Download Report

Khatoon | Quality control tools for Unani drugs
INTERNATIONAL JOURNAL OF ADVANCES IN
PHARMACY MEDICINE AND BIOALLIED SCIENCES
An International, Multi-Disciplinary, Peer-Reviewed, Open Access, Indexed, Triannually Published Journal
|www.biomedjournal.com|
Macro-microscopy & planar chromatography-Important tools for quality control
and identification of adulterants/substitutes of Unani Drugs
Sayyada Khatoon.
Pharmacognosy & Ethnopharmacology Division, CSIR-National Botanical Research Institute, Lucknow-226001, India.
REVIEW ARTICLE
ABSTRACT
ARTICLE INFORMATION
Article history
Received: 1 April 2015
Revised: 15 April 2015
Accepted: 18 April 2015
Early view: 20 April 2015
*Author for correspondence
E-mail: [email protected]
[email protected]
Q
R
C
o
d
e
Unani drugs of plant origin are herbaceous whole plant or their parts. The ever
increasing demands of these drugs are leading to the adulteration and
substitution of genuine drugs and the poor quality of Unani products. In India,
the crude drug supplies are usually obtained through various trade channels
and are generally lacking in uniform quality. It is very difficult to authenticate the
commercial crude drugs because these are available as dried whole plant or
some part of it. The Macro-microscopy & Planar chromatography (TLC/HPTLC)
are the basic and important tools for proper identification of
adulterants/substitutes of Unani drugs and their quality control.
The macroscopy includes organoleptic characters while the microscopy
encompasses the detection of the cell type and cell contents as well as the
arrangement of cells in tissues. Banafshan, Bhuamla, Pershiaoashan, Rehan,
Resha Khatmi and Zarnab can easily be differentiated from their
adulterants/substitutes by observing the surface characters or comparing leaf
surface microscopy. Similarly, the arrangement pattern and size of fibres, stone
cells, crystals, secretary canals etc in phelloderm and phloem region are valuable
parameters for the identification of most of the bark drugs.
However, the quality assurance of Unani drugs still remains a challenge
because of the high variability of chemical components viz. alkaloids, phenolics,
terpenoids, steroids, glycosides etc. The variation may be diurnal, dioecy,
chemotypic, genotypic, ecotypic, seasonal etc. Fingerprint analysis using
TLC/HPTLC has become the most potent tools for quality control of Unani
medicines because of its simplicity, reliability, rapidity and economy. Not only
the general finger print profile but also chemical reference markers can be
identified and estimated for quality evaluation and authentication of
adulterants/substitutes of Unani medicine viz. asarone in Acorus, phyllanthin &
hypophyllanthin in Phyllanthus species, berberine & tinosporaside in Tinospora,
glycyrrhizine in Glycyrrhiza, gallic and ellagic acids in Terminalia species etc.
Biomedjournal © Copyright 2013, All rights reserved. Biomedjournal Privacy Policy.
INTRODUCTION
In traditional systems of medicine, drugs of plant, animal
and mineral origin are used in their natural or so called
„Crude‟ forms singly or in their mixture or in
combination, to make a formulation (Khatoon et al.,
2006a). Nearly 90 % of the Crude Drugs are obtained from
the plant sources which may be whole plant especially of
herbaceous plants; otherwise their parts such as Root,
Rhizome, Stem, Wood, Bark, Leaf, Flower, Anther,
Pollen, Seed, Fruit and their Exudates or Gums etc
constitute single drugs in all Traditional Systems of
Int J Adv Pharmacy Med Bioallied Sci. 3, 1, 2015.
Medicine even in Unani. The ever increasing demand of
herbal drugs led to a spurt of large-scale commercial
production with multicrores Rs/$ investments in many
countries including India (Aneesh et al., 2009). With the
ever-increasing demand of medicinal plants the supply
line is adversely affected causing adulteration and
substitution for genuine drugs. Such adulteration and
substitution lead to the poor quality of herbal products
(Khatoon et al., 2006a; Mehrotra et al., 2001). It is very
difficult to authenticate the commercial crude drugs as
these are available in the form of dried whole plant or
www.biomedjournal.com
58
Khatoon | Quality control tools for Unani drugs
some part of it. WHO has emphasized on the need to
ensure the quality of medicinal plants/products
(Anonymous, 2003). Countries like China have taken
various measures to check the adulteration by raw drug
standardization, domestication and cultivation of
medicinal plants to ensure sustained supply of quality
plant material to production centers (Li et al., 2011;
Zhong et al., 2014). In India, the supplies are usually
obtained through various trade channels and are
generally lacking in uniform quality. Further, due to the
enormous demand of herbal drugs resulted in depletion
of natural resources is also one of the main causes for
substitutions/adulterations (Khatoon et al., 2006a;
Mehrotra et al., 2001).
Adulteration may broadly be defined as admixture or
substitution of genuine article with spurious inferior,
defective or otherwise useless or harmful substances.
Adulterants
are
those
herbs/materials
having
resemblance with that of genuine herbs/materials but
have no biological or very non significant medicinal
activity while substitutes are those herb/material having
more or less similar medicinal properties of that of
genuine one viz. Shanjaar/Ratanjot - Arnebia nobilis is
genuine drug and A. euchroma, Onosma hispidum can be
used as substitutes but Jatropha curcus is an adulterant
(Khatoon, 1991).
Adulteration/substitution may be deliberate or undeliberate. The Unani and Ayurvedic nomenclature of
drug, traditional and regional names, various vernacular
names of single plant species, are important factors of
un-deliberate adulteration (Khatoon et al., 2006a).
There are several controversial drugs where botanical
identity of plant sources has not been established (Sagar,
2014; Kunle et al., 2012, Mehrotra et al., 2001).
Sometimes not only the various species of a particular
genus but entirely different plant taxa are being sold or
used under the local names (Nizami and Jafri, 2006,
Mehrotra et al., 2001). Another un-deliberate aspect of
lack of quality of plant based drugs is „Natural variation‟
in therapeutically active constituents which are either
primary or secondary metabolites (Pavarini et al., 2012;
Kroymann, 2011; Kale, 2010). The expression of many of
these compounds particularly those of the secondary
metabolites are controlled and conditioned by a variety
of factors such as habitat of the plant, agro climatic
conditions, diurnal, seasonal and also the age and stage
of plant/ part, dioecy, genetic predisposition, etc. Some
of the examples are:
 Asgand (Withania somnifera)- Two varieties of
Asgand have been mentioned in classical Unani
literatue i. e. Asgand Nagori and Asgand Dakani.
Asgand Nagori is preferred for its more potential
medicinal properties (Anonymous, 1982, Ghani,
1920). However, researches showed various
chemotypes with phenotypic variations and
distinct pharmacological activities (Tuli and
Sangwan, 2009; Kumar et al., 2007).
 Rewand-chini (Rheum emodi) should be collected
in
summer
because
anthraquinones
(therapeutically active constituents) were
Int J Adv Pharmacy Med Bioallied Sci. 3, 1, 2015.
present in this season but in winter anthranols
(inactive) were present (Anonymous 1999).
 Degitelis (Digitalis purpurea) showed diurnal
variation in therapeutic active constituent in
leaves which contain more active glycoside at
day time.These glycosides breakup into aglycone
and sugar, a less active substance at night
(Rowson, 1961).
 Dirmanah
(Artemisia
maritima)
contains
maximum therapeutic active constituent santonin in unexpanded flower buds but it is
absent in open flowers (Nadkarni, 1996).
The deliberate adulteration of therapeutically important
plants with a cheaper material is very common to gain
more profit. Various species of a particular genus or
similar looking entirely different plant taxa are being sold
under the same vernacular name or partially mixed with
the genuine drugs. Substances like cloves, fennel,
caraway, which are used to obtain volatile oils by steam
distillation are mixed with genuine articles after removal
of their volatile oils contents. In case of plant based
drugs it is not limited to substitution of one plant to
another or exhausted products, but various traditional
medicines have been found to contain undeclared
synthetic materials or even clay (Khatoon et al., 2006a;
Mehrotra et al., 2001).
The identification of crude herbal drugs differ from that
of living plants since the herbal drugs are usually in a
dried form and may also have been subjected to other
processing procedure. It is also a well known fact that
the therapeutic activity of a medicinal plant is due to the
presence of certain biologically active chemical
constituents. The old literature of Traditional Indian
System of Medicines like Unani and Ayurveda etc.
provided specific instructions for collection by indicating
location/edaphic conditions, habitat, seasonal and even
the stage of the plant/part, growth and developmental
stage of herb (Ibn-Al-Bitar, 1985; Sharma, 2001; Tewari
et al., 2014).
The Macro-microscopy & Planar chromatography
(TLC/HPTLC) are the basic and important tools for proper
identification of adulterants/substitutes of Unani drugs
and their quality control. Some of the important drugs
are discussed in this paper under the heads macroscopy,
microscopy and planar chromatography.
1. Macroscopy
The macroscopic study includes organoleptic characters
i.e. the occurrence, size, shape, colour, surface
markings, margins (leaf), texture, fracture, internal
appearance, cut surfaces, odour and taste of the crude
drug. These external characters also play an important
role for the identification of adulterants/substitutes of
herbal drugs (Mehrotra, 2003; Khatoon et al., 2006a;
Prakash et al., 2013).
i. ‘Banafsha’
Flowers of Viola odorata L. are the official drug of
„Banafsha‟ but other species of Viola viz. V. pilosa and V.
betonicifolia are also commonly being sold as „Banafsha‟.
All three species of Viola can easily be identified by their
macroscopical characters especially by observing the
www.biomedjournal.com
59
Khatoon | Quality control tools for Unani drugs
style and stigma. Style is inflated, broad and smooth in V.
odorata; subclavate, subtruncate and shortly beaked at
the apex in V. pilosa; geniculate at base and clavate
above in V. betonicifolia. However, stigma is decurved &
hairy in V. odorata; truncate with sub-lateral opening in
V. pilosa while convex, forming hump like structure with
lateral side opening in V. betonicifolia (Mehrotra et al.,
1998).
ii. ‘Bhuamla’
Previously Phyllanthus niruri was reported as „Bhuamla‟
in India but after 1985 it was distributed in three species
P. amarus, P. fraternus and P. debilis (Mitra and Jain,
1985). During market surveillance of herbal drugs, it was
observed that almost all the commercial samples, either
comprise of Phyllanthus amarus Schum & Thonn. or P.
maderaspatensis Linn. or mixture of P. amarus, P.
fraternus Webster. and P. maderaspatensis (Khatoon et
al., 2006b). These species can be identified on the basis
of following morphological characters:
 P. amarus- Branchlet 2-6 cm long; leaves 10-20,
elliptic, oblong to ovate, obtuse or minutely
apiculate at apex; flowers axillary, proximal 2-3
axils with unisexual 1-3 male flowers and all
succeeding axils with bisexual cymules, sepals 5
(Fig. 1A).
 P. fraternus- Branchlet 2-11 cm long; leaves 1030, elliptic oblong rounded at the apex; flowers
in axillary, unisexual cymules, proximal 3-4 male
flowers, succeeding solitary female flowers,
sepals 6 (Fig. 1B).
 P. maderaspatensis - Branchlet absent; leaves,
rounded truncate or somewhat obcordate at the
apex, mucronate, much tapering into a very
short petiole; flowers axillary, male flowers
minute in small clusters, sub-sessile; female
flowers solitary and larger with short stalk,
sepals 6 (Fig. 1C).
with medium-hard fracture (Khatoon et al.,
2008a).
2. Microscopy
The microscopical character encompasses the detection
of the type of cells and cell contents as well as the
arrangement of cells in tissues.
i.
Bark drugs
During the drying process the bark drugs develop some
sort of markings, cracks on the surfaces and sometimes
occupy a different shape e.g. flat, curved, recurved,
quilled etc. The arrangement pattern and size of fibres,
stone cells and crystals in phelloderm and phloem region
are valuable parameters for the identification of most of
the bark drugs (Khatoon and Mehrotra, 2009). For
example- „Asok‟- Saraca asoca L. is official drug but
Indian herbal drug markets are full of several
adulterants/substitutes. These can easily be identified
under the microscope- TS S. asoca shows phloem with
small scattered groups of fibres and stone cells, unibiseriate medullary rays while in S. declinata phloem
with solitary or small groups of fibres, presence of fibre
sclereids, uni-triseriate medullary rays; in Polyalthia
longifolia phloem with broad concentric bands of fibre
patches, interrupted by broad radiating multiseriate
medullary rays embedded with mucilage canals; in
Bauhinia variegata with scattered solitary fibres and
calcium oxalate crystals and Shorea robusta with large
scattered stone cells and interlocking medullary rays
(Figure 2) (Khatoon and Mehrotra, 2009)
Saraca asoca
S. declinata
Bauhinia variegata
Polyalthia longifolia
Shorea robusta
Figure 2. TS of stem bark of Asok.
A. P. amarus
B. P. fraternus C. P. maderaspatensis
Figure1. Macroscopy of Phyllanthus species
iii. ‘Resha Khatmi’- Drug consists of the roots of
Althaea officinalis L. (Fam. Malvaceae). Due
to the limited distribution of this species the
roots of another species Alcea rosea L. syn.
Althaea rosea is also being sold in Indian
market. Both the species can be identified
by external characters. The roots of Althaea
officinalis
are
strongly
longitudinally
furrowed, often spirally twisted with shortmedium fracture but in Alcea rosea these
are finely longitudinally furrowed, straight
Int J Adv Pharmacy Med Bioallied Sci. 3, 1, 2015.
60
ii.
‘Pershiaoashan’
It is a drug from Pteridophytes or fern group of plant
kingdom. Adiantum capillus-veneris L. is the official drug
but other abundantly distributed and similar looking
Adiantum species viz. A. lunulatum Burm., A.
peruvianum Klotzsch., and A. venustum D. Don are
commonly found as substitute/adulterant. The salient
distinctive characters under the microscope are the
presence of slightly wavy elongated epidermal cells in A.
capillus-veneris; epidermal cells strongly wavy in A.
lunulatum; star shaped epidermal cells in A. peruvianum;
stomata on lower surface of pinnule but on both the
surfaces only in A. venustum. In addition, rachis anatomy
showed different cellular and stellar characteristics as
identifying characters of aforesaid four Adiantum species
(Singh et al., 2013).
www.biomedjournal.com
Khatoon | Quality control tools for Unani drugs
iii.
‘Rehan’
Ocimum sanctum L. of family – Lamiaceae is the official
drug „Rehan‟ but dried leaves of other Ocimum species
viz. O. basilicum L. , O. canum Sims, O. gratissimum L.
are also being used or sold as drug „Rehan‟. All the
aforesaid Ocimum species can easily be differentiated by
visualizing the surface after removal of chlorophyll
(boiled the leaves with saturated chloral hydrate
solution, stained with safranin and mounted in glycerol).
Ocimum sanctum can be identified by angular epidermal
cells and its peculiar glandular and simple trichomes glandular trichomes with 8 celled head and simple
trichomes 3-5 celled with pointed apex (Figure 3a) while
in other species epidermal cells are wavy and different
type of trichomes. For example, two types of glandular
trichomes – unicellular, 18 – 28 m and 72 – 88 m with
globular head in O. basilicum (Figure 3b); two types of
trichomes - glandular trichomes with single basal cell but
simple and larger trichomes with multicellular bases in O.
canum (Figure 3c); single type of large sized glandular
trichome in O. gratissimum (Figure 3d).
a. O. sanctum
c. O. canum
b. O. basilicum
d. O. gratissimum
Figure 3. Leaf Surface anatomy of Rehan.
iv. ‘Zarnab’
The official drug consists of leaves of Abies spectabilis
Spach of the family - Pinaceae but leaves of Taxus
wallichiana Zucc. of the family – Taxaceae are being used
or sold as drug „Zarnab‟. The leaves of A. spectabilis are
linear, flattened with a prominent midrib and bifid apex
while in T. wallichiana these are linear lanceolate with
pointed apex. In A. spectabilis upper epidermis has
elongated rectangular epidermal cells, sinuous, thick
walled while lower surface has 7-9 rows of stomata in the
middle of the lamina. On the contrary, In T. wallichiana
upper epidermal cells are angular, hexagonal and thin
walled interrupted with trichomes and lower epidermis
has peculiar sunken stomata arranged in 6-7 rows in the
middle of the lamina (Rawat et al., 1996).
Int J Adv Pharmacy Med Bioallied Sci. 3, 1, 2015.
3. Planar chromatography
The official IUPAC term “planar chromatography”, covers
all chromatographic techniques using a planar open
stationary phase: thin-layer chromatography (TLC), highperformance
thin-layer
chromatography
(HPTLC),
ultrathin-layer chromatography (UTLC), overpressure
layer chromatography (OPLC) and preparative layer
chromatography (PLC) (Anonymous 2014). However,
TLC/HPTLC is the most powerful analytical version of this
form of chromatography due to its advantages which
include simple, cost-effective, versatile, precise sample
application, wide choice of stationary phases, multiple
sample handling, visual chromatogram, simultaneous
processing of standards, simultaneous scanning in
different light sources, disposable layer and usable in all
laboratories etc. The uses of this analytical technique in
quality evaluation of plant materials include fingerprint
profiling for the assessment of chemical constituents,
identification of adulterants/substitutes and quantitative
estimation of bio-markers in plant drugs.
Quality
Evaluation
&
Identification
of
adulterants/substitutes using TLC/HPTLC
In pharmacopoeias (i.e. Indian Pharmacopoeia; Unani
Pharmacopoeia of India & Ayurvedic Pharmacopoeia of
India) the Rf values of general TLC profiles have been
included as minimum requirement of quality standards.
However, TLC/HPTLC is being used extensively in the
recent years for fingerprinting of medicinal plants as
quality
control
markers,
identification
of
adulterants/substitutes and batch to batch consistency of
the products (Khatoon et al., 2005, 2010, 2011, 2014;
Singh et al., 2009; Gupta et al., 2011). Examples:
i. Bach
Bach consists of dried rhizome of Acorus calamus L.;
Fam. Araceae. Major constituent is asarone. Samples
were collected from Lucknow, Dehradun and also
procured from Delhi market. Applied the test and
reference solutions (10l) at 4 different tracks on a
precoated silica gel G plate 60 F254 of uniform thickness
(0.2mm). The plate was developed in the solvent systemtoluene : ethyl acetate (9 : 1), to a distance of 9 cm and
scanned densitometrically at 254 nm. Asaron (at Rf 0.56)
ranges from 0.54 – 1.16 % (Govindarajan et al., 2003).
ii. ‘Khulanjan’
Khulanjan consists of dried rhizome of Alpinia galanga
Willd. (Fam. Zingiberaceae). Another species A.
officinarum is also known as „Khulanjan‟. The samples of
A. galanga were collected from Dehradun, Jammu &
Tirunelveli. TLC of the methanolic extract on precoated
silica gel G plates using toluene : ethyl acetate :
methanol (80 : 20 : 0.4) shows under UV (366 nm) blue
fluorescent zones of yellow, green and blue at Rf. 0.15,
0.25, 0.69 respectively. After derivatization with
anisaldehyde- sulphuric acid reagent 10 minutes at 1200C,
spots appear at Rf. 0.15 (greyish green), 0.35 (violet),
0.48 (greyish green), 0.63 (greyish green), 0.69 (green)
and 0.91 (violet). The TLC profile of A. officinarum
www.biomedjournal.com
61
62
Khatoon | Quality control tools for Unani drugs
showed additional spot at Rf 0.31 (yellow) under UV 366
nm and at Rf 0.31 (dark blue), 0.51 (orange), 0.57, 0.59
(both dark blue), 0.63, 0.78 (both pink) and 0.90 (brown)
after derivatization (Figure 4).
Figure 5. TLC fingerprint of Khulanjan.Samples- AO- A.
officinarum 1-3. A. galanga. 1.Dehradun; 2. Jammu; 3.
Tirunelveli.
iii. Resha Khatmi
Roots of Althaea officinalis are official drug „Resha
Khatmi‟ but the roots of Alcea rosea syn. Althaea rosea
are being mixed with this drug. Sample of Althaea
officinalis was collected from Srinagar while roots of
Alcea rosea were collected from Lucknow. TLC of the
methanolic extract on precoated silica gel G plates (0.2
mm thick) using toluene : ethyl acetate : methanol (80 :
20 : 0.05) shows common as well as differentiating bands
i.e. at Rf 0.24 and 0.73 (both blue) under UV 366 nm and
blackish blue at Rf 0.59 and 0.69 under visible light after
derivatization in both the species. Additional orange
bands at Rf 0.11, 0.13, 0.37 and 0.41 were only visible in
Althaea officinalis (Khatoon et al., 2008a).
iv. Phyllanthus sps.
P. amarus, P. fraternus Webster. and P. maderaspatensis
can be identified by their TLC profile even in extract
form. TLC fingerprints were developed in a solvent
system – toluene : ethyl acetate (8.5 : 1.5) and visualized
the plate after derivatization with anisaldehyde sulphuric
acid reagent. TLC finger print profiles of P. amarus and
P. fraternus were almost similar. However, an additional
band of bright blue colour at Rf 0.90 was observed only in
P. fraternus and the bio-markers- phyllanthin and
hypophyllanthin were detected only in P. amarus. In P.
maderaspatensis only two bands at Rf 0.28 and 0.38 were
observed (Khatoon et al., 2006b).
iv.
Terminallia sps.
Barks of Terminalia species are being used as cardiac
tonic, diuretic. These are widely used as important
timber yielding plants and are rich source of tannins and
dyes. The stem bark (waste of timber industry) can be
exploited for important polyphenols- ellagic acid and
gallic acid, which have potent antioxidant potentials.
HPTLC markers and a method for quantification of ellagic
acid and gallic acid in seven Terminalia species viz T.
arjuna (Roxb.) Wight & Arn, T. bellirica Roxb. , T.
bialata Steud., T. catappa Linn., T. chebula Retz.., T.
manni King and T. tomentosa W. & A. Prodr., have been
developed. The study showed that common as well as
distinguishing bands were observed for all seven
Terminalia species in UV light at λ= 254 nm and 366 nm
and after derivatization with anisaldehyde sulphuric acid
reagent. The percentage of ellagic acid and gallic acid
Int J Adv Pharmacy Med Bioallied Sci. 3, 1, 2015.
varies from species to species. The maximum
concentration of ellagic acid was found in T. chebula and
that of gallic acid in T. catappa i e. 2.69% and 1.04%
respectively (Khatoon et al., 2008b).
vi.
Effect of microbial load on glycyrrhizin of
Glycyrrhiza glabra L.
Glycyrrhiza glabra, commonly known as „Asl-us-soos‟, is
an age old plant used to cure varieties of ailments from
simple cough to hepatitis more complex like Severe Acute
Respiratory Syndrome (SARS) and cancer. Microbial
contamination in the crude drug may occur through
handling by personnel. The presence of E. coli,
Salmonella spp. and moulds indicate poor quality. It was
observed that the concentration of glycyrrhizin (active
constituent) decreases by microbial contamination. The
TLC plate was developed in the solvent system
chloroform: acetic acid: methanol: water (6 : 3.2 : 1.2 :
0.8). The band of glycyrrhizin was observed at Rf 0.72
under UV 254 nm which converted purplish blue colour
after derivatization with anisaldehyde-sulphuric acid
reagent (Agarwal et al., 2014).
vii. Effect of seasons & dioecy on bio-markers of
Tinospora cordifolia (Thunb.) Miers
T. cordifolia (family-Menispermaceae) is a dioecious
creeper, commonly known as „Gilo‟. Male and female
plants of T. cordifolia were collected in the months of
January, April, June, August and October for studying
seasonal as well as dioecy variation. The HPTLC was
developed with chloroform: methanol: water (8: 2: 0.2)
as mobile phase. Densitometric scanning at 220 nm for
tinosporaside and at 320 nm for berberine was evaluated
as shown in Figure 5. The biomarkers- tinosporaside and
berberine reached in their highest concentration in
monsoon season and female samples showed significantly
higher concentration of aforesaid bio-markers. (Choudhry
et al. 2014).
10
1
Ber
Tin
10
1
Figure 5. HPTLC Densitometric fingerprint profile of
methanolic extracts of Tinospora cordifolia stem
collected in different seasons along with biomarkers. 1,
3, 5, 7, 9: male samples; 2, 4, 6, 8, 10: female samples
www.biomedjournal.com
Khatoon | Quality control tools for Unani drugs
(1-2: January; 3-4: April; 5-6: June; 7-8: August; 9-10:
October); Ber: Berberine; Tin: Tinosporaside.
CONCLUSIONS
WHO has developed a series of technical guidelines and
documents relating to the safety and quality assurance of
medicinal plants and herbal drugs. These include,
Guidelines on good agricultural and collection practices
(GACP) for medicinal plants (Anonymous 2003), Quality
control methods for medicinal plant (Anonymous 1998)
and Guidelines on good manufacturing practices (GMP)
for herbal medicines (Anonymous 2007). From the ongoing
studies it can be concluded that the macro-microscopy
and planar chromatography are the primary and
important tools for quality evaluation and identification
of adulterants/substitutes of Unani drugs. The
concentration of therapeutically active phytoconstituents
can easily be assessed by using TLC/HPTLC. These
parameters not only are utilized by students and
academicians but also by pharmaceutical companies for
proper quality of Unani drugs and batch to batch
consistency of their compound formulations.
ACKNOWLEDGEMENTS
Author is thankful to the Director, CSIR-National
Botanical Research Institute, Lucknow, India for providing
the facilities and Dr. Shanta Mehrotra (Ex-Head) & Dr.
AKS Rawat, Head Pharmacognosy and Ethnopharmacology
Division for encouragement throughout the work.
REFERENCES
Agarwal M, Rai V, Khatoon S, Mehrotra S. Effect of microbial
load on therapeutically active constituent glycyrrhizin of
Glycyrrhiza glabra L. Indian Journal of Traditional
Knowledge. 2014;13(2):319-324.
Aneesh TP, Hisham M, Sekhar MS, Madhu M, Deepa TV.
International market scenario of traditional Indian herbal
drugs - India declining. International Journal of Green
Pharmacy 2009;3:184-90.
Anonymous. The Wealth of India. Vol. X (Sp-W): Council of
Scientific and Industrial Research, New Delhi: Publications
and Information Directorate;1982:580-585.
Anonymous. WHO monographs on selected medicinal plants.
Rhizoma Rhei. Vol 1, World Health Organization Geneva;
1999. p. 231-240.
Anonymous. Quality control methods for medicinal plants.
World Health Organization, Geneva; 1998.
Anonymous. WHO guidelines on good agricultural and
collection practices (GACP) for medicinal plants. World
Health Organization, Geneva; 2003.
Anonymous. Guidelines on good manufacturing practices
(GMP) for herbal medicines. World Health Organization,
Geneva; 2007.
Anonymous. International Symposium on thin layer
chromatography; 2014. [http://www.hptlc.com] (Assessed
online January 2015)
Choudhary N, Singh S, Siddiqui MB, Khatoon S. Impact of
Seasons and Dioecy on Therapeutic Phytoconstituents of
Tinospora cordifolia, a Rasayana Drug. Biomed Research
International.
2014.
[http://dx.doi.org/10.1155/2014/902138]
(Assessed on
January 2015)
Int J Adv Pharmacy Med Bioallied Sci. 3, 1, 2015.
Ghani N. Khazainul Adviyah. Vol. I. Munshi Nawal Kishore,
Lucknow;1920. pp. 230-231.
Govindarajan R, Agnihotri AK, Rawat AKS, Khatoon S,
Mehrotra S. Pharmacognostic evaluation of an antioxidant
plant – Acorus calamus Linn. Natural Product Sciences. 2003;
9(4):264-269.
Gupta M, Bisht D, Khatoon S, Srivastava S, Rawat AKS.
Determination of Ursolic Acid a Biomarker in Different
Swertia Species through High Performance Thin Layer
Chromatography. Chinese Medicine. 2011;2(4):121-124.
Ibn-Al-Bitar. Al Jamia Li Mufrradat Al Adwiyyah Wa Al
Aghdhiya. Vol.1. Translated in Urdu by Yusuf M, Hassan SM.
Central Council for Research in Unani Medicine (CCRUM),
New Delhi; 1985.
Kale VS. Variable rates of primary and secondary metabolites
during different seasons and physiological stages in
Convolvulus, Datura and Withania. Asian Journal of
Experimental Biological Sciences. 2010: 50-53.
Khatoon S, Agnihotri AK, Singh N, Rawat AKS, Mehrotra S.
Comparative Pharmacognostic Evaluation of Althaea
officinalis and Alcea rosea root. Hamdard Medicus.
2008a;51(2):56-62.
Khatoon S, Rai V, Rawat AKS, Mehrotra S. Comparative
pharmacognostic studies of three Phyllanthus species.
Journal of Ethnopharmacology. 2006b;104:79-86.
Khatoon S, Rawat AKS, Mehrotra S. Commercial Crude Herbal
Drugs and Their Quality. Indian Journal of Development
Research & Social Action. 2006a;2(2):247-258.
Khatoon S, Singh H, Goel AK. Use of HPTLC to Establish the
Chemotype of a Parasitic Plant, Dendrophthoe falcata (Linn.
f.) Etting. (Loranthaceae), Growing on Different Substrates.
Journal of Planar Chromatography. 2011;24(1):60-65.
Khatoon S, Singh H, Rathi A, Ojha SK, Rawat AKS.
Standardization of an Ayurvedic Formulation- Kalyanavleha
and estimantion of Curcumin using HPTLC. Indian Journal of
Traditional Knowledge. 2014;13(3):535-542.
Khatoon S, Singh H, Singh K, Goel AK. TLC evaluation and
quantification of phenolic compounds in different parts of
Dendrophthoe falcata (Linn.f.) Etting, Journal of Planar
Chromatography. 2010;23(2)104-107.
Khatoon S, Singh N, Srivastava N, Rawat AKS, Mehrotra S.
Chemical Evaluation of Seven Terminalia Species and
Quantification of Important Polyphenols using HPTLC.
Journal of Planar Chromatography. 2008b;21(3):167–171.
Khatoon S, Srivastava M, Rawat AKS, Mehrotra S. HPTLC
Method for Chemical Standardization of Sida Species and
Estimation of the Alkaloid Ephedrine. Journal of Planar
Chromatography. 2005;18:364-367.
Khatoon S. Pharmacognostic & Chemotaxonomic studies of
Indian „Ratanjot‟ and their authentication in crude drug
markets of India. PhD Thesis. Aligarh Muslim University,
Aligarh; 1991.
Khatoon S, Mehrotra S. Bark Drugs. Vol. 1. CSIR – National
Institute of Science Communication and Industrial Research,
New Delhi; 2009.
Kroymann J. Natural diversity and adaptation in plant
secondary metabolism. Current Opinion in Plant Biology.
2011;14(3):246–251.
Kumar A, Kaul MM, Bhan MK, Khanna PK, Suri KA.
Morphological and chemical variation in 25 collections of the
Indian medicinal plant Withania somnifera. (L.) Dunal
(Solanaceae). Genetic Resources and Crop Evolution.
