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PHARMACOGNOSTICAL EVALUATION OF THE FRUIT OF PLAKSHA -FICUS

LACOR Buch. Ham

Article · May 2014

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 4 | April 2014 | 165–174

ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal

Research Article

PHARMACOGNOSTICAL EVALUATION OF THE FRUIT OF PLAKSHA

- FICUS LACOR Buch. Ham.

Ahmad Ateeq1, Maurya Santosh Kumar2*, Seth Ankit3, Singh Anil Kumar4
1,2,3
Faculty of Ayurveda, Institute of Medical Sciences, Rajiv Gandhi South Campus, Banaras Hindu
University, Mirzapur–231001, Uttar Pradesh, India
4
Department of Dravyaguna, Faculty of Ayurveda, Institute of Medical sciences, Banaras Hindu University,
Varanasi–221005, Uttar Pradesh, India
*Corresponding Author: Email:– dravyapharma@gmail.com

Received: 06/03/2014; Revised: 28/03/2014; Accepted: 30/03/2014

ABSTRACT
Ficus lacor Buch. Ham. of the family Moraceae is a large glabrous tree, found throughout India.
It is well known for curing a variety of ailments such as gastric ulcer, Wound, dysentery, fever,
leucorrhoea and diarrhea. The present study was undertaken to evaluate the Pharmacognostical and
Phytochemical parameters of fruits of F. lacor. Macroscopically the fruit is green in color, depressed
and globose shaped, 1.2–4 cm in diameter and found astringent in taste. The results of microscopical
studies showed the presence of epidermis, lignified stone cells, collenchymatous tissue, etc. Powder
microscopy showed the presence of single or groups of lignified stone cells, collenchymatous cells
and lignified unicellular trichomes. The results of physiochemical parameters showed total ash 6.6%
w/w, water soluble ash 3.7% w/w, acid insoluble ash 0.4%w/w, Methanol soluble extractive 13.8%
w/w, chloroform extractive 11.9% w/w, water soluble extractive 35% w/w. The qualitative
evaluation of different extracts indicated the presence of carbohydrates, phenolic, triterpenoids,
protein and free amino acids compounds. Quantitative estimation by Spectrophotometry showed total
phenolic 19.4 mg/g (in gallic acid equivalent) and flavonoids 9.2 mg/g (in catechin equivalent). The
results from the present study reveal the standard monograph of the plants which will be essential
tools for the identification and authentication of F. lacor from other closely related species of Ficus.

KEYWORDS: Plaksha, Ficus lacor, pharmacognosy, phenolic, flavonoids

Cite this article:

Ahmad Ateeq, Maurya Santosh Kumar, Seth Ankit, Singh Anil Kumar (2014),
PHARMACOGNOSTICAL EVALUATION OF THE FRUIT OF PLAKSHA
- FICUS LACOR Buch. Ham., Global J Res. Med. Plants & Indigen. Med., Volume 3(4): 165–174

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 4 | April 2014 | 165–174
INTRODUCTION
Ficus lacor Buch. Ham. (syn. F. virens
Aiton) (Moraceae) is locally named as pakar in
Hindi. This plant is mentioned in different
Ayurvedic texts with different therapeutic
values and literatures. It is distributed
worldwide in Australia, South East Asia,
Burma, India, Bhutan, Indochina, Myanmar
and Nepal. It is found in India from Himachal
to Sikkim & in peninsular India up to an
altitude of 1000–2000 m (Khan et al., 2011a;
Chaudhary et al., 2012).
In Nepalese culture, tender buds and leaves
of F. lacor are used to make pickle and ripe
fruits are eaten fresh (Acharya and Acharya,
2010) while young shoots are eaten as
vegetable (Uprety et al., 2012). Stem bark is
used in gastric ulcer (Bajracharya et al., 1978;
Rai et al., 2004). Also it is useful in
gynecological disorders and gastric disorders
(Khan et al., 2011b; Shankar and Devalla,
2012). The bark of the plant is used for
expelling round worms, and for treatment of
leucorrhoea in Jatasankar Region of Girnar
Forest, Gujarat (Raval and Dhaduk, 2013).
Exudated from plant is used in typhoid and hay
fever (Paudyal, 2000), dysentery (Oli, 2001)
boils (Manandhar, 2002) and effective against
snake bite (Gomes, 2010). Aerial roots have
anti-arthritic property (Sindhu and Arora,
2013). Leaves of F. lacor has wound healing
(Pradeep, 2009), antioxidant activity (Ghimire,
2011) and used for treatment of various skin
problems (Gamble, 1967; Nadkarni and
Nadkarni, 1976). Decoction of buds is
considered to be good for ulcer and leucorrhoea
(Chopra et al., 1956), gargle in excessive
salivation condition (Malla, 1994), boils,
pimples and blisters (Manandhar, 1985). Fruits
of F. lacor Ham are used in diarrhea in
Pakistan (Khan et al., 2012). Seeds are tonic
and used in treatment of stomach disorder
(Bhatt, 1977). It is also used as an antibacterial,
antifungal (Swami and Bisht, 1996) and anti-
diabetic conditions (Chandira et al., 2010).
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MATERIALS AND METHODS
Chemicals and instruments
All the chemicals used were of laboratory
grade. Compound microscope, watch glass,
glass slides, cover slips, and other common
glassware’s were used in the present study.
Photographs were taken with using Magnus
Microscope Image Projection System (MIPS).
Folin-ciocalteu reagent (FCR), safranin, and
acetic acid were procured from Merck Pvt.
Ltd., India.
Preparation of Crude Extract
The samples were collected personally
during the month of October 2012 from its
natural habitat, Mughalsarai area of Chandauli
district (Uttar Pradesh). The sample was
taxonomically identified and authenticated by
Dr. Anil Kumar Singh, Professor (Dravyaguna),
Faculty of Ayurveda, Institute of Medical
Sciences, Banaras Hindu University Varanasi.
The specimen of the sample was deposited at
herbarium section of Department of
Dravyaguna, Faculty of Ayurveda, Banaras
Hindu University, Varanasi, India, for further
reference in future.
The fresh fruit of F. lacor were dried under
shade for two week, which were later
pulverized into coarse powder using
mechanical grinder, passed through 60 mesh
sieve and kept in a desiccator.
Pharmacognostical study
The study was carried out on the basis of
various pharmacognostical parameters viz.
macroscopic, microscopic, and fluorescence
behavior of powder. The macroscopy and
microscopy of the F. lacor fruit and its powder
were studied according to the method of Trease
and Evans, 2009. For the microscopical studies,
cross sections were prepared and stained as per
the procedure of Khandelwal, 2005. The
microscopic study was done according to the
method of Trease and Evans, 2009. Many herbs
show fluorescence behavior when exposed to
UV light and this can help in their
identification of crude drugs using the
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 4 | April 2014 | 165–174
fluorescence pattern. Fluorescence analysis was
carried out as per the standard procedures
(Lala, 1993). To study the fluorescence nature
of fruit powder, a pinch of powder was treated
with different chemical reagents viz.1N
hydrochloric acid, 1N sodium hydroxide, 1N
sodium hydroxide in methanol, picric acid, 1N
nitric acid, acetic acid, acetone, 50% sulphuric
acid, nitric acid in ammonia solution and
observed under day light, long UV (365 nm)
and short UV light (254 nm) (Laloo et al.,
2012).
Physico-chemical evaluation
Physico-chemical analysis was done as per
the standard methods and the WHO guidelines
on the quality control methods for medicinal
plants (WHO, 2002).
Preliminary phytochemical evaluation
The homogenous powdered sample was
extracted separately with different solvents
such as methanol, hexane, chloroform, ethyl
acetate and in water using cold maceration
process for 72 h. The extracts were filtered
through Whatman No. 1 filter paper and
concentrated using vacuum distillation to
generate the crude extracts of F. lacor fruit and
kept in a dessicator for further studies.
Preliminary phytochemical screening was
carried out by using standard procedures
(Khandelwal, 2005, Harborne, 1998).
Quantitative estimation
Determination of total phenolic content
The total phenolics content was determined
as per the methods of Folin-Ciocalteu assay
using standard gallic acid as the reference
control (Velioglu, 1998) with slight
modifications (Awah et al., 2012). Folin-
Ciocalteau reagent (FCR) consists of a yellow
acidic solution containing complex polymeric
ions formed from phosphomolybdic and
phosphor tungistic heteropoly acids.
Dissociation of a phenolic proton in a basic
medium leads to a phenolate anion which
reduces FCR forming a blue colored
molybdenum oxide whose color intensity is
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directly proportional to the phenolic contents
(Etim et al., 2013). Briefly, 1mL aqueous
extract was mixed with 1mL of FCR and
allowed to stand at 22ºC for 5 min. 4 ml
sodium carbonate was then added to the
mixture and the volume was made up to 10mL.
After 90 minutes, scanned at 760 nm in UV
spectroscopy and compare to standard
calibration curve. All the samples were
analysed in triplicate. Total phenol content
expressed in term of equivalent to gallic acid in
per g of dried extract.
Determination of Flavonoid content
Total flavonoid content was measured by
aluminium chloride colorimetric assay
(Zhishen et al., 1999) with slight modification
(Kural et al., 2011) using standard (+) catechin
as the positive control. An aliquots 1mL of
extracts was added to a 10 ml volumetric flask,
containing 4ml of distilled water. To the flask,
0.3 ml of 10 % NaNO2 was added. After 5 min,
0.3 ml of 10% AlCl3 was added. After 5 min, 2
ml of 1M NaOH was added and the volume
was made up to 10 ml with distilled water. The
solution was mixed well again and absorbance
was measured against a blank at 510 nm with
an UV-VIS spectrophotometer. The total
flavonoid content of the extract was expressed
as mg of (+) catechin equivalent (CE).
RESULTS AND DISCUSSION
Pharmacognostical evaluation
F. lacor fruits are synconus inflorescence
containing drupe fruits, having sub-globose
shape with 1.2–4 cm in diameter. The color of
the fruit changes from green (unripe fruit) to
greyish-brown or yellowish-brown (ripe fruit).
The outer surface of the fruit is smooth and
hard. Fruits have characteristic odour and
astringent taste (Fig 1a).
Transverse section showed that the fruit
was divided into three compartments viz.
epidermis, hypodermis and ground tissue.
Epidermis is thin-walled, single layered,
followed by a narrow zone of 2–5 layered
hypodermis which consist of round, oval,
rectangular, lignified stone cells with wide
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 4 | April 2014 | 165–174

