Evaluation and

Quality Control of
crude drugs
Purnima V
 Quality can be defined as the status of a drug that is
determined by identity, purity, content and other
physical, chemical and biological properties or
manufacturing processes.
 Quality control refers to processes involved in
maintaining the quality and validity of a manufactured
product.
 In general, all medicines whether they are of synthetic
or of plant origin, should fulfill the basic requirements of
being efficacious and safe and this can be achieved by
suitable clinical trials.
Problems that influence the quality control of crude drugs:
1. Herbal drugs are the mixtures of various constituents'
active principles in most cases are not known. Selective
analytical methods and reference standards are not
available.
2. Substitution and adulteration as a result of
carelessness, ignorance or fraud.
3. Plant materials are chemically and naturally
variable. The process of harvesting, drying, storage
and transportation and processing have an effect.
Phytochemical profile is affected by factors like
season changes, geographical location etc.
 Several herbal pharmacoepias like Chinese
Herbal Pharmacoepia, British Herbal
Pharmacoepia, British Herbal Compendium,
Japanese standards for herbal formulation and
Ayurvedic Pharmacoepia of India.
 These lay down monographs for herbs and herbal
products to maintain their quality in their
respective nations
Standardisation:
Standardization is an important step for the
establishment of a consistent biological
activity, a consistent chemical profile, or
simply a quality assurance program for
production and manufacturing of herbal drugs

Confirmation of identity and

determination of quality, safety and
purity.
Need for standardisation:
 Modern system is based on sound experimental data.
 Pharmacoepial std on crude drugs and finished
products are difficult.
 c-GMP for herbal drugs are not well defined nor barest
minimum stds are maintained or regulated.
 Lack of quality control stds have resulted in mild to
serious adverse effects ranging from hepatotoxicity
and death.
 Herbal ingredients reqiure tools for determining
identity, purity and quality and tools have to be
sufficient, rapid and cost effective with GMP
reqiurements.

WHO has set specific guidelines for assesment of safety,

purity, efficacy and quality of herbal medicines.
The quality control of a traditional medicine
1. The traditional methods are procured and
studied, and documents and the traditional
information about the identity and quality
assessment are interpreted in terms of modern
assessment or monograph in herbal
pharmacopoeia
2. The crude drug can be evaluated or identified
by five methods:
 Evaluation of Crude Drugs
Means to identify and to determine quality,
safety and purity.
1. It has to be certain of identity of the collected
plant from proper source by matching to
authentic plant sample (Authentication)
2. Authentication is done at Agarkar Institute, Pune
3. The samples are stored in herbarium with
unique voucher no given to the preserved
sample for future reference
Evaluation of drugs involves the
following methods:
1. Organoleptic
2. Macroscopy & Microscopic
(Botanical)
3. Chemical
4. Physical
5. Biological
I- ORGANOLEPTIC EVALUATION
Organoleptic refers to evaluation by means of the
organs of sense which includes:
- The macroscopic appearance of the drug,
- Its odour and taste and the feel of the drug to
the touch.
Description of the macroscopic characteristics of a
drug include:
1. Shape and size.
2. Colour and external markings
3. Fracture and internal colour.
4. Odour and taste.
Examples:
1. Shape of drug may be cylindrical (sarsapilla),
subcylindrical (podophyllum), conical (aconite), fusiform
(jalap) etc
Size represent length, breadth, thickness, diameter etc.
2. Color means external color which varies from white to
brownish black are important diagnostic characters.
3. The general appearance (external marking) of a crude
drug often indicates whether it is likely to comply with
prescribed standard like furrows (alternate depression or
valleys), wrinkles (fine delicate furrows), annulations
(transverse rings), fissures (splits), nodules (rounded
outgrowth), scars (spot left after fall of leaves, stems or
roots).
1. Cylindrical Shape of
Sarpasilla

