Adulteration and Evaluation of

crude drugs
 Adulteration

 Adulteration is defined as substituting the original crude drug partially or wholly with
other similar looking substances which are free from or inferior in chemical and
therapeutic property.
 Adulteration involves
 Deterioration- Impairment in quality of drug
 Admixture- Addition of one article to another due to ignorance/ carelessness or by
accident.
 Sophistication: Intentional/ deliberate addition of unwanted material
 Substitution: Addition of totally different substance instead of the original.
 Inferiority: Substandard drug
 Spoilage: due to microorganisms
 Types of Adulteration

 Unintentional adulteration
 Intentional adulteration

Unintentional adulteration may be due to the following reasons

 Confusion in Vernacular names
 Lack of knowledge about authentic source
 Similarity in morphology
 Lack of authentic plant
 Similarity in Color
 Careless Collections
 Unintentional adulteration

 a. Confusion in Vernacular names

Same vernacular name of different species and different vernacular

names of same species creates confusion and invites adulteration. In

Ayurveda, Parpatta refers to Fumaria parviflora. In Siddha,

“Parpadagam” refers to Mollugo pentaphylla. Owing to the similarity in

the names in traditional systems of medicine, these two herbs are often

interchanged or adulterated or substituted

Language Vernacular name Scientific name Family
Same vernacular name of different species
Dissotis rotundifolia Melastomataceae
(Smith) Triana
Kibombo
Rauvolfia Apocynaceae
mombasiana Stapf.
Digo
Kamata Aerangis thomsonii Orchidaceae
(Rolfe) Schltr.
Culcasia scadens Araceae
P.Beauv

Different vernacular names of same species

Mnkande; Kigelia africana Bignoniaceae

Shambaa Mvugve; (Lam.) Beneth.
Mvungunya
b. Lack of knowledge about authentic source :
Nagakesar is one of the important drugs in Ayurveda. The authentic source is Mesua ferrea.
However, market samples are adulterated with flowers of Calophyllum inophyllum because
suppliers are unaware of it. Authentic flowers can be easily identified by the presence of two-
celled ovary whereas in case of spurious flowers they are single celled.
c. Similarity in morphology
Mucuna pruriens is adulterated with other similar Papilionaceae seeds having similarity in
morphology. Mucuna utilis (sold as white variety) and Mucuna deeringiana (sold as bigger
variety) are popular adulterants. Apart from this Mucuna cochinensis, Canavalia virosa and
Canavalia ensiformis are also sold in Indian markets. Authentic seeds are up to 1 cm in length
with shining mosaic pattern of black and brown color on their surface. Mucuna deeringiana and
Mucuna utilis are bigger (1.5-2 cm) in size. While Mucuna deeringiana is dull black and
Mucuna utilis is white or buff colored.
d. Lack of authentic plant
Hypericum perforatum is cultivated and sold in European markets. In India, availability of this
species is very limited. However, the abundant Indo-Nepal species Hypericum patulum, sold in
the name of Hypericum perforatum. Market sample is a whole plant with flowers and it is easy to
identify them taxonomically. Anatomically, transverse section of Hypericum perforatum stem has
compressed thin phloem, hollow pith and absence of calcium oxalate crystals. Whereas
Hypericum patulum as broader phloem, partially hollow pith and presence of calcium oxalate
crystals.
e. Similarity in Color
With course of time, drug materials get changed to or substituted with other plant species.
„Ratanjot‟ is a recent day example. In the past, roots of Ventilago madraspatana were collected
from Western Ghats, as the only source of „Ratanjot‟. It is clearly known that Arnebia euchroma
var. euchroma is the present source. Similarity is in yielding a red dye, Arnebia euchroma
substitutes Ventilago madraspatana.
f. Careless Collections
Some of the herbal adulterations are due to the carelessness of herbal collectors and
suppliers. Parmelia perlata is used in Ayurveda, Unani and Siddha. It is also used as
grocery. Market samples showed it to be admixed with other species (Parmelia
perforata and Parmelia cirrhata). Sometimes, Usnea sp. is also mixed with them.
Intentional adulteration

a. Substitution with inferior commercial varieties

b. Substitution with artificially manufactured drug
c. Substitution by exhausted drugs
d. Substitution by superficially similar but cheaper natural substances
e. Substitution by addition of worthless or heavy materials
f. Addition of synthetic principles
a. Substitution with inferior commercial varieties

