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WHO (WORLD HEALTH ORGANIZATION) GUIDELINES FOR STANDARDIZATION

OF HERBAL DRUGS

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 Review Article International Ayurvedic Medical Journal ISSN:2320 5091

WHO (WORLD HEALTH ORGANIZATION) GUIDELINES FOR STANDARDI-

ZATION OF HERBAL DRUGS
Nilakshi Pradhan1, Jyoti Gavali2, Nitin Waghmare3
1
HOD, Shalakyatantra Department, 2Associate Professor, Rasashastra Department,
3
HOD, Kaumarbhritya, Associate Professor;
Shalakyatantra Department, Sumatibhai Shah Ayurved Mahavidyalaya, Pune, Maharshtra, India

ABSTRACT
Herbal drugs are being used as medicines from ancient period. The increased use of herb-
al drugs, and concerns over their safety and efficacy have certainly augmented the need of stand-
ardization of these herbal drugs. WHO has set up guidelines for standardization of these drugs,
which are used as a standard by the majority of countries. The standardization includes the exter-
nal (macroscopy/microscopy) as well as internal examination/ash values, extractive values and
many other parameters to identify, authentify and study its chemical composition. Standardiza-
tion of the medicinal plants will ensure indirectly that the plants are conserved for their medici-
nal and nutritive value. Standardization confirms the safety of the medicinal plant but efficacy
has to be judged clinically or in the laboratory. There is a thin line between efficacy and the
presence of chemical compounds in the drug. The major hurdle in standardization of the batch to
batch variation in the plant compounds. Addition of finer analytical methods of the chemical
compounds may help to minimize the variation and give a better resolution of the plant drug.
Importance of toxicological examination has increased manifolds as contamination can occurs at
various stages, from collection, storage, analysis or processing to extraction of active principles.
These parameters should be recorded for years together; their database should be generated, rec-
orded and analyzed statistically to see the difference in quality and quantity of the chemical
compounds.
Keywords: Standardization, herbal drug, botanical, pharmacological, toxicological, parameters

INTRODUCTION
Medicinal plants are in use for the crude drugs by means of various parameters
purpose of treatment of different ailments like morphological, microscopical, physical,
since centuries. There has been an immense chemical and biological observations is
increase in sales of herbal OTC (Over the called standardization.1Standardisation of
counter drugs. This is growing to a billion the herbal drug begins from the collection of
dollar industry. The need for safety and effi- the herbal drug to its packaging/use as med-
cacy has also escalated since the western icine. The impediments in standardization
interest has grown. Thus the need for stand- of herbal drugs-
ardization has come into view. The process  Variability in the chemical composition
of evaluation of the quality and purity of of the soil and changes in the climate in-
 Nilakshi Pradhan et al: Who (World Health Organization) Guidelines For Standardization Of Herbal Drugs

fluence the range of phyto constituents

individual characteristics in all respects
present in the herbal drugs.2
(Qualitative and Quantitative measure-
 Growing deforestation is leading to in-
ments).Observe identifying characteristics in
crease in the number of endangered spe-
few more slides to confirm the particular
cies of medicinal plants. This leads to
organized crude drug. Compare these char-
addition of adulterants or substitutes to
acteristics with characteristics of the same
the herbal drug. Addition of adulterants
powdered crude drug mentioned in the ref-
and substitutes can change the safety and
erence book.
efficacy of the drug.
PHYSICOCHEMICAL PARAMATERS-
Objectives: The present article will review the
Chromatographic fingerprint5-Separation,
WHO guidelines for standardization of
identification, impurity detection and assay
Herbal Drugs as well as focus on the current
of herbal drug in the formulation or in the
and future trends of the methods used for
extract are carried out by following meth-
analysis of the herbal drug.
ods: -HPTLC, HPLC/Densitometric chroma-
Thus to overcome these obstacles
tography, GLC, TLC
there are many standards and parameters set
Importance-The herbal drug shows variabil- ity
by different pharmacopeias, but guidelines
in its chemical constituents according to
set by the WHO remain most significant.
various locations/weather. To avoid any er-
Materials and Methods
roneous identification chromatographic fin-
BOTANICAL PARAMATERS4-
gerprint remains the assessment of choice.
Sensory evaluation-Visual macroscopy,
Ash values6-The types of ash determined are
Colour, Odour, Taste, Fracture are the
Total ash, Acid insoluble and water sol-
common tests conducted for identification of
uble. Ash value is used to determine the
the crude drug.
quality and purity of the drug and to estab-
Foreign matter-It has to be determined if the
lish its identity. Ash contains inorganic radi-
foreign matter is organic (Moulds, In- sects,
cals lie phosphates, carbonates, and silicates
Animal excreta etc.) or Inorganic (Stone,
of sodium, potassium, magnesium, calcium,
soil etc).Foreign matter is considered as-
etc. These are present in definite amount in a
 material not collected from the original
particular crude drug, hence quantitative de-
plant source (insects, moulds, or other
termination in terms of various ash values
animal contamination)
helps in their standardization. Ash value is
 Parts of the organ, or organs from which
used to determine foreign inorganic matter
the drug is derived other than the parts
present as impurity.
named in the definition and description.
Total Ash Value-The method of total ash is
Methods to determine the foreign organic
designed to determine the amount of materi-
matter-
al that remains after ignition. Ash is classi-
 Manual method
fied as physiological ash which is derived
 Lycopodium spore method
from the plant tissue itself and non-
Microscopy-Identification of histological
physiological ash which is the residue after
characters(under low and high power).Study
ignition of extraneous matter (e.g. sand and
soil).It is carried out at low temperatures

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possibly because alkali chlorides, which are

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 Nilakshi Pradhan et al: Who (World Health Organization) Guidelines For Standardization Of Herbal Drugs

volatile at low temperatures, may be lost.

nary pressure. Many herbal drugs contain
The total ash consists of carbonates, phos-
volatile oil which is used as flavoring agent.
phates, silicates and silica.
For the drugs containing volatile constitu-
Acid insoluble ash- Sometimes, inorganic
ents, toluene distillation method/steam dis-
variables like calcium oxalate, silica, and
tillation method is used to determine the
carbonate content of the crude drug affects
volatile oil contents.
‘Total cash value’. Such variables are re-
PHARMACOLOGICAL PARAMA-
moved by treating with acid (as they are sol- 9
TERS - Bitterness value- Medicinal plants
uble in hydrochloric acid) and acid insoluble
having strong bitter taste are therapeutically
ash value is determined. Acid insoluble Ash,
used as appetizing agents .The bitterness is
Water soluble ash and sulphated ash are also
determined by comparing the threshold bit-
evaluated.
ter concentration of an extract material with
Extractive values7- It is useful for evalua-
that of quinine hydrochloride .The bitterness
tion of a crude drug. It gives an idea about
value is expressed as unit’s equivalent to the
the nature of the chemical constituents pre-
bitterness of a solution containing 1gm of
sent in a crude drug. Useful for estimation of
quinine hydrochloride in 2000ml.
constituents extracted with the solvent used
Method for determination-0.1gm of quinine
for extraction. Employed for material for
hydrochloride is dissolved in 100ml drink-
which as yet no suitable chemical or biolog-
ing water and the stock solution is prepared.
ical assay exists. It can be done by following
Then it is diluted and tested and compared
methods: Cold maceration, hot extraction
with drug.
and ethanol.
Bitterness value in unit per gm = 2000×C÷
Moisture content and volatile matter8-
A×B
The moisture content of the drug should be
Where, A = concentration of stock solution
minimized in order to prevent decomposi-
B = volume of test solution in tube with
tion of crude drug either due to chemical
threshold bitter concentration C = quantity
change or microbial contamination.
of quinine hydrochloride in the tube with
The moisture content is determined by heat-
threshold bitter concentration
ing a drug at 105˚c in an oven to a constant
Haemolytic property- Many medicinal
weight. E.g. – Aloe should have moisture
plant materials, of the families Caryophyl-
content not more than 10% w/w Moisture
laceae, Araliaceae, Sapindaceae, Primula-
content can be determined by following
ceae, and Dioscoreaceae contain saponins.
methods- Gravimetric, Volumetric and in-
The most characteristic property of saponins
strumental.
is their ability to cause haemolysis; when
Gravimetric method-Loss on Drying,
added to a suspension of blood, saponins
Volumetric-Azeotropic Toulene distillation
produce changes in erythrocyte membranes,
method,
causing haemoglobin to diffuse into the sur-
Instrumental- GC, NMR etc.,
rounding medium. The haemolytic activity
Volatile oil content- Volatile oils are the
of plant materials, or a preparation contain-
liquid components of the plant cells, immis-
ing saponins, is determined by comparison
cible with water, volatile at ordinary tem-
with that of a reference material, saponin R,
perature and can be steam distilled at ordi-
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 Nilakshi Pradhan et al: Who (World Health Organization) Guidelines For Standardization Of Herbal Drugs