2. Subcylindrical Shape of
Podophyllum

3. Conical shape of Aconite

4. Fusiform shape of Jalap

Shapes of Leaves
4.Aromatic odour of umbelliferous fruits and sweet taste
of liquorice are the examples of this type of evaluation
where odor of drug depends upon the type and quality of
odourous principles (volatile oils) present.
Taste is specific type of sensation felt by epithelial layer
of tongue. It may be acidic (sour), saline (salt like),
saccharic (sweetish), bitter or tasteless (possessing no
taste).
5. The fractured surfaces in cinchona, quillia and cascara
barks and quassia wood are important characteristics
Splintery, Granular, vitreous, Fibrous
II-MICROSCOPIC EVALUATION
 The microscope is essential in the identification of
powdered drug and in the detection of the
adulterants in powdered plant OR animal drugs.
 Microscopical description of the drug in sectional
view and powdered form is listed in official
monograph .
 Every tissue posseses characterstic tissue
structure, demonstrated through study of tissue
arrangement, cell wall and configuration when
properly mounted in stains, reagents and
media.
 Lignin: Red with phlorglucinol and Hcl
 Mucilage: Pink with Ruthenium red
 Starch and Hemicellulose: N/50 Iodine
 Sudan red-III for oil detection, which stains red
 Ferric chloride for tannins- black/blue/brown
 Chloral hydrate solution is useful to remove
chlorophyll
1. Histological studies on very thin transverse (TS) or
longitudinal (LS) sections properly mounted in suitable
stains, reagents or mounting media.

2. The characteristic feature of cell walls, cell contents,

starch grains, calcium oxalate crystals, trichomes, fibres,
vessels etc are studied.
Eg:
1. Presence of pith in rhizomes and absence in roots
2. Warty trichomes of Senna. Calcium Oxalate crystal
Sheet.
3. Clove stalk contains sclerides and Ca oxalate and
clove powder does not contain the two.
Plant tissue types:
a. Ground tissue comprises three important cells, i.e.,
parenchyma, collenchyma, and sclerenchyma cells. Three
ground tissue cells are responsible for storing food and water,
supporting the plant as its support system, and in-plant
photosynthesis.
b. Vascular tissue - Cells like phloem, xylem, and cambium
combinable made these tissues that help transport minerals, food,
and hormones in the plant cell.
Xylem -Required for transportation of water
Phloem -Required for transportation of food
c. Dermal tissue - These tissues are made up of epidermal cells
that lie on the surface of the plants and prevent loosing of water
from the plant.
Epidermal cells with stomata
and trichomes
Transverse Section of Bark
Cinchona Bark
Histology of bark
POWDER CHARACTERSTICS
T.S of leaf
Powder characterstics
Types of trichomes
1 Covering trichomes
a. Unicellular tichomes Nuxvomica, Senna

b. Multicellular-unbranched trichomes
(i) Uniseriate Datura
(ii) Biseriate Calendula officinalis
(iii)Multiseriate Male fern

c. Multicellular-branched trichomes Verbascum

thapsus
2.Glandular trichome
a. Unicellular glandular trichome Vasaka
b. Multicellular glandular trichome Digitalis
purpurea
3.Hydathode trichome - Piper betal
There are several types of stomata, distinguished by the
forms and arrangement of the surrounding cells.
 Anomocytic (irregular – celled) /Ranunculaceous
 Anisocytic (unequal – celled) /Cruciferous
 Diacytic (cross- celled) /Caryophyllaceous
 Paracytic (parallel celled) /Rubiaceous
Calcium oxalate crystals
Several inorganic components like calcium oxalate crystals,
calcium carbonate crystals and silica are frequently found
in plants.
They are crystalline in nature.
Specific shape of calcium oxalate crystals can be used for
the identification of drugs.
Due to this reason they are known as diagnostic characters
of the plant.
Function : protein metabolism gives oxalic acid, which is
harmful to plants. To remove harmful effect of oxalic acid,
plant forms harmless calcium oxalate crystals with calcium
ion. (obtained from soil)
 Deposited in different tissues, in different forms.
Harmless to plant. Doesn’t take part in metabolism,
hence called excretory product.
Significance:
1)They give protection to the plant against birds and
animals.
2)They have great diagnostic value.
3)Presence and absence of crystals, type of crystals and
dimensions are useful in correct identification of crude
drugs.
4)Help in detection of adulterants.
Clove stalk contains calcium oxalate prisms but clove
flower bud does not
2. Microscopic linear measurement : Leaf
constant determination and quantitative
microscopy are also used in this evaluation

 Linear measurement include size of starch

grains, length and width of fibres,
trichomes etc using micrometry
 Leaf constants include stomatal no,
stomatal index, vein islet, vein
termination, Palisade Ratio
 Camera lucida (Latin: “light chamber”)
 Optical instrument patented in 1806 by William Hyde
Wollaston to facilitate accurate sketching of objects.
 Camera lucida is an optical device which when connected to a
microscope can help a person to draw the image of an object
in scale with the actual object by tracing on the superimpose
image.

 The principle is very simple, by looking into the prisms placed

at right angle, two images will enter the eye; one of the object
to be traced and other of the paper.
 The resulting effect is that your eye perceives illusion of seeing
objects in front of the instrument on the drawing surface
beneath.
Swift Ives Abbes
 Leaf Constant
 Stomatal Number: Average number of stomata per sqmm of the epidermis of
the leaf
 Stomatal Index: Percentage which the number of stomata form to total
number of epidermal cells, each stoma being counted as one cell.
SI= S/ S+E X100

Stomatal number varies considerably with the age of the leaf and
due to the changes in environmental conditions, stomatal index is
relatively constant and therefore, of diagnostic significance for a
given species.
Stomatal index is employed for the differentiation of allied or
closely related species of the same genus in air dried, as well as,
fresh conditions
STOMATAL NO AND INDEX

Assignment: Stomatal no and index of

Senna leaves on both the surfaces
 Vein islet number: Number of vein islets per sqmm of
the leaf surface midway between the midrib and the
margin
 Vein Termination number: It is defined as number of
vein terminations per sqmm of the leaf surface midway
between the midrib and the margin
 Palisade Ratio: Avg number of palisade cells beneath
each epidermal cells.
Unlike vein islet number for the determination of
which unbroken surface is required, palisade can be
performed on the powdered drugs.
VEINISLET AND VEIN TERMINATION

PALISADE RATIO
Quantitative Microscopy
Lycopodium Spore Method
 Determination of proportion of substances present is done
by the means of microscope, using lycopodium spore
method
 Starch or starchy drugs when used as adulterants , can be
determined by number of starch grains per mg and
calculating the amount from the known number of starch
grains per mg of pure starch
 Standard ginger contains 2,86,000 starch grains/mg

 The number of diagnostic character of each ingredient

is directly proportional to quantity of the drug. (Can be
done for Stone cells etc)
 Support to the chemical method of standardization.
 Lycopodium is used in quantitative microscopy as
a reference standard because of its
characteristic shape, appearance & uniform
size (abut 25micrometer)
 It is neutral and does not encapsulate with any
other character of a powder.
 Lycopodium powder contains on an average
94,000 spores per milligram.
 A powdered drug can be evaluated by this
method if it contain :
 Well defined particles which may be counted
eg: Pollen grains or starch grains.

% Purity = N x W x 94,000 x 100

SxMxP
 Percentage purity of ginger can be
calculated using following equation
 % Purity = N x W x 94000 x 100
SxMxP
Where,
N: Number of starch grains in 25 fields
W: Weight in mg of lycopodium taken
S: Number of spores in the same 25 fields
M: Weight in mg of the sample calculated on the basis if dried sample at
1050C
P: 2, 86,000
94,000: Number of lycopodium spores per mg
Pure Jamaica ginger contains two lakh 86 thousand starch grains per mg
Chemical Evaluation
 Most of drugs have definite chemical constituents to
which their biological or pharmacological activity is
attributed.
 Preliminary phytochemical screening is a part of
chemical evaluation.
 These qualitative chemical tests are useful in
identification of chemical constituents and detection of
adulteration.
 Chemical evaluation includes qualitative chemical
test, quantitative chemical test and instrumental
analysis
 Qualitative tests include identification test for
various phytoconstituents like alkaloids,
glycosides, tannins etc.
Alkaloids: Dragendroffs reagent gives orange ppt
Anthraquinones: Borntangers test gives pink colour
Tannins: Fecl3 gives green colour.
Gold beater Skin test:
Test to distinguish between True Tannins and Pseudo
Tannins
Goldbeaters skin: Prototype of untanned fresh skin of
animal & is obtained from membrane of intestine of ox.
Membrane + Hcl , Rinse with d/w, Tannin solution (5
mins), washings with d/w+ Ferrous sulphate
Brown or Black Colour on skin (Tanning)
+ ve True Tannins confirmed
-ve /partial Pseudo Tannins
Quantitative chemical Test:
Acid value, Saponification value: Lipids, Balsams,
resins etc.
Acetyl Value: Volatile oils
Ester value: Balsams
Determination of volatile oil : Distillation method.
Clavengers apparatus
 Chemical Assay include assay for alkaloid, resin,
glycoside, vitamins and other constituents
 The results obtained can conclude presence of inferior
or exhausted drug .
 Quantitative estimation of individual phytochemicals by
using chromatography, spectroscopy, etc. are
parameters of quantitative chemical evaluation
Instrumental Analysis:
Chromatographic methods include: Paper chromatography,
LC, HPLC, HPTLC etc

Spectroscopic methods include: UV and Visible, IR, NMR.