It is the use of morphologically similar, different inferior commercial verities (may or

may not have any chemical or therapeutic potential as that of original natural drug).
Example are
 Arabian senna (Cassia angustifolia), dog senna (Cassia obovata) and avaram senna
(Cassia auriculata) have been used to adulterate Senna (Cassia senna).
 Japanese ginger (Zingiber mioga) have been used to adulterate medicinal ginger
(Zingiber officinale).
 Capsicum annum have been used to adulterate Capsicum minimum.
 Piper nigrum fruit is adulterated by Carica papaya seeds.
b. Substitution with artificially manufactured drug
Artificially manufactured substances use as a substitute of the original drug. Artificial sugar
for honey, yellow colored paraffin wax for bees wax, compressed Chicory in place of
coffee and properly cut and shaved basswood for nutmeg (Jaijifal) are examples.
c. Substitution by exhausted drugs
Same plant material is mixed with drug having no active medicinal components as they
have already been extracted out.
Examples
 Volatile oil containing drugs : Foeniculum vulgare (fruit / fennel) , Syzygium
aromaticum (flowering buds / clove) , Coriandrum sativum (fruit / coriander) , Carum carvi
(fruit / caraway / siahjeera) , Cascara sagrada (Sacred Bark / jamal gota) and Zingiber
officinale (roots / ginger).
Coloring matter containing drugs: In case of loss of coloring material during
exhaustion the residue is recolored with artificial dye. Examples: Rosa macdub (Red rose
petal) and Crocus sativus (stigma of flowers / saffron), Camellia sinensis (leaves / tea).

d. Substitution by superficially similar but cheaper natural substances

Adulterated product has no relation with genuine material, may or may not have any
therapeutic or chemical component. As Ailanthus altissima (Ailanthus) are substituted
for Atropa belladonna (Belladonna), Cassia acutifolia (senna) , Mentha longifolia (mint)
etc.; Leaves of Phytolacca americana (pokeweed) and Scopolia japonica, (Japanese
belladonna) for Atropa belladonna (Belladonna); Leaves of Xanthium strumarium for
stramonium and dandelion Anethum sowa (Indian dill) with Anethum graveolens
(Europian dill) or Carum carvi (caraway).
e. Substitution by addition of worthless or heavy materials
Examples
 Large mass of stone mixed with Glycyrrhiza glabra (liquorice root).
 Pieces of lime stone mixed with Ferula foetida (Asafoetida).
 Lead shot mixed with pieces of Papaver somniferum (opium).

f. Addition of synthetic principles

It is the use of synthetic chemicals to enhance the natural character.
Addition of benzyl benzoate to Peru balsam, citral to citrus oils and lemon oil , orange oil.
EVALUATION OF CRUDE DRUGS

Evaluation of a drug ensures the identity of a drug and

determines the quality & purity of the drugs.

Evaluation of the drug indicates :

 Biochemical variation in the drug
 Effect of treatment and storage of drugs
 Adulteration and Substitution of drugs
 Methods of Drug Evaluation:

 Types of drug evaluation include:

(1) Organoleptic/ macroscopical evaluation
(2) Microscopic evaluation
(3) Biological evaluation
(4) Chemical evaluation
(5) Physical evaluation
 Organoleptic evaluation

 The term organoleptic evaluation refers to the sensory evaluation.

 This refers to drug evaluation by means of our sense organs like
examination of the odour, colour, taste and texture of the drug.
 It is a qualitative Evaluation.
 Examples:
 Colour:- (Cinnamon Bark -Brown)
 Odour:- (Jatamansi-Aromatic)
 Taste:- (Capsicum-Pungent)
 Size:- (Digitalis--10-30 cm long and 4-10 cm wide)
 Shape:- (Nux vomica-Disc shaped)
 Texture:- (Cascara barks- Fractured surface)
 Microscopic Evaluation

 Qualitative Microscopy:
This method is used to identify organized drug by their known
histological characters through Transverse section (T.S.) or
Longitudinal Section(L.S.) or Radial Longitudinal Section (R.L.S.) or
Tangential Longitudinal Section (T.L.S.). Microscopic Evaluations
also covers study of different constituents by using staining Reagents.
Ex:
Lignin stains red or pink with phloroglucinol and concentrated HCl,
Mucilage is stained pink with ruthenium red
Starch is stained blue with N/50 iodine solution.
 The characteristic features of the cellwalls, cell contents, starch grains, calcium oxalte crystals,
trichomes, fibres, vessels etc are studied in detail.
Eg:
Nuxvomica- lignified trichomes and plasmodesmata
Cascara bark- presence of medullary rays and stone cells
Cloves – Absence of sclereids and calcium oxalate crystals
 Linear measurements include measurement of size of starch grains, length and width of fibres,
trichomes.
Eg:
Diametre of strach grains helps in distinguishing ipecacuanha varities, cassia bark from cinnamon.
Diametre of Phloem fibres helps in distinguishing cassia bark from cinnamon
Width of the vessels helps to detect clove stalks from powdered cloves.
 Microscope can also be used for a quantitative evaluation of
drugs and adulterated powders.
 This is done by counting a specific histological feature such
as,
 Stomatal Number
 Stomatal Index
 Vein-islet Number
 Palisade Ratio
 Quantitative Microscopy
 Stomatal Number:
 The average number of stomata present per square millimeter of the epidermis is known
as stomatal number.
 Stomatal number is relatively a constant for a perticular species of same age and hence,
it is taken into consideration as a diagnostic character for identification of a leaf drug.
 e.g.: Atropa belladonna: {6.0 to 14-37.5 (Upper Surface), 62.5 to 93-174 (lower
Surface)}
 Stomatal Index:
 It is the percentage proportion of the number of stomata to the total number of epidermal
cells.
 Stomatal number varies considerably with the age of the leaf but stomatal index is
relatively constant for a given species.
 It is calculated by the formula
S.I. = S*100/E+S
Where, S.I. = Stomatal Index; S= Number of stomata per unit area;
E= Number of Epidermal cells in the same unit area.

 Example: Atropa belladonna:- 2.3-3.9 to 10.5 (Upper Surface), 20.2 to 21.7- 23.0
(Lower Surface)
Vein-islet Number:
The term “vein-islet” is used for the minute area of photosynthetic tissue encircled by the
ultimate divisions of the conducting strands.
Vein-islet number is defined as the number of vein-islets per sq.mm. of leaf surface.
It is constant for a given species of the plant. It is irrespective with the age factor.

Eg: Digitalis Lanata — 2.0-8.0

Digitalis Purpure — 2.0-5.5
Vein Termination Number:
It is defined as average number of Vein terminations per square millimeter of the leaf
surface midway between midrib and the margin.
Eg: Atropa belladonna — 6.3-10.3
Atropa acuminate — 1.4-3.5
Palisade ratio:

It represents the average number of palisade cells beneath one epidermal cell, using four
continuous epidermal cells for the count.
It is determined from powdered drugs with the help of camera lucida.

e.g. Atropa belladonna (6-10), Digitalis lanata (2.5-6.5)

Quantitative Microscopy:

It is an important analytical technique for powdered drug, especially when chemical and
other methods of evaluation of crude drug fail as accurate measure of quality.

Example: Lycopodium- spores are very characteristic in shape and appearance.

Chemical Evaluation:
Chemical nature of the constituents can be used as tool to device a method for the analysis
of the constituents. It involves chemical tests, chemical assay and also the phytochemical
investigation of the crude drugs.

Chemical Assays
Specific assays for different active principles e.g.total alkaloids, glycosides, resins, tannins,
volatile oil, saponins etc. is carried out by different chemical tests.

Chemical Test
Chemical tests are used for determination of specific chemical constituents which may be
present in any drug to which its therapeutic activity is attributed.
Chemical test Chemical group Result
Dragendorff’s Test Alkaloids Yellow orange precipitate

Borntrager’s Test Antharaquinone Pink Ammonical layer

Glycosides
Shinoda Test Flavonoids Pink colour

Ferric Chloride Test Tannins Blue colour

Molisch’s tests Carbohydrates Purple to violet colour

rings

Millon’s tests Amino acids White precipitate

ISOLATION OF PHYTOCONSTITUENTS
 Physical Evaluation
 In this Method Herbal dugs are evaluated on the basis of some
important physical properties .
 A few of them are:-
 Moisture Content
 Viscosity
 Melting point
 Optical Rotation
 Refractive Index
 Ash Content
 Extractive values
Moisture Content:

Presence of moisture in a crude drug can lead to its deterioration due to either
activation of certain enzymes or growth of microbes.

Moisture content can be determined by heating the drug at 150 ⁰C in an oven to a

constant weight and calculating the loss of weight.
Example: Digitalis: NMT 5%, Ergot: NMT 8% W/W

Viscosity:

Viscosity of a liquid is constant at a given temperature and is an index of its

composition.
Hence, it is used as a means of standardising liquid drugs.

Example:
Liquid paraffin – less than 64 centistokes.
Melting Point:
It is one of the parameters to judge the purity of crude drugs containing lipids as
constituents.
They may of animal or plant origin and contain fixed oils, fats and waxes.
The purity of the following crude drugs can be ascertained by determining their
melting points in the range shown against each of them
Example: Coca butter (30⁰ - 33⁰C), bees wax- ((62 ⁰ - 65 ⁰C)

Optical Rotation:
Many substances of biological origin, having a chiral centre, can rotate the plane of
polarised light either to right or to the left.

