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International Journal of Pharmacognosy and Phytochemical Research 2016; 8(5); 800-806

ISSN: 0975-4873
Research Article

Development of Quality Control Parameters for Standardization of

Leaves of Ficus Species
Shaikh A S1,2*, Mohammed Ibrahim3, Khan Mohib4
1
School of Pharmacy, AIKTC, Mumbai University, New Panvel-410206, India
2
Research scholar, JNTUH, Kukatpally, Telangan-50008, India
3
Nizam Institute of Pharmacy, Andhra Pradesh-508284, India
4
Oriental College of Pharmacy, Mumbai University, Navi Mumbai-400705, India

Available Online: 1st May, 2016

ABSTRACT
Ashvatha (Ficus religiosa Linn.), Nyagrodha (Ficus benghalensis Linn.), Udumbara (Ficus glomerata Roxb.) are few of
the reputed panchavlkal drugs of ayurveda found abundantly throughout India. Various parts of these plants are used as
antidiabetic, antidiarrheal, antistress, antioxidant, antiinflammatory, antiallergic, antiulcer, antimicrobial, hypolipidemic,
immunomodulatory and analgesic. In present study, a detailed comparative pharmacognostic study of leaves of Ficus
religiosa Linn., Ficus benghalensis Linn and Ficus glomerata Roxb. was carried out to develop quality control parameters
which could be useful as a reference for experimental study of these plants in future. The study includes macroscopy,
physicochemical evaluation, preliminary phytochemical screening, fluorescence analysis, UV Spectroscopic and thin
layer chromatographic evaluation.

Keywords: Ficus religiosa, Ficus benghalensis, Ficus glomerata, Macroscopic character, Physico-chemical study,
fluorescence analysis, UV Spectroscopic and chromatographic evaluation

INTRODUCTION
The herbal medicines is one of the oldest form of health Dr. A. S. Upadhye of Agharkar Research Institute, Pune,
care known to the mankind. It can be safely used if India. The leaves were dried in shade and powdered using
parameters such as authentication, quality control mixture grinder. The powder leaf material was preserved
standards and its efficacy and safety are maintained. In in air tight container for future use.
the view of the tremendous increase in use of herbal Plant extracts, chemicals and reagents
medicines, it is necessary to have standard parameters to The powdered plant material of leaf was successively
ensure its quality. Ficus religiosa, Ficus benghalensis, extracted with petroleum ether, benzene, chloroform,
Ficus glomerata are large and extensively growing trees ethanol and distilled water. All the extracts obtained are
found in Indian subcontinent known not only for its preserved in desiccator for future use. All the chemicals
religious but also for its medicinal value. The leaves, bark
and fruits of these plants are used as antiseptic, astringent,
haemostatic, antiinflammatory, anticancer, antioxidant
and also in the treatment of skin diseases, diarrhoea,
dysentery, vaginal disorders, ulcers, leucorrhoea,
deficient lactation, menorrhagia1. Although, the leaves of
these plants are important but very few studies reported
so far on comparative quality control parameters of these
theree plants from ficus species. Hence this study is
undertaken to develop comparative quality control
parameters of leaves of Ficus benghalensis (FB), Ficus
religiosa (FR) and Ficus glomerata (FG).

MATERIALS AND METHODS

Collection of plant material
The leaves of Ficus religiosa, Ficus benghalensis and
Ficus glomerata were collected from Sindhudurg district
of Maharashtra, India and identified with authentication Figure 1: Leaf and powder drug of FR, FB and FG
no. 14-235, 14-236 and 15-023 respectively by botanist

*Author for Correspondence

 Shaikh et al./ Development of Quality…

Table 1: Comparative macroscopic study of FB, FR and FG leaf

Leaf parts and FB FR FG
Characteristics
Duration Evergreen Evergreen Evergreen
Leaf arrangement Alternate Alternate Alternate
Petiole Color Size Brownish yellow (dried) Brownish yellow Dark brown (dried) 3-8 cm
3-5 cm (dried) 7-12 cm
Lamina Dark green (Fresh) Dark green (Fresh) Dark Olive green (Fresh)
Color Brownish yellow (dried) brown (dried) Light green (dried)
Shape Ovate-cordate Cordate with extended drip Ovate- Lanceolate
tip
Size Length -15-25 cm Length -10-15 cm Length - 8-17cm
Width - 7-18 cm Width -8-12 cm Width - 3.5-7 cm
Nature or composition Simple Simple Simple
Incision Absent Absent Absent
Venation Pinnate-Reticulate Pinnate-Reticulate Reticulate
Margin Entire Entire and slightly Entire
undulate (wavy)
Apex Mucronate Caudate- Acuminate Acuminate - Obtuse
Base Cordate-rounded Cordate Cuneate
Surface
Adaxia Pubescent Glabrous, Shiny Pubescent, glabrous
Abaxial Coriaceous Coriaceous Glabrous
Texture Thick and leathery, Thin, papery, smooth Thin,Papery, Smooth,
medium course texture Glaucouse
Taste Not significant Not significant Slightly acrid
Odour Odorless Odorless Odorless

Table 2: Comparative study of physical parameters of powdered leaf of of FB, FR and FG

Sr. Parameters FB FR FG
No
1 Total ash (%w/w) 13.5 14 14.5
2 Water soluble Ash (%w/w/) 2 6 2
3 Acid insoluble ash (%w/w/) 5 7 8
4 Water soluble extractive value (%w/w/) 21.2 19.6 18
5 Alcohol soluble extractive value (%w/w/) 9.2 2.8 8
6 Loss On drying (%w/w/) 5.55 3.05 4.48

Table 3: Comparative study of percentage yield for different extracts of FB, FR and FG
Sr. No Solvent Percentage yