Pharmacognostic evaluation of Withania coagulans Dunal (Solanaceae) -an important ethnomedicinal plant

Abstract

Withania coagulans Dunal, belonging to the family Solanaceae, is a small bushy shrub which is widely spread in South Asia. The plant is commonly known as 'Indian cheese maker' or 'paneer dodi' due to its milk coagulating characteristics of the fruits. In traditional system of medicine, different parts of plant especially fruits are used as magic healer of various diseases. In the present work, pharmacognostical studies of fruits and seeds are carried out for authentication of drug plant. Physico-chemical and phyto-chemical screening of drug material are done for determination of quality/purity of crude drug and for detection of plant constituents respectively. The plant is characterized by shrubby habit with dioecious and polygamous flowers; fruits (berries) enclosed in persistent leathery calyx; seeds ear-shaped, with fruity smell. Fruit pedicel with branched and unbranched trichomes, massive collenchymatous cortex, intra-xylary phloem and hollow pith; calyx with spongy parenchyma; pericarp with exocarp, mesocarp and endocarp; seeds with highly lignified sclerenchyma cells and strongly thickened endosperm. The plant is rich in alkaloids, esterase, carbohydrates, steroids, phenolic compounds, tannins, free amino acids and organic acids.

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ISSN: 2229-3469 (Print); ISSN: 2231-024X (Online)
Received: 07-06-2014, Revised: 20-12-2014, Accepted: 27-12-2014
Full Length Article
Pharmacognostic evaluation of Withania coagulans Dunal
(Solanaceae) - an important ethnomedicinal plant
Debasmita Dutta Pramanick* and S. K. Srivastava
Botanical Survey of India, Northern Regional Centre, Dehradun-248 195
*debasmita_bsi@yahoo.com
ABSTRACT
Withania coagulans Dunal, belonging to the family Solanaceae, is a small bushy shrub which is
widely spread in South Asia. The plant is commonly known as ‘Indian cheese maker’ or ‘paneer
dodi’ due to its milk coagulating characteristics of the fruits. In traditional system of medicine,
different parts of plant especially fruits are used as magic healer of various diseases. In the
present work, pharmacognostical studies of fruits and seeds are carried out for authentication of
drug plant. Physico-chemical and phyto-chemical screening of drug material are done for
determination of quality/purity of crude drug and for detection of plant constituents respectively.
The plant is characterized by shrubby habit with dioecious and polygamous flowers; fruits
(berries) enclosed in persistent leathery calyx; seeds ear-shaped, with fruity smell. Fruit pedicel
with branched and unbranched trichomes, massive collenchymatous cortex, intra-xylary phloem
and hollow pith; calyx with spongy parenchyma; pericarp with exocarp, mesocarp and endocarp;
seeds with highly lignified sclerenchyma cells and strongly thickened endosperm. The plant is rich
in alkaloids, esterase, carbohydrates, steroids, phenolic compounds, tannins, free amino acids
and organic acids.
Key Words: Indian cheese maker, Pharmacognostic evaluation, Withania coagulans, conservation.
INTRODUCTION
In Indian systems of medicine, a large
number of herbal and herbo-mineral drugs are in
use for ailment of various types of diseases of
human since long (Brekhman and Dardimov, 1969).
The traditional medicines are increasingly solicited
through the traditional practitioners and herbalists
in the treatment of different diseases. Currently
more than 80% of the world population depends
on plant derived health care products for daily
regime because of no side effect (Hassan et al.,
2009; Gangadhar et al., 2012). Withania coagulans
Dunal (Solanaceae), commonly known as
‘vegetable rennet’ or ‘Indian cheese maker’, is well
known in Indian medicine system due to its
multifarous medicinal and ethno-pharmacological
uses. It is distributed in the east of the
Mediterranean region and extends to South Asia
(Gupta, 2012). In India, the plant is common in
drier parts of Punjab, Haryana, Gujarat and
Rajasthan and known by different local names viz.,
‘Akri’ or ‘Puni-ke-bij’ in Hindi, ‘Tukhme-kaknaje-
hidi’ in Persian, ‘Spiubajja’ in Afghan, ‘Khamjira’ in
Punjabi, ‘Punir band’ or ‘Punir-ja-fota’ in Sindhi
(Naz et al., 2009). In folk medicine, different parts
of the plant, especially fruits are considered as
magic healer (Gupta and Keshari, 2013). The fruits
are reported to be sedative, emetic and diuretic.
They are effective in chronic liver disorder,
dyspepsia, flatulent coli and other intestinal
infections, asthma, biliousness and strangury
(Gupta, 2012; Gupta and Keshari, 2013). In some
parts of the Indo-Pak sub-continent, the berries are
used as a blood purifier.

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Debasmita Dutta Pramanick and S K Srivastava
The twigs are chewed for cleaning teeth
and the smoke of the plant is inhaled for relief of
toothache (Kirthikar and Basu, 1933). In North-
western parts of India, traditional practitioners use
dry fruits of this species for the treatment of
diabetic patients though its anti-hyperglycemic
activity has not been evaluated systematically.
Being a highly valued ethnomedicinal plant of drier
parts of the country, skill-less and unscientific
uprooting of the whole plant is being practiced
with upcoming threats. Thus, ex-situ and in-situ
conservation is needed for protection of this plant
in near future (Nautiyal, 2011; Rathore et al.,
2012).
MATERIALS AND METHODS
a) Macroscopical and Microscopical studies:
The fruits of Withania coagulans (with
persistent calyx and pedicel), commonly known as
‘paneer phul’ or ‘paneer dodi’, were purchased
from the local drug market of Kolkata and
identified by Pharmacognosy Department, NRIADD,
Kolkata. After proper identification with the help of
standard flora, the voucher specimen
(WCDP00011) has been deposited in the above
mentioned department, Kolkata. The fruits have
been examined macroscopically with reference to
its colour, shape, size, odour and taste. Hand
sections of the pedicel, calyx, pericarp and seeds of
the water soaked fruits have been done, stained
and mounted following standard method
(Johansen, 1940). For phyto-chemical studies fruits
have been dried in shade and were grinded to
coarse powder (Johansen, 1940; Kay, 1938; Trease
and Evans, 1983).
b) Physico-chemical studies:
Physico-chemical parameters of the
powdered drug such as total ash, acid insoluble
ash, water soluble extractive value and alcohol
soluble extractive value have been determined
according to the procedures mentioned in
Pharmacopoeia of India (1996).
c) Phyto-chemical studies:
Phyto-chemical studies have been carried
out by preparing 50% ethanolic extract of fruit and
has been subjected to different chemical tests for
identification of various bioactive compounds
(Kokate, 2001).
d) Fluorescence characteristics:
For study of fluorescence characteristics,
air dried coarse powder of fruits has been
examined under ultraviolet light according to the
method described by Chase and Pratt (1949) and
Kokoski et al. (1958).
RESULTS AND DISCUSSION
Botanical description:
Stout, greyish-white, small shrub, 30-90 cm
tall. Leaves 2.5-7.5 cm x 1.0-1.5 cm, lanceolate
oblong, sometimes ovate, obtuse, entire, narrowed
at the base, densely covered with minute, gray,
stellated tomentum; shortly petiolate.
Inflorescence axillary cymose. Flowers yellow, 7-12
mm across, dioecious and polygamous with
leathery calyx. Berries globose, red or brown,
smooth, covered enclosed in leathery calyx. Seeds
dark brown, ear shaped, glabrous, pulp brown,
having sharp fruity smell.
Fig. 1: Withania coagulans Dunal: A. Fruits; B. Powder of fruits (Churna)