which has a haemolytic activity of 1000

Foaming index- Many medicinal plant ma-
units per g.
terials contain saponins that can cause per-
Determination- Calculate the haemolytic
sistent foam when an aqueous decoction is
activity of the medicinal plant material using
shaken. The foaming ability of an aqueous
the following formula: 1000 ×a/b
decoction of plant materials and their ex-
Where, 1000 = the defined haemolytic ac-
tracts is measured in terms of a foaming in-
tivity of saponin R in relation to ox blood, a
dex. Calculate the foaming index using the
= quantity of saponin R that produces total
following formula 1000/a : where a = the
haemolysis (g) b = quantity of plant material
volume in ml of the decoction used for pre-
that produces total haemolysis (g)
paring the dilution in the tube where foam-
Astringent property- It is determined by
ing to a height of 1 cm is observed. Saponins
amount of tannins present in the drug
give persistent foam when shaken with wa-
.Tannins (or tanning substances) are sub-
ter. Hence, plant material/extract containing
stances capable of turning animal hides into
saponins is evaluated by measuring the
leather by binding proteins to form water-
foaming ability in terms of foaming index.
insoluble substances that are resistant to pro-
Toxicological Parameters11-
teolytic enzymes. This process, when ap-
Arsenic and heavy metals- Contamination
plied to living tissue, is known as an "astrin-
of medicinal plant materials with arsenic and
gent" action and is the reason for the thera-
heavy metals can be attributed to many
peutic application of tannins. Chemically,
causes including environmental pollution
tannins are complex substances; usually oc-
and traces of pesticides. There are different
cur as mixtures of polyphenols that are diffi-
methods to identify the amount and concen-
cult to separate and crystallize.
tration of heavy metals in herbal drugs.
Determination of Tannins10 -Calculate the
Limit test for arsenic and Limit test for
quantity of tannins as a percentage using the
cadmium and lead are few of them. The con-
following formula: where w = the weight of
tents of lead and cadmium may be deter-
the plant material in grams T1=Weight of
mined by inverse voltametry or by atomic
material extracted in water T 2=Weight of
emission spectrophotometry.
material not bound to hide powder
Determination-The following maximum
T0=Weight of hide powder material soluble
amounts in dried plant materials, which are
in water that bind to standard frieberg Hide
based on the ADI values, are proposed: ▫
powder. [T1-(T2-T0)]×500/w
lead, 10 mg/kg; ▫ cadmium, 0.3 mg/kg. Stain
Swelling Index- The swelling index is the
produced on HgBr2 paper in comparison to
volume in ml taken up by the swelling of 1 g
standard stain.
of plant material under specified conditions.
Pesticide residues12-Examples of pesticide
Its determination is based on the addition of
residues- Chlorinated hydrocarbons and re-
water or a swelling agent as specified in the
lated pesticides: BHC, DDT ,Chlorinated
test procedure for each individual plant ma-
phenoxyalkanoic acid herbicides: 2,4-D;
terial (either whole, cut or pulverized). It
2,4,5-T ,Organophosphorus pesticides:
gives an idea about the mucilage content of
malathion, methyl parathion, parathion,
the drug; hence it is useful in the evaluation
Carbamate insecticides: carbaryl (carbaril),
of crude drugs containing mucilage.
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 Nilakshi Pradhan et al: Who (World Health Organization) Guidelines For Standardization Of Herbal Drugs
Dithiocarbamate fungicides: ferbam, maneb,

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 Nilakshi Pradhan et al: Who (World Health Organization) Guidelines For Standardization Of Herbal Drugs

nabam, thiram, zineb ,Inorganic pesticides:

Determination- IP method: NMT 2 µg/kg of
calcium arsenate, lead arsenate
aflatoxins B1& Total aflatoxins 4 µg/kg
,Miscellaneous: ethylene dibromide, eth-
.USP method: NMT 5ppb of aflatoxins B1&
ylene oxide, methyl bromide, Pesticides of
Total aflatoxins 20ppb.
plant origin: tobacco leaf and nicotine; pyre-
Radioactive contamination-The range of ra-
thrum flower, pyrethrum extract and pyre-
dionuclides that may be released into the
throids; derris root and rotenoids. Includes
environment as the result of a nuclear acci-
total organic chloride and total organic
dent might include long-lived and short-
phosphorous. Determination of pesticides-
lived fission products, actinides, and activa-
Pesticides should not be more than 1%, an
tion products. Microbial growth in herbals is
ARL (in mg of pesticide per kg of plant ma-
usually avoided by irradiation. This process
terial) can be calculated on the basis of the
may sterilize the plant material but the radi-
maximum acceptable daily intake of the pes-
oactivity hazard should be taken into ac-
ticide for humans (ADI), as, recommended
count. The nature and the intensity of radio-
by WHO, and the mean daily intake (MDI)
nuclides released may differ markedly and
of the medicinal plant material.
depend on the source (reactor, reprocessing
ARL=ADI×E×60/MDI×100 where ADI =
plant, fuel fabrication plant, isotope produc-
maximum acceptable daily intake of pesti-
tion unit, etc.). The radioactivity of the plant
cide (mg/kg of body weight); E = extraction
samples should be checked accordingly to
factor, which determines the transition rate
the guidelines of International Atomic Ener-
of the pesticide from the plant material into
gy Agency (IAEA) in Vienna, Australia.
the dosage form; MDI = mean daily intake
DISSCUSSION
of medicinal plant product.
Herbal drug standardization is very im-
Microbial contamination- Contamination either at
portant for the safety and efficacy of the
source or during processing is pos- sible. Maximum
drug. The safety part can be compensated by
possible limits of each or- ganism are given in
performing various analysis on the drug, but
various texts. WHO lim- it for number of micro-
the efficacy should not be judged only by
organisms per gram of material
the presence of the chemical compounds.
There is a very fine line between efficacy
Type of micro- Finished Raw ma-
and presence of compounds. The need of the
organ ism product terial
1 hour is that Clinical/Laboratory analysis
E.coli 10 104
Salmonella - - should be done to establish a relation be-
Total aerobic 103 - tween both of them. Herbal drug standardi-
bacteria zation should be done through multiple
Enterobacteria 103 are naturally
Alfatoxins- Aflatoxins - occur- modes as the concentration of the phyto-
ringmycotoxins produced mainly by Asper- chemicals varies according to climate, soil
gillusflavus and Aspergillusparasiticus. The and environment. The database for the same
presence of aflatoxins can be determined by should be maintained throughout the year
chromatographic methods using standard and the data should be analyzed statistically
aflatoxins B1, B2, G1, G2 mixtures. for proper understanding of the medicinal
plant. Newer aids of research should be used