lumen; rest of mesocarp very wide consisting
of oval to polygonal, collenchymatous cells
containing brownish contents; a few vascular
traces found scattered in this zone; inner zone
consisting of stone cells similar in shape and
size to those found scattered in outer zone.
Male and female flower attached to inner layer
of mesocarp (Fig 1b and Fig 1c).
Powder microscopy shows the fragments of
epidermal cells, single or group of lignified
stone cells, collenchymatous cells and lignified
unicellular trichomes, a few debris of male and
female flowers are also present (Fig 1d, Fig 1e
and Fig 1f).

Fig 1. Macro- & Microscopic plates of Ficus lacor

[a]: Ficus lacor fruit, [b-c]: Transverse section of fruit, [d]: Unicellular trichomes, [e]: lignified stone cells, [f]: vessels
elements. [UE - Upper Epidermis; CC- Cholenchymatous Cells; SC - Lignified Stone Cells; T - Trichomes; MR -
Medullary Rays]

Physico-chemical evaluation deterioration (WHO, 2002). The amount of

The physico-chemical parameters are
mainly used in evaluating the purity and quality
of the drug. The present study was conducted to
evaluate the various physicochemical
parameters on the fruits of F. lacor (Table 1).
The fruit was found to contain high amount of
moisture with a loss on drying value of 9.59 %
w/w. As per WHO the determination of loss on
drying (LOD) is important since LOD reveals
the percentage of moisture present in the drug.
Presence of high amount of moisture in any
drugs may facilitate the enzyme hydrolysis or
enhance the growth of microbes which leads to
foreign matter was found to be low with 0.20 %
w/w, since the plant was found to be less
contaminated with unwanted substance other
than the plant material itself. The total ash of
6.6% w/w, acid insoluble ash of 0.4% w/w and
water soluble ash of 3.7% w/w. Ash value of
drug gives an idea of the earthy matter or
inorganic composition or other impurities
present along with the drug (Laloo et al., 2013).
The foaming index of drug is less than 100,
indicating that the plant F. lacor is having less
quantity of saponin. The haemolytic index was
also found to be low with a hemolytic value of
10.5 units/g. In addition, the plant also showed

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low swelling index indicating the absence of Fluorescence analysis of powdered drug
gum, mucilage, pectin or hemicelluloses in the was studied in both UV and day light. The
sample. Table 1 showed the percentage powder showed different color fluorescence
extractive value of various extracts obtained when made to react with various chemical
from F. lacor fruit. The result reveals that the reagents which suggests that there might be a
aqueous extract showed the maximum certain phyto-constituent possessing
contribution of phyto-constituents. Extractive chromophore group present in the fruit (Table.
values give an idea about the chemical 2). Similar previous studies reported that the
constituents present in the drug as well as plant material have the capability to produce
useful in the determination of exhausted or fluorescence pattern when made to react with
adulterated drugs. various chemical reagents either of acidic or
basic media (Sahu et al, 2010, Laloo et al.,
2012).
Table. 1 Physicochemical evaluation
S.N Parameter Results
1. Foreign matter (% w/w) 0.20
2. Loss on drying (% w/w) 9.59
3. Ash Values
Total ash (% w/w) 6.6
Water soluble ash (% w/w) 3.7
Acid insoluble ash (% w/w) 0.4
4. Extractive values Extract colour
Water Saddle brown 35
Methanol Orange 13.8
Chloroform Yellow green 11.9
Ethyl Acetate Dark green 8.70
Hexane White 8.77
5. Foaming index Below 100
6. Hemolytic index 10.5 units/g
7. Swelling index Less than 1