Physical Evaluation
Physical methods include: Solubility,
Specific gravity, optical rotation, viscocity,
refractive index, melting point, water
content etc.
Solubilty:
Colophony: light pet ether
Castor oil: Alcohol
Alkaloid bases: org solvent
Alkaloidal salt: Aqueous.
Optical rotation: Optically active substances
Dextrorotatory (enantiomer which rotates plane polarised
light to right)
Levorotatory (enantiomer which rotates plane polarised light
to left)
Eg: Eucalyptus oil(0 to +10°)
Honey (+3° to -15°)
Refractive Index:
The ratio of the velocity of light in air to its velocity in the
substance (drug) is termed as refractive index of the
substance
Refractive index is constant for liquid depending upon purity
and it is considered for its standardization
Eg: Castor oil: 1.4758, clove oil: 1.527 to 1.535 etc.
Specific gravity: relative density.
The ratio of the mass of a solid or liquid to the mass of an
equal volume of distilled water under prescribed conditions
of temperature and pressure.
Cottonseed oil: 0.88 – 0.93
Coconut oil: 0.925
Castor oil: 0.95 etc.
Viscosity: resistance of a fluid to flow.
Viscosity of a liquid is constant at given temperature and is
the index of its composition.
Melting Point: M.P of a solid is the temp at which it changes
state from solid to liquid.
In case of pure chemicals or phytochemicals, melting points
are very sharp and constant.
The drugs from animals and plants origin contain the mixed
chemicals; they give certain range of melting point.
The purity of crude drugs can be ascertained by determining
their melting points in the range.
Bees wax: 62 - 65°C
Agar: 85°C
Boiling point:
This parameter is applicable to all liquid phytochemicals
like oils or few alkaloids.
Shift in boiling point range helps in determining purity of
phytochemicals

Moisture Content:
Moisture content of drug is responsible for decomposition of
crude drugs by producing chemical change or microbial growth.
Loss on Drying: Moisture content is determined by heating a drug
at 105°C in an oven to constant weight
Eg: Digitalis 5%w/v, ergot: 8%w/v
Other: Volumetric Azeotropic distillation method, Karl Fischer
method.
ASH VALUE
 The ash values mainly represent the inorganic
residues such as phosphates, carbonates and
silicates present in herbal drugs.
 These are one of the major indices to illustrate the
quality as well as purity of herbal medicine.
 Remnants of crude drugs after incineration contains
mostly inorganic salts known as Ash. It varies in
case of many crude drugs. Its study gives an idea
about the quality and purity of the drug during
evaluation.
Ash Values:
Determination of ash is useful for determining low grade
products and excess of sandy or earthy matter.
Different ash valus: Total ash, Acid insoluble ash, water
soluble ash, sulphated ash.
Total Ash: 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.
The maximum temp for carbon removal for total ash
detection should NMT 450° because alkali chlorides may
be volatile in higher temp would be lost.
Total Ash contains mainly oxides, sulphates,
phosphates, silicates and chlorides
Acid insoluble ash: Ash insoluble in dil Hcl.
Majority of drugs contain calcium oxalate and quantity of
Ca oxalate varies. Total ash of crude drug vary within wide
limits for specimens of genuine drug.
Eg: Rhubarb, total ash range from 8 to 40%, such cases
total ash is useless to detect earthy matter. Acid insoluble
ash is preferable.
Ca oxide or carbonate yielded by the inceneration will be
soluble in Hcl
By this detection of presence of excessive earthy material
which is likely to occur in roots and rhizomes.
Water Soluble Ash:
Water soluble ash is the difference between total
ash and water insoluble residue.
Detect the presence of material exhausted by water.
The total ash and water soluble ash values of ginger are 6
and 1.7%resp
The ash that dissolves in water, providing insights
into the soluble mineral content of the plant

Sulphated ash is done to produce consistent ash.

Produced by addition of sulphuric acid before ignition
and then incinerated in order to get sulphated ash. The
temp used for this is above 600°C.
All oxides and carbonates are converted into sulphates.
Extractive Values:
The crude drugs have their biological activity due to
chemical constituents. These constituents are soluble in
different solvents.

It is the amount of active constituents extracted with

solvents from a given amount of medicinal
plant material.

Extracts are obtained by exhausting the crude drugs with

different solvents .
Water soluble extractive is used for drugs containing
water soluble constituents like glycosides, tannins,
mucilage etc

Ether Soluble extractives for drugs containing volatile

oils and fats

Alcohol soluble extractive: Alcohol is an ideal solvent for

extraction of various chemicals like tannins, resins, etc.
95% ethyl alcohol is used for determination of alcohol
soluble extractive
Foreign Organic Matters:
Parts of the organ or organs other than those parts of drugs
in the definition and description of drug are known as
foreign organic matters.
Insects, moulds, earthy matter, animal excreta etc.
Each and every vegetative drug has their own limits.
Garlic should not contain more than 2%
Shatavari NMT 1%
Rf value
 The active constituent present in the crude drugs when
subjected to thin layer chromatographic study, move on
the TLC plates according to their affinity with the
solvent.
 The quality control employs TLC for assessing the
quality and purity of the drug.
 Rf value is the ratio of distance moved by the solute
divided by the distance moved by the solvent front.
 It varies from zero to one
Biological Evaluation
Some drugs have specific biological and pharmacological
activity which is utilized for their evaluation.
Actually this activity is due to specific type of
constituents present in the plant extract.

For evaluation the experiments were carried out on both

intact and isolated organs of living animals.
With the help of bioassays (testing the drugs on living
animals), strength of drug in its preparation can also be
evaluated.
Toxicological determination: Presence of heavy metals,
Aflatoxins, Microbial count etc.
Microbial assays are also part of biological evaluation
Swelling Index:
Defined as volume in ml occupied by 1g of drug after it has
swollen for 4 hours.
Procedure: Drug + 25ml water in graduated cylinder, shaken
every 10 min for 1 hr and then allowed to stand.
Whole drug or powdered drug can be used.
Swelling factor: standing for 24 hrs.
Swelling power: in respect to Isaphgul husk.
Isaphgul husk : NLT 40 (0.1g)
Isaphgul seed: NLT 9
Bitterness Value: Bitterness value is determined
organoleptically by comparison with a quinine
hydrochloride solution which acts as a standard.
Foaming Index:
 Weigh 1 g of coarse powder of the drug and transfer to
500ml conical flask containing 100ml of boiling water.
Maintain at moderate boiling for 30 minutes. Cool and
filter into 100ml volumetric flask and add sufficient
water to 100ml with d/w.
 Pour the decoction into 10 stoppered test tubes in
successive portions of 1ml, 2ml, 3ml etc upto 10ml.
Adjust the volume of liquid in each test tube with
water to 10ml. Stopper the tubes and 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.
Biological testing of herbal drugs:
In standardization/evaluation of herbal drugs, assessment
of biological efficacy is found to be most assuming
method.
(a) Hepatoprotective activity:
Male/female albino rats are used.
Inducing chemical : Allyl alcohol, cause liver necrosis,
Paracetamol induced liver damage, Carbon Tetrachloride
Standard drug : Silymarin
2. Anti-diabetic activity :
Animal – Mice or Rat
Inducing chemical – Alloxan, Streptozotocin Standard drug
– Glipizide, Glibenclamide

3. Anti-ulcer activity :
Animal – Rats
Inducing chemical – Indomethacin, Immobilization stress
Standard drug – Omeprazole
4. Anti-inflammatory activity :
The drugs from plant origin cause anti-inflammatory
effects and used in conditions like rheumatoid arthritis,
gout, etc.
The principle underlying the testing of anti-inflammatory
activity is the reduction of local oedema induced in rat or
Mice paw by injecting irritant, inflammatory substances
like Formaldehyde, Carrageenan, Egg Albumin, Dextran,
Ultraviolet Light

Standard drug – Diclofenac, Indomethacin,

Microbiological Assay
Drug substances that either supress or influence the
growth of microorganisms are analysed by microbial
method.
 Cylinder Plate method
 Turbidimetric Method
Isolated Organ method: Guinea Pig ileum : To test ANS
drugs.
WHO Guidelines for Q.C of herbal drugs
A typical monograph for herbal drugs as per WHO guidelines is
as follows:
Monograph Title:
1. Botanical
Sensory Evaluation:
Visual Macroscopy/Touch/Odour/Taste
Foreign Matter: Foreign plants, animals, minerals etc.
Microscopy: Histological observation, Histochemical
detection, measurements etc
2. Physicochemical
TLC
Ash: total, acid insoluble, water soluble
Extractable matter: Alcohol, water
Water Content & Vol matter: LOD, Azeotropic
Vol oil: Steam distillation

3. Pharmacological
Bitterness Value: Units eq to bitterness of std solution of
quinine Hcl
Haemolytic Activity: On Ox blood by comparing with std ref
saponin
Astringency: Fraction (Tannins) that binds to std hide powder.
Swelling Index: In water, Isaphgul ref
Foaming Index: Foam height produced by 1 gm of material
under specified conditions
4. Toxicological :
Pesticide residue: total organic chloride and total
organic phosphorous
Arsenic: Stain produced on HgBr2 paper in comparison
with std stain
Heavy Metals: Cadmium and lead
5. Microbial Contamination:
Total viable aerobic count
Pathogen: Enterobacteriaceae, E coli
Salmonella, P aerogenosa, S aureus
Aflatoxins: by TLC using std Aflatoxins mixture.
6. Radioactive Contamination