.5
 Cassia angustifolia 5.5 – 10
 Vein-islet number- Number of vein-islets/mm2of the leaf surface midway between
the midrib and margin.

 Vein-termination number- the number of veinlet terminations per mm2 of leaf

surface. A vein termination is the ultimate free termination of a veinlet or branch
of a veinlet. By this character it is possible to distinguish between Peruvian and
Bolivian coca Ieaves and between Alexandrian and Tinnevellysenna leaflets.
 Stomatal number-Average number of stomata/mm2 of epidermis of the leaf
Name of drug Stomatal number
Upper lower
Atropa belladonna 5 – 14 78 – 95
Cannabis sativa - 423-550
Centella asiatica 66-82 141-228
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Cassia angustifolia 220-260 240-265

Mentha piperita 30-65 325-375
Ocimum sanctum 64-72 175-250

 Stomatal index-the percentage which the number of stomata form to the total
number of epidermal cells; each stoma being counted as one cell.
𝑆
S.I= 𝐸+𝑆*100
S- Number of stomata per unit area
E-Number of epidermal cell in the same unit area.

Stomata Vein termination

Name of drug Stomatal index

upper lower
Atropaacuminata 2.5-4.7 15.5-17.9
Azadrachtaindica - 8-8.5
Eucalyptus globules 4-5 3.5-4.2
Ocimum sanctum 8-8.5 13.1-16.6

Calcium oxalate crystals types

Calcium oxalate crystals by virtue of its peculiar shapes can be utilized for crude drug evaluation

 Cubic (cube shape) e.g,senna, Glycyrrhiza.

 Tetragonal e.g,onion.
 Mono clinic(all three axes are un equal) e.g. Gall.
 Rosettes -clusters (aggregation of crystals) e.g rhubarb, stramonium, cascara, senna, clove, jalap
 microsphenoidal or sandy crystals (belladonna)
 prisms (senna, hyoscyamus, quassia, liquorice, cascara, quillaia, rauwolfia, calumba);
 single acicular crystals (ipecacuanha, gentian, cinnamon);
 bundles of acicular crystals (squill);
 Rhombic (diamond shape)

Trichomes(Plant Hairs):

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Trichomes are the tubular elongated or glandular outgrowth of the epidermal cell.
Trichomes consists of two parts viz., root(in the epidermis) and body (outside the epidermis).
Trichomes may be present in various aerial plant part such as leaves (senna and digitalis), seeds
(nux vomica and strophanthus), fruits (ladies finger) etc. (trichomes are absent in roots).
Trichomes are as such as functionless, but sometimes, perform secretory functions (secrete water
in most case, volatile oil as in case of peppermint)

Functions of Trichomes:
1. Generally a dense covering of woolly trichomes controls the rate of transpiration.
2. They aid in the protection of plant body from outer injurious agencies.

Depending upon the structure and number of cells present in trichomes, they are classified as:

1. Covering trichomes or Non glandular trichomes or clothing trichomes

2. Glandular Trichomes
3. Hydathodes or special types of trichomes

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I. COVERING OR NON-GLANDULAR OR CLOTHING TRICHOMES

a. Unicellular:
1. Liginified trichomes: Nux vomica, strophanthus
2. Short, sharoly pointed, curved: Cannabis
3. Large, conical, strongly shrunken: Lobelia
4. Short, conical, unicellular: Tea, buchu
5. Strongly waved, thick walled: Yerba Santa
b. Multi Cellular (Pluricellular) –unbranched trichomes
i. Uniseraite (single raw of cells)
1. Bi cellular, Conical: Datura
2. Three celled long: Stramonium
3. Three to four celled long: Digitalis
4. Four to five celled long: Belladonna
ii. Biseriate (Multi raws of cells): Calendula officianalis
iii. Multiseriate: Fern
c. Multi cellular branched trichomes:
1. Stellate: Hammamelis, Helicteris-isora
2. Peltate: (plate like arrangement of surrounded cells): Humulus
3. Candelabra (Uniseriate branched axis): Verbascum thapsus
4. T Shaped trichomes: Artemisia, pyrethrum
II. GLANDULAR TRICHOMES
These are characterised by the presence of glandular (spherical) cell all the top of the
trichome.These are sub-classified as under:
1. Unicellular glandular trichomes: The stalk is absent eg. Piper. Betel Vasaka
2. Multicellular glandular trichomes
a. Trichomes with unicellular head and unicellular stalk, eg Digitalis purpurea.
b. Unicellular head and uniseriate multicellular stalk, eg Digitalis thapsi, belladonna,
etc
c. Multicellular head, multicellular, biseriate stalk, eg: Santonica and plants of
Compositae, such as sunflower, etc.
d. Unicellular stalk and biseriate head, eg. digitalis purpurea.
e. Short stalk with secreting head formed of rosette or club shaped cells, eg. Mentha
species
f. Trichomes with multicellular, multiseriate. Cylindrical stalk and a rosette of
secretory cells. Eg. Cannabis sativa
g. Multicellular multiseriate head and multicellular uniseriate stalk, eg Indian hemp
and tobacco
III. HYDATHODE (Special types of trichomes)
These are organs of absorption or secretion of water developed in certain plants eg:
Piper betal, London pride etc.

STOMATA
 It is a tiny opening or pore that is used for gas exchange.
 They are mostly found on the under-surface of plant leaves.
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 In a stoma, there is the chloroplast, a cell wall, a vacuole and a cell nucleus.
 In aquatic plants stomata is absent.
Types of stomata
1.Moss Type
2.Gymnospermous Type
3.Gramineous Type
4.Dicotyledonous Type

 Dicotyledonous stomata classified as

Actinocytic (meaning star-celled)

 stomata have guard cells that are surrounded by at least five radiating cells forming a star-
like circle. This is a rare type that can for instance be found in the Ebenaceae family.
Anisocytic (meaning unequal celled)
 stomata have guard cells between two larger subsidiary cells and one distinctly smaller
one. This type of stomata can be found in more than thirty dicot families, including
Brassicaceae, Solanaceae, and Crassulaceae. It is sometimes called cruciferous type.
Anomocytic (meaning irregular celled)

 stomata have guard cells that are surrounded by cells that have the same size,
shape and arrangement as the rest of the epidermis cells. This type of stomata can
be found in more than hundred dicot families such as Apocynaceae, Boraginaceae,
Chenopodiaceae, and Cucurbitaceae. It is sometimes called ranunculaceous type.
Diacytic (meaning cross-celled)

 stomata have guard cells surrounded by two subsidiary cells, that each encircle one
end of the opening and contact each other opposite to the middle of the opening.
This type of stomata can be found in more than ten dicot families such as
Caryophyllaceae and Acanthaceae. It is sometimes called caryophyllaceous type.
hemiparacytic

 stomata are bordered by just one subsidiary cell that differs from the surrounding
epidermis cells, its length parallel to the stoma opening. This type occurs for
instance in the Molluginaceae and Aizoaceae.
Paracyticor Rubiaceous(meaning parallel celled)

 Stomata have one or more subsidiary cells parallel to the opening between the
guard cells. These subsidiary cells may reach beyond the guard cells or not. This
type of stomata can be found in more than hundred dicot families such as
Rubiaceae, Convolvulaceae and Fabaceae. It is sometimes called rubiaceous type.

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
Types of stomata example

Diacytic (Caryophyllaceous or cross Vasaka

celled stomata) PeppermintSpearmint
Anisocytic (cruciferous or unequal celled Belladonna
stomata) Datura,Stramonium
Anomocytic(Ranunculaceous or irregular Digitalis
celled stomata) Lobelia
Buchu
Paracytic or rubiaceous or parallel celled Coca leaf, Senna leaf
stomata

QUANTITATIVE MICROSCOPY
Lycopodium spore method
 Method to determine the percentage purity
 Percentage purity of drug = N*W*94000*100
S*M*P
n=no.of characteristic structures
w=weight in mg of lycopodium taken
s=no.of lycopodium spores
M=weight in mg of the sample
P= 2,86,000 in case of ginger starch grains

Chemical evaluation
The chemical evaluation includes qualitative

 Chemical tests,
 Quantitative chemical tests,
 Chemical assays and Instrumental analysis.
The isolation, purification and identification of active constituents are chemical methods of
evaluation. Qualitative chemical tests include identification tests for various phytoconstituents
like alkaloids, glycosides, tannins, etc.
Eg.

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 Copper acetate used in the detection of colophony present as an adulterant.

 Van Urk’s reagent for ergot
 Vitali morins reaction for tropane alkaloids
 Iodine for starch
Quantitative chemical tests such as acid value(resins, balsams), saponification value(balsams),
ester value (balsams, volatile oils), acetyl value (volatile oils), etc. are also useful in evaluation of a
drug by means of chemical treatment.
Chemical assay include assays for alkaloid, resin, volatile oils, glycoside, vitamins or other
constituent.
Eg.

 Assay of total alkaloid in belladonna herb,

 The total alkaloid and nonphenolic alkaloid in ipecacuanha,
 The alkaloid strychnine in nux vomica,
 The resin in jalap and
 The vitamins in cod -liver oil.
The results obtained can conclude the presence of inferior or exhausted drug and, by proving
absence of the assayed constituent.
Instrumental analyses are used to analyse the chemical groups of phytoconstituents using
chromatographic spectroscopic methods.
CHEMICAL TESTS FOR DETECTION OF PLANT CONSTITUENTS

Chemical Tests / Reagents Inferences

constituents
Dragendorff’s reagent: Potassium bismuth iodide solution Reddish brown ppt

Mayer Test –Potassium Mercuric iodide Cream color ppt

ALKALOIDS
(Common test) Hager Test – Picric Acid (saturated) Yellow ppt

Wagner Test- Iodine in KI solution Reddish brown ppt

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Ergot alkaloids Van urk’s reagent:

Dissolve 1 g of p-dimethyl-amino benzaldehyde in 50 ml of Blue colour
hydrochloric acid and 50 ml of 95 % ethanol.

Purine Murexide test

alkaloids Potassiumchlorate+HCl……>Exposingtheresidue Purple colour
to ammoniavapour

Tropane Vitali- morin reaction

alkaloids Fuming HNO3 + dry to get residue + add Methanolic Violetcolour
potassium hydroxide to an acetone solution of nitrated
residue

Cinchona Thalleioquin test

alkaloids Bromine water + ammonia solution Green colour
(Quinine &
quinidine
alkaloids)

Ipecacuanha Frohde’s Test Bluish green colour

alkaloids Sodium molybdate + HCl

Baljet test- Picric acid + NaOH Appearance of orange

or red colour

Legal test – Pyridine + sodium nitroprusside + NaOH Pink or deep red

colour

Cardiac Keller- killiani test : (For deoxy sugars) Glacial acetic acid Appearance of a pink
glycosides +FeCl3 + H2SO4 or deep red colour

Xanthydrol test: Xanthydrol + Glacial acetic acid+ HCl Red colour

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Antimony trichloride test :(cardenolides &Bufadienolides ) Blue or violet colour

Antimony trichloride + Trichloroacetic acid

Keddes test: (For cardenolides) blue or violet colour

3, 5 dinitrobenzoic acid in methanol + KOH soln that fades out in 1 to
2 hours
Libermann Burchard Test:
To the solution of glycosides is added in acetic Violet to blue colour
anhydride followed by concentrated sulphuric acid.

Raymond test
glycosides + 50%ethanol + 1% dinitro benzene in ethanol or Appearance of violet
methanol + 2-3 drops of 20% NaOH sol is added. color,which changes
into blue colour
(Or)

Test solution+hot methanolic alkali. Violet colour is

produced

Saponin Foam test: The powdered drug is shaked well with water. Foam produced
glycosides

Anthraquinone Borntrager Test Pink, red or violet

glycosides Drug+ Benzene /Chloroform — separate the organic colour
solvent and then add ammonia

Modified Borntrager Test rose-pink to cherry

(For C-Glycosides) red colour
FeCl3 + Dil. HCl + Benzene /CCl4 — separate the organic
solvent and then add ammonia

Flavanoid Shinoda test : 95% ethanol + Con. HCl + Mg Pink colour

glycosides

Fehling’s solution (For reducing sugar) Brick red ppt

copper sulphate + water (solution A). potassium sodium
Carbohydrate tartrate + sodium hydroxide (solution B).
Mix two solutions in equal volume prior to use

Iodine water (For starch) starch grains show

(potassium iodide in 100 ml of water) blue color

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Molisch test Purple colour

a-napthol + H2SO4

Test for mucilage Red colour

Ruthenium red

Osazone formation test

Phenyl hydrazine hydrochloride + sodium acetate+ acetic Yellow colour
acid

Resorcinol test /Selivanoff’s test ( for ketones) Rose colour

Resorcinol crystal + HCl

Killer-kilani test for deoxy sugars:

A deoxy sugar is dissolved in acetic acid containing a trace A reddish-brown color
of FeCl3 and transferred to the surface of concentrated is formed at the
H2SO4 junction which
turns blue

Benedict’s test:
To the solution, add benedicts reagent and heated on water Solution appears
bath green,yellow or red

Phenols Ferric chloride(Alcoholic) blue color

A 5% w/v solution of ferric chloride in 90 % alcohol

Millon’s test Protein is stained red

mercury + nitric acid
Proteins
Biuret Test (General test) – Violet or pink colour
NaOH + 1 % CuSO4

Lead sulphide test:

To the alkaline solution of sulphur containing proteins add Black ppt
lead acetate

Heat coagulation test: Proteins get ppt

Heat the test solution in a boiling water bath

Amino acids Ninhydrin reagent Pink ,blue or violet

0.1% solution in butanol

Fats & oils Sudan red III Red coloiur

Sudan red in 20 ml of alcohol (90 %) and 20 ml of glycerin

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Halphen’s test/bevan’s test:

Fixed oils oil + amyl alcohol+ 1% solution of sulphur in CS2 for 10 Red colour
minutes in a water bath Indicate presence of
cotton seed oil

Boudouin’s test:
The oil is shaken with half its volume of con. HCl containing Pink colour
1% of sucrose Presence of seasame
oil

Phloroglucinol solution It stains lignified tissue

Dissolve 2 g of phloroglucin in 100 ml of 90 % alcohol to pinkish red
Lignified cells Alongwith conc, hydrochloric acid (1:1)

Safranin It stains lignified cells

It is 1% safranin solution in 50 % ethyl alcohol red
Goldbeater’s skin test:
A small piece of goldbeater’s skin is soaked in2% HCl +water
and placed in a solution of tannin for 5min.

The skin piece is washed with distilled water and kept in A brown or black
a solution of FeSO4. colour on skin

Tannins Vanillin-Hydrochoric Acid Test:

When the drug is treated with Vanillin-Hydrochoric Acid Pink or red colour
reagent

Gelatin Test:
To a sol of tannin (0.5-1%) aquous sol of gelatin (1%) and White buff coloured
NaCl (10%) are added. ppt

Catechin test (matchstick test):

A matchstick is dipped in aq plant extract, dried near burner On warming near a
and moistened with conc HCl flame the matchstick
wood turns pink or
red due to formation
of phlorogucinol.
Gambir-flurescin test:
A mixture of alcoholic extract of pale catechu (1g)NaOH Sol The petroleum ether
(5ml) nd petroleum ether (5ml) is shaken and kept for layer shows green
sometime. fluorescence

Physical evaluation
Moisture content, specific gravity, density, optical rotation, refractive index, melting point,
viscosity, and solubility in different solvents.
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Analytical Pharmacognosy

1. Moisture content- The moisture content of a drug will be responsible for

decomposition of crude drugs either producing chemical change or microbial growth.
So, the moisture content of a drug should be determined and controlled. The
moisture content is determined by heating a drug at 105o c in an oven to a constant
weight.