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Table 1: Physico-chemical observations (% w/w )
Test
Observation 1
Observation 2
Observation 3
Total ash value
19.9
19.2
19.25
Acid insoluble ash
13.9
12.55
13.7
Water soluble extractive value
29.7
29.0
27.1
Alcohol soluble extractive value
6.8
6.5
6.0
Table 2: Phyto-chemical screening of 50% extract of fruits
Sr. No.
Tests
50% ethanolic extract
1.
Alkaloids
a. Dragendorff’s test
b. Wagner’s test
c. Mayer’s test
d. Hager’s test
+ve
+ve
+ve
+ve
2.
Carbohydrate
a. Molisch test
b. Fehling's Test
+ve
+ve
c. Benedict's Test
+ve
3.
Protein
a. Biuret Test
b. Xanthoprotein Test
c. Lead Acetate Test
+ve
+ve
+ve
4.
Amino acids
a. Ninhydrin test
+ve
5.
Glycoside
a. Baljet Test
b. Keller Kiliani Test
-ve
-ve
6.
Saponin test
a. Foam test
-ve
7.
Tannins and phenol compounds
+ve
8.
Essential oils
a. spot test
+ve
Table: 3- Fluorescence characteristics
Treatment with reagent
Fluorescence under long UV(366nm)
Powder as such
Brown
Powder treated with nitrocellulose in amyl acetate
Dark brown
Powder treated with 1N NaOH in methanol
Reddish green
Powder treated with 1N NaOH in methanol, dried
and mounted in nitrocellulose in amyl acetate
Dark green
Powder treated with 1N Hcl
Light green

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Debasmita Dutta Pramanick and S. K. Srivastava
Fig. 2. Withania coagulans Dunal: A. Transverse section of pedicel; B. Transverse section of calyx; C.
Transverse section of pericarp; D. Transverse section of seed.

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Fig. 3. Withania coagulans Dunal: Powder characteristics of fruits. A. Trichomes; B. Pericarp in surface
view; C. Upper epidermis of calyx; D. Fibre; E. Endosperm cells with aleurone grains; F. Portion of
cotyledons.

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Debasmita Dutta Pramanick and S. K. Srivastava
Macroscopical description:
The fruits are superior, reddish-yellow to
brown in colour, indehiscent, many-seeded berry,
pedicellate, round to globose, 6-9 mm in diam.,
enclosed within leathery persistent calyx, scurfy-
puberulous outside. The pedicel persistent, 3-7 mm
long. Pericarp smooth. Seeds 2.5-3 mm diameter,
ear-shaped, glabrous. The fruits have an indistinct
odour with a slight bitter taste [Fig. 1].
Microscopial description:
The transverse section of the pedicel shows
a single layer of epidermis composing of tabular
cells, covered with a large number of branched and
unbranched trichomes, followed by cortex
composing of 5-10 layers of collenchymatous cells.
The pericycle shows the presence of pericyclic
fibres with intervening parenchymatous cells. The
central region consisting of a narrow band of
phloem encircling the xylem beneath which is a
ring of intra-xylary phloem. The centre most region
is consisting of hollow pith surrounded by
parenchymatous cells with a few thick-walled
lignified fibres towards phloem [Fig.2(A)].
The transverse section of calyx exhibit a
single layer of thin-walled cells in both upper and
lower epidermis with a few branched and
unicellular covering trichomes present in upper
epidermis. The mesophyll is represented by spongy
parenchyma traversed by a number of small veins
covered with bundle sheath cells composed of thin-
walled parenchymatous cells [Fig. 2(B)].
The transverse section of pericarp (fruit
wall) shows the presence of exocarp which consists
of a single layer of cells while mesocarp shows a
wide zone of parenchymatous cells with strong
cellulosic thickening. The endocarp is consists of
single layer of cells [Fig.2(C)].
The transverse section of seed shows a
single layer of epidermis followed by a layered of
flattened thin-walled sub-epidermal cells. Beneath
the sub-epidermis there is a layer of highly lignified
sclerenchyma cells with narrow lumen. The inner
epidermis of the seed coat comprising of 1-3 layer
of thin-walled parenchymatous cells which at
places are collapsed. The endosperm is
represented by cells showing strong cellulosic
thickening filled with aleurone grains. The
cotyledon shows thin-walled radially elongated
cells enclosing a wide zone of round to oval to
polyhedral parenchymatous cells [Fig. 2(D)].
The powder characteristics of the fruits of
Withania coagulans has been described in [Fig. 3]
which shows (a) a large number of branched and
unbranched trichomes, (b) fragments of pericarp in
surface view, (c) epidermal cells of calyx with
unicellular covering trichomes, (d) asepted fibres,
(e) endosperm cells with aleurone grains and (f) a
large number of cotyledonary cells.
Physico-chemical observations:
Ash value help in determining the quality
and purity of crude drugs while extractive values
help in evaluating the constituens of crude drugs.
Various Physico-chemical Standards such as total
ash, acid insoluble ash, water soluble extractive
value, alcohol soluble extractive value have been
recorded (in triplicate) [Table 1].
Phyto-chemical screening:
The plant extract has been subjected to
preliminary phytochemical screening for the
detection of various plant constituents present in
the raw drug. The test results have been recorded.
[Table 2].
Fluorescence characteristics:
Fluorescence characters helps in fulfilling
the inadequacy of physical and chemical methods
for identification of plant drugs. Fluorescent
analysis of powdered drug on treatment with
different reagents has been studied under
ultraviolet light and the observations are presented
[Table 3].
CONCLUSION
The macroscopic and microscopic
evaluations of any plant drug are considered to be
the preliminary steps for establishing their quality
control profile. According to WHO, botanical
standards should be proposed as a protocol for the
diagnosis of the herbal drugs.
Physico-chemical standards, such as total
ash value help us in sophistication of drugs i.e. of
determining both physiological ash of plant tissues
and non-physiological ash of extraneous matters
like sand and soil; where as acid insoluble ash
detects presence of the heavy metals in the earthy
matter in the drugs. Extractive values (both water
soluble and alcohol soluble) help us in determining
the amount of active constituents and the above
mentioned tests are done on plant materials for
which still now no suitable chemical or biological
assay exists.

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In folk medicine, different parts of the plant
especially fruit (berry) are used as magic healer of
various diseases. Fruits of the plant have a milk
coagulating characteristics that is attributed as an
enzymatic charisma of the plant (Naz, 2002). The
milk coagulating property of the berries is due to
the presence of an enzyme, esterase. Most of the
activities of Withania coagulans is due to presence
of active constituent, ‘withanolides’ which are a
group of steroidal lactones with an ergostane
skeleton (Glotter, 1991). Withanolides which are
named after the name of the source plant Withania
species, are generally defined as C-28 steroidal
lactones. Modifications either of the carboxylic
skeleton or of the side chains result in many novel
structural variants of withanolides or ergostane-
type steroids (Cardenas et al., 1994). As
withanolides have been reported to posses anti-
tumorous, anti-angiogenic, chemopreventive and
inflammatory activities thus may represent useful
leads for development of potential anti-cancer
drug. In addition, withacoagin, coagulan,
withasomidienone, withaferin, 3-β-hydroxy-2,3-
dihydro-withanolide E, free amino acids, essential
oil etc. have been isolated from the berries as well
as aerial parts of the plant.
The natural habitat of W. coagulans is
destructed day by day by increasing rate of
agricultural practices, mining, industrialization,
reduction of forest cover and population explosion
throughout the country. In addition, global
warming, climate change and political partition of
India and Pakistan also had great impact on habitat
loss of the plant in India.However, some ex-situ and
effective micropropagation techniques have
already been implemented by Arid Zone Regional
Centre, Botanical Survey of India, some research
institutes, Universities and NGOs.
The major aim of present study is to
protect adulteration of crude drugs of plant origin
and give emphasis on conservation of W.
coagulans.The results have been obtained from the
present study may play an important role in setting
particular standards for the plant specimen. The
pharmacognostical, botanical, chemical and
economic characterizations also prove beneficial in
proper identification of plant drugs. Thus with the
help of these parameters the effect of adulteration
of fruits of this species can be minimized. Thus the
future research works with authenticated and
properly identified specimens will be of great use
(Prasad et al, 2010; Gupta, 2012; Salwaan et al.,
2012).
ACKNOWLEDGEMENT
The first author would like to thank to the Director
of NRIADD, Dept. of Ayush, Kolkata, for providing
multifarous facilities. Thanks are also expressed to
the research colleagues and technicians who
cooperated in carrying on the standardization
methods and helped in collecting the information
on this herb. Greatful acknowledgement is also
expressed to the Ministry of Health, Dept. of
Ayush, CCRAS for providing financial assistance.
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How to Cite this Article:
Debasmita Dutta Pramanick and SK Srivastava, 2015. Pharmacognostic evaluation of Withania coagulans
Dunal (Solanaceae) - an important ethnomedicinal plant. Bioscience Discovery, 6(1):06-13.