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 Nilakshi Pradhan et al: Who (World Health Organization) Guidelines For Standardization Of Herbal Drugs
to identify minute variations. Finer analyti-

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 Nilakshi Pradhan et al: Who (World Health Organization) Guidelines For Standardization Of Herbal Drugs

cal methods are now available which can be

4. Kokate C.K., Gokhale, S.B. 2001. Prac-
incorporated to analyze the herbal drugs.
tical Pharmacognosy. 2nd ed. NiraliPra-
Gel electrophoresis of isolated and purified
kashan, Pune, p. 14-19. 2
DNA samples is done to identify the herbal
5. Yvan Vander Heyden, Sept 1
drug as it cannot vary with factors like cli-
2008,Extracting Information fromChro-
mate, etc. DNA fingerprinting can be made
matographic HerbalFingerprints,LCGC
the choice of analysis for a perfect assess-
Europe,Volume 21, Issue 9
ment of authentification. Flow cytometry
6. Quality control of herbal drugs. 4th ed.
can also be applied for herbal drug identifi-
Business Horizons, New Delhi, P. 184-
cation, drug discovery and development.
219.
CONCLUSION
7. Ansari S.H. 2006. Essentials of Pharma-
The WHO guidelines are followed
cognosy. 1st ed. Birla Publications, New
all over the world but the need of the hour is
Delhi, p. 581- 596.
to update these principles with application of
8. http://www.ncbi.nlm.nih.gov/pubmed/18
newer methods of analysis. The herbal drug
396809.
assessment in Ayurveda is about the whole
9. Anonymous, 2010. Indian Pharmacopoe-
drug rather than concentrating on the active
ia. Vol.-3, Government of India, Minis-
principles or phytoconstituents, thus finer
try of Health and Family Welfare, New
methods of standardization should be devel-
Delhi p. 2467-2472
oped. As Ayurvedic drugs are also included
10. Mukherjee P K & Peter J Houghton.
in the Drugs and Cosmetics Act, 1940 the
2009. Evaluation of Herbal Medicinal
drugs have to be safe and effective at the
Products Perspectives on quality, safety
same time. This brings about the need for
and efficacy. Pharmaceutical Press Lon-
finer standardization of herbal drugs. We
don P, 19-23
can certainly avoid the external contamina-
11. Iqbal Ahmad, FarrukhAqil, and Mo-
tion due to the pesticides by organic farm-
hammad Owais. Modern Phytomedicine,
ing, heavy metal contamination by perform-
Turning Medicinal Plants into Drugs
ing soil analysis and other tests, radioactive
WILEY-VCH Verlag GmbH & Co.
contamination is not very common in India
KGaA,Weinheim fGermany 2006 p.26-
but it can be prevented by using healthier
53
ways of prevention.
12. http://testinglaboratoryindia.com/ha-
REFERENCES
analytical-specifications.htm
1. K R Khandelwal, 2013, PracticalPhar-
CORRESPONDING AUTHOR
macognosy, 2nded, Nirali Prakashan,
Dr Nilakshi Pradhan
Pune,23.1-23.4.
Email: sumedhayk@yahoo.com
2. SS Agarwal,M Paridhavi,2007. Herbal
Drug Technology,FirstEd,Universities
Press(India) Pvt. Ltd,Hybd.Pg629-630. Source of support: Nil
3. WHO Quality Control Methods for Me- Conflict of interest: None Declared
dicinal Plant Materials, World Health
Organization, Geneva, 1998

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