Eg. The moisture content of digitalis should not be more than 6%W/W
2. Solubility: Drug specific behaviour towards solvents are taken into consideration.
Eg. Solubility of colophony of colophony in light petroleum, the solubility of balsam of Peru
in solution of chloral hydrate
3. Optical rotation :Anisotropic crystalline solids and samples containing an excess of
one enantiomer of a chiral molecule can rotate the orientation of plane polarized
light. Such substances are said to be optically active, and this property is known as
optical rotation.
Eg. Eucalyptus oil (0o to +10o ),
honey (+3o to -15o )
4. Refractive index: It is defined as the property of a material that changes the speed of
light, computed as the ratio of the speed of light in a vacuum to the speed of light
through the material. when light travels at an angle between two different materials,
their refractive indices determine the angle of transmission refraction of the light
beam. This could be used as a parameter in evaluating the herbal drugs.
Eg castor oil 1.4758-1.527
5. Specific gravity: It is also known as relative density. The ratio of the mass of a solid or
liquid to the mass of an equal volume of distilled water at 4 o c(39o F) or of a gas to an
equal volume of air or hydrogen under prescribed conditions of temperature and
pressure.
Eg. Specific gravity of drugs are cottonseed oil 0.88-0.93, coconut oil 0.925, castor oil
o.95,etc.
6. Viscosity: Viscosity of a liquid is constant at a given temperature and is an index of its
composition. Eg.pyroxylin kinematic viscosity, 1100-2450 centistokes.
7. Melting point: Plant constituents have very sharp and constant melting points. As far
as crude drugs are concerned, melting point range has been fixed due to the mixed
chemicals. Eg. Beeswax 62-65o c,wool fat 34-44o c
8. Ultraviolet light: Certain drugs fluoresce when the cut surface or the powder is
exposed to ultraviolet radiation, and it is useful in the identification of those drugs.
Eg. Some pieces of rhapontic, Indian and Chinese rhubarb are very difficult to distinguish,
and it is very difficult in powdered form, but examination in ultraviolet light gives such
marked differences in fluorescence that the varieties can be easily distinguished from each
other.
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9. Ash value: The determination of ash is useful for detecting low grade products,
exhausted drugs, and excess of sandy or earthy matter. Different types of ash values
are used in detection of crude drugs are,
 total ash
 acid insoluble ash
 water- soluble ash
 sulphated ash.
Total ash is useful in detecting the crude drugs that are mixed with various mineral
substances like sand, soil, calcium oxalate, chalk powder or other drugs with different
inorganic contents to improve their appearance, as is done for nutmegs and ginger.
Acid insoluble ash means the ash insoluble in dilute hydrochloric acid. The majority of
crude drugs contain calcium oxalate, and the quantity of calcium oxalate varies very
frequently.
Eg. Rhubarb, total ash range from 8-40%. In this case, the total ash is useless to detect
earthy matter adherent to such a drug. So, acid insoluble ash would be preferable for
rhubarb.
10. Extractive values: The extracts obtained by exhausting crude drugs with different
solvents are approximate measures of their chemical constituents. Various solvents
are used according to the type of the constituents to be analysed.
Water soluble extractive is used for crude drugs containing water-soluble
constituents like glycosides, tannins, mucilage etc;
alcohol- soluble extractive is used for crude drugs containing tannins, glycosides,
resins, etc; and
ether-soluble extractives are used for drugs containing volatile constituents and fats.
11. Foreign organic Matters- The parts of the organ or organs other than those parts of
drugs mentioned in the definition and description of the drug are known as foreign
organic matters. They may be insect, moulds, earthy material, animal excreta, etc.
Eg. Garlic should not contain more than 2%,

saffron should not contain more than 2%.

Biological evaluation: When the estimation of potency of crude drug or its preparation
is done by means of its effect on living organisms like bacteria, fungal growth or animal
tissue or entire animal, it is known as bioassay. This method is generally called for, when
standardisation is not adequately done by chemical or physical means and also for
conformity of therapeutic activity of raw material and finished product. In other words,
bioassay is the measure of sample being tested capable of producing biological effect as
that of the standard preparation. Such activity is represented in units known as
international unit (I.U.) The specific biological activity contained in each I.U. of the few drugs
is mentioned as under:
Digitalis- 1IU is contained in 76 mg of standard preparation
VitA- 1 IU is present in 0.344 micrograms of standard preparation

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Analytical Pharmacognosy

Biological assy methods are mainly of 3 types

 Toxic
 Symptomatic
 Tissue methods
In toxic and symptomatic techniques, the animals (like rabbit, guinea pig, pigeon etc) are
used, whereas in tissue method, the effect of a drug is observed on isolated organ or tissue.
Among the drugs that are subjected to bioassay are cardiac glycosides (pigeon is the testing
animal), natural pesticides and antibiotics.

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