2007;45:665-660.
www.biomedjournal.com
63
Khatoon | Quality control tools for Unani drugs
Kunle OF, Egharevba HO, Ahmadu PO. Standardization of
herbal medicines - A review International Journal of
Biodiversity and Conservation. 2012;4(3):101-112.
Tuli R and Sangwan RS, Editors. Ashwagandha (Withania
somnifera) A model Indian Medicinal Plant. CSIR- National
Botanical Research Institute, Lucknow; 2009.
Li SP, Zhao J, Yang B. Strategies for quality control of
Chinese medicines. Journal of
Pharmaceutical and
Biomedical Analysis. 2011;55: 802-809.
Zhong Linda LD, Hu Dong-Dong, Xu Hong-Xi, Han Quan-Bin,
Bian Zhao-Xiang. Efficacy-Driven Quality Control Platform for
Chinese Herbal Medicine. Pharmceutica Analytica Acta
2014;5:291. [http://dx.doi.org/10.4172/2153-2435.100029]
(Assessed on February 2015).
Mehrotra S, Rawat AKS, Khatoon S, Pushpangadan P.
Adulteration and substitution in Herbal Drugs – A review. In:
Majumdar DK, Govil JN, Singh VK, Editors. Recent progress in
medicinal plants. SCITECH Publications, USA; 2001;8:177191.
Mehrotra S, Rawat AKS, Shome U. Standardization and
quality evaluation of „Banafsha‟. Natural product sciences.
1998;4(1):15-22.
Mehrotra S. Standardization and quality control. In: Post
Harvest technology of Medicinal and Aromatic plants. Tewari
SK, Pushpangadan P, Editors. National Botanical Research
Institute; 2003. p. 112-120.
Mitra RL, Jain SK. Concept of Phyllanthus niruri
(Euphorbiaceae) in Indian floras. Bulletin of Botanical Survey
of India. 1985;27:161–176.
Nadkarni KM. Indian Materia Medica: Popular Prakashan Pvt.
Ltd. India; 1996. P. 142.
Nizami Q, Jafri MA. Unani drug Jadwar (Delphinium
denudatum Wall.) – A review. Indian Journal of Traditional
Knowledge. 2006;5(4):463-67.
Pavarini DP, Pavarini SP, Niehues M, Lopes NP. Exogenous
influences on plant secondary metabolite levels. Animal
Feed Science and Technology. 2012;176,(1–4):5–16.
Prakash O, Jyoti AK, Pavan K, Manna NK. Adulteration and
Substitution in Indian Medicinal Plants: An Overview. Journal
of Medicinal Plants Studies. 2013;1(4):127-132.
Rawat AKS, Mehrotra S, Shome U. Comparative
pharmacognostic studies on the leaves of Abies spectabilis
and
Taxus
wallichiana.
International
Journal
of
Pharmacognosy. 1996;34 (5):378-383.
Rowson JM. Diurnal variation of glycosidal content of leaves
of Digitalis purpurea L. Pharmaceutical Acta Helvetiae.
1961; 36:69-73.
Sagar PK. Adulteration and substitution in endangered, ASU
herbal medicinal plants of India, their legal status, scientific
screening
of
active
phytochemical
constituents.
International Journal of Pharmaceutical Sciences and
Research. 2014;5(9):4023-4039.
Sharma PV. Caraka Samhita. 7th Edition, Chaukhambha
Orientalia, Varanasi; 2001.
Caraka-Saṃhitā: Agniveśa's Treatise Refined and annotated
by Caraka and Redacted by Dṛḍhabala (text with English
translation). Chaukhambha Orientalia, Varanasi; 1994.
Singh N, Khatoon S, Srivastava N, Rawat AKS, Mehrotra S.
Qualitative and Quantitative Standardization of Myrica
esculenta Buch.-Ham. Stem Bark through HPTLC, Journal of
Planar Chromatography. 2009;22(4):287-291.
Singh S, Khatoon S, Singh H, Behera SK, Khare PB, Rawat
AKS. A report on pharmacognostical evaluation of four
Adiantum species, Pteridophyta, for their authentication and
quality control, Revista Brasileira de Farmacognosia
(Brazilian Journal of Pharmacognosy). 2013;23(2):207-216.
Tewari RC, Chaubey S, Bal N, Kour G. Guidelines for
collection of raw drugs and characteristics of collected
material W.S.R. to Ayurveda. Unique Journal of Ayurvedic
and Herbal Medicines. 2014;2(3):4-7.
Int J Adv Pharmacy Med Bioallied Sci. 3, 1, 2015.
www.biomedjournal.com
64