Table. 2 Fluorescence analysis of fruit powder

Treatment Long U.V. (365 nm) Short U.V. (254 nm) Visible Light
NaOH + Methanol Pale green NF Dark sea green
NaOH + Water Yellow green NF Fire brick
HNO3 + Methanol Lime green NF Fire brick
HNO3 + Water Lime green Coral Navajo white
HCL + H2O Dark green NF White
HCL + Methanol Light green Dark olive green Coral
Iodine solution No color NF Saddle brown
NH3 Dark olive green NF Saddle brown
KOH Lime green NF Dark red
Glacial acetic acid Spring green NF Yellow green
Water No color NF Navajo white
NF: No Fluorescence

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Preliminary phytochemical evaluation layer chromatographic techniques. The TLC

Phytochemical screenings of all the
different extracts are shown in Table no. 3. The
results depicted that the phytochemicals present
in various extract from the fruit of F. lacor
were mostly carbohydrates, polyphenolics
(tannins, phenolics and flavonoids), alkaloids,
triterpenoid, proteins and amino acids. Whereas,
saponins, glycosides, and mucilage components
seems to be absent. Evaluation of crude drugs
on the basis of preliminary phytochemical
screening provides a brief idea about the
qualitative nature and distribution of active
phytoconstituents present in the fruit extract.
This can serve as essential diagnostic
parameters for the identification of crude drugs
(Prasad et al., 2013).
Results of preliminary phytochemical
screening were further confirmed by qualitative
chromatographic analysis incorporating thin
study was done in all the extracts obtained by
maceration. The results depicted that alkaloid
was present in only chloroform extract (Rf-
0.85); while, triterpenoids (Rf- 0.61–0.71)
components were found to be presence in all
the extracts except aqueous extract. Phenolics
compoents (Rf- 0.56–0.71) were found to be
present mostly in ethyl acetate, methanol and
aqueous extract. Moreover, methanolic and
aqueous extracts showed the presence of
carbohydrates (Rf- 0.83–0.91), amino acid (Rf-
0.83–0.91) and anthraquinone (Rf- 0.76–0.91)
(Fig 2). Analysis of phytochemicals on the
basis of TLC provides an essential knowledge
on the type and classes of phyto constituents,
since TLC operates on the separation of the
different classes of phytochemicals which can
be easily identified after derivatizing with
various classes of reagents (Sharma et al.,
2014).

Table. 3 Phytochemical screening of different extracts

Sr. Plant Constituents Test / Methanol Chloroform Ethyl acetate Aqueous Hexane
no. Reagent extract extract extract extract extract
1. ALKALOIDS
Dragendroff’s reagent + + − − −
2. CARBOHYDRATES
Molisch’s reagent + − + + −
Fehling solution + − − + −
Reducing sugar test + − − + −
3. TRITERPENOIDS
Salkowski test + + + − −
4. SAPONINS
Foam test − − − − −
Sodium bicarbonate test − − − − −
5. PHENOLIC COMPOUNDS & TANNINS
Ferric chloride solution + − − + −
Nitric acid test + − + − −
6. PROTEINS & AMINO ACIDS
Millon’s reagent + − − − −
Ninhydrin reagent + − − + −
7. FLAVONOIDS
Shinoda/Pew test + − − − −
Lead acetate test − − − + −
Zinc HCL test + − − − −
Alkaline reagent test − + − −
(+): Present; (−): Absent

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Fig 2. Thin layer Chromatographic study

[a]: Alkaloid under UV (365nm) light, [b]: Alkaloid under visible light, [c]: Triterpenoids under UV light, [d]:
Triterpenoids under visible light, [e]: Phenolics under UV light, [f]: Phenolics under visible light, [g]: Corbohydrates
under UV light, [h]: Carbohydrates under visible light, [i]: Amino acids under UV light, [j]: Amino acids under visible
light, [k]: Anthraquinone under UV light, [l]: Anthraquinone under visible light.
Abbreviation: MeE: Methanol extract; ChE: Chloroform extract; EaE: Ethyl acetate extract; AqE: Aqueous extract;
HeE: Hexane extract.

Quantitative estimation of phytoconstituents before conducting any isolation of active

biochemical markers since it gives a brief idea
From the quantitative estimations results, it about different classes of chemical
was observed that FLF contains maximum constituent’s present in the extract.
Total phenolic l19.4 mg/g (in gallic acid
equivalent). The estimated value of other CONCLUSION
polyphenolics constituent like flavonoids was
found to be 9.2 mg/g (in catechin equivalent). Herbal drugs used in various traditional
Estimation of phyto-constituents in the medicines need detailed ethno-pharmacological
extract/plant material is an important step exploration. This comprehensive

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pharmacognostical studies on the fruits of F.
lacor may substantiate as an essential data for
the identification of raw material and also used
to differentiate the plant from its allied species
and adulterants. This study would be the
leading path way of information for selection of
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Conflict of Interest: None Declared