The extent of rotation is expressed in degrees, plus(+) indicating rotation to the right
and minus(-) indication rotation in the left.

Such compound are optically active and hence called optical rotation.
Example: eucalyptus oil- (0 to +10), honey –(+3to -15)
Refractive Index:
When a ray of light passes from one medium to another medium of different
density, it is bent from its original path.
Thus, the ratio of velocity of light in vaccum to its velocity in the substance is said
to the Refractive index of the second medium.
It is measured by means of refractometer.

Example: Arachis oil - 1.4678-1.4698

 Ash Content:
The residue remaining after incineration of a known quantity of the air dried crude
drug, is known as the ash content of the drug.
Ash simply represents the inorganic salts naturally occuring in drug or adhering to
it or deliberately added to it as a form of adulteration.
Example: Ginger- Total ash -6%
Ginger – water soluble ash-1.7%
Extractive values:
In crude drugs, sometimes the active chemical constitutes cannot be determined
by normal procedures.
In such cases, water, alcohol or ether soluble extractive values are determined
for evaluation of such drugs.

Example: Water soluble extracts like Aloe vera- NLT 25% W/W
Alcohol soluble like Aloe vera- NLT 10 % W/W
Ether soluble extractive valve of linseed- NLT 25% W/W

 Ultraviolet light:
Ultraviolet light is also used for determing the fluorescence of extracts of some
drugs.
Example: Indian and chinese rhubarb
 Biological evaluation

It is employed when the drug cannot be evaluated satisfactorily by chemical and

physical methods.
In this method, the response produced by the test drug on a living system is compared
with that of the standard preparation.
These methods are performed on living animals, isolating living organ and tissue,
animal preparation, and micro-organism ( Bioassay)
Indications of Biological Evaluation:
- When the chemical nature of the drug is not known but is has an biological action.
- When chemical methods are not available.
- When the quantity of the drug is small and so it cannot be evaluated chemically.
- Drugs which have different chemical composition but same biological activity.
BIOASSAY
When the estimation of crude drug or its preparation is done by means of its effect on living
organism like bacteria, fungi, or animal tissue or entire animal it is known as BIOASSAY.

TYPES OF BIOASSAY
QUANTAL:-It is all or none phenomenon
GRADED:-Based on observation that there is a proportionate increase in the observed
response with increase in concentration or dose.

Graded bioassay can be performed by using any of the following techniques

Matching bioassay
Interpolation Method
Bracketing Method
Multiple point bioassay
ANTIMICROBIAL ACTIVITY OF PLANT MATERIALS
Standards of Identity:
Authentication of the crude drug can be done based on the
organoleptic/ morphological and microscopical studies in
comparision with standard texts. but in cases of deterioration due to
storage/ variations with age of the plant other preliminary tests like
extractive values, ash values and chemical tests may be done.
Standards of purity:
Purity may be considered in terms of the presence of authentic
material without any adulteration or admixture. For evaluation of
purity of the drug organoleptic/ morphological and microscopical
studies, histochemical studies, extractive values, ash values and
other physical and chemical methods may be evaluated.
 Standards of potency:
Potency of a drug may be defined as the extent of the activity of
the drug and is checked by using pharmacological models of
living animals and their organs.
It may vary depending upon the type of crude drugs to be checked
for potency. In case of natural bitters like kalmegh, gentian the
bitterness value is compared with bitterness standard quinine
hydrochloride solution, For saponins haemolytic activity is
measured. Astringency of tannins is measured by studying its
ability to bind to standar hide powder.
 Standards of safety:
Although the drug may comply with standards of identity, purity
and potency the drug material may show a toxic reaction.
Toxicity studies for acute, subacute and chronic toxicity studies
must be done to ascertain the safety of the drug.
Standard of microbial limits:
For crude drugs meant for internal use the microbial limits are
Enterobacteriaceae: 103
Salmonella and aerobic bacteria: 105
E.coli: 0
 Senna leaf

Source: Senna consists of dried leaves of cassia senna belonging to the family
leguminosae.
Senna leaf contains not less than 1.0% w/w of sennosides A and B Calculated on the dried
basis.
Identification:
A. Macroscopic: Leaflets- 2.5 to 8cm long and 5-15mm wide at centre, pale yellowish
green, elongated lanceolate, slightly asymmetric at the base, margin entire, apex
acute with a sharp spine. Both the surfaces are smooth with sparce trichomes, odour-
faint and distinctive, taste- mucilagenous and disagreeable but not disctintly bitter.
B. Microscopic: Transverse section shows outer single layered mucilagenous epidermal
cells. Unicellular hairs present. Stomata paracytic, numerous on both surfaces.
Mesophyll consists of upper and lower palisade layers with spongy layer in between,
prismatic crystals of calcium oxalate are present on larger veins.
 C. In the assay, the chromatogram obtained with test solution corresponds to the
chromatogram obtained with reference solution.
 Mobile phase: n- propylalcohol: ethyl acetate: water: glacial acetic acid ( 40:40:29:1)
 Test solution: To 1g of the dried leaves powder add 25ml of methanol and reflux for
10minutes, cool and filter. Reflux the residue with another 20ml of methanol, cool and
filter. Combine the filtrates and evaporate to 10ml.
 Reference solution: To 0.5g of the dried leaves powder add 25ml of methanol and reflux
for 10minutes, cool and filter. Reflux the residue with another 20ml of methanol, cool and
filter. Combine the filtrates and evaporate to 5ml.
 Visualization: UV- 254nm and 365nm /Spray the plate with 20% v/v of nitric acid solution.
Heat the plate at 100-105 for 10 minutes and observe under day light
 Chromatographic profile of test and reference are similar
 Tests
Foreign organic matter: NMT 1%
Total ash: NMT 14%
Acid insoluble ash: NMT2.5%
Loss on drying: NMT 12% - determined on 5g bydrying in oven at 105 ֯ C
Microbial contamination: complies with the microbial contamination tests.
 Assay: Determine by liquid chromatography
 Test solution: To 1g of the dried leaves powder add 10ml of 1% v/v of acetic acid
and 25ml of methanol and reflux for 30minutes, cool and make up the volume to
50ml with methanol and filter.
 Reference solution: A 0.004% w/v solution of sennosides RS in methanol.
 Chromatographic system:
 Stainless steel column packed with octadecylsilane bonded to porous silica
 Mobile phase: A mixture od 82 volumes of 1% v/v acetic acid in water and 18
volumes of acetonitrile
 Flow rate – 1ml/ minute
 Spectrophotometer set at 350nm
 Injection volume-20µl
 Inject the reference solution and the test solution
 Calculate the sennoside content.
 Acacia

 SOURCE: Acacia is the air-hardened, gummy exudation from the

stems of Acacia nilotica, Acacia arabica and other species of
acacia belonging to the family leguminosae.
 Description:
 Tears: Irregular broken pieces of varying size, yellowish white or
yellow or amber in colour with numerous minute fissures, brittle
fractured surface, glossy and odourless
 Powder: A white or yellowish powder , odourless. On treatment
with water it dissolves to give a mucilagenous liquid which is
colourless or yellowish, dense, viscous, adhesive and translucent.
 Identification:
 An aqueous solution is gelatinised by the addition of lead sub
acetate solution.
 To 5ml of 10% solution add gradually, while shaking, 10ml of
ethanol. The cloudy liquid on addition of 0.5ml of acetic acid
gives a white precipitate. Filter and add to the clear filterate
50ml of ammonium oxalate solution, the filtrate becomes
cloudy.
 A 10% solution is either dextro-rotatory or slightly laevo-
rotatory.
 Tests:
 Sterculia gum and agar:
To50 mg of the powdered substance add 0.2ml of freshly prepared ruthenium red
solution and examine under the microscope- the particles do not acquire a ed clour
after irrigation with water.
Agar annd tragacanth: To 10ml of 10% solution add 02ml of leadacetate solution-
no precipitate is produced.
Starch and dextrin: Boil 10ml of a 10% solution and cool, add 0.1ml of 0.05M
iodine- no blue colour is produced.
Tannins: To 10 ml of a 10% solution add0.1ml of ferric chloride solution. A
gelatinous precipitate is formed but neither precipitate nor the liquid shows a dark
blue colour
 Sucrose and Fructose: To 1ml of 10% solution add 4ml of water and resorcinol
and 2ml of HCl and heat on water bath, no yellow or pink colour develops.
 Water insoluble matter: Dissolve 5g in fine powder in 100ml of water in a
250ml, add 10ml of dilute HCl and boil gently for 15 minutes. Filter by suction
while hot through a sintered glass crucible, previously tared, wash throughly
with hot water, dry 105 and weigh. the residue does not exceed 50mg
 Microbial contamination : 1g free from E.coli
 Sulphated ash: NMT 5%
 Acid insoluble ash-NMT 1%
 Loss on drying: NMT 15% determined on 1g by drying in oven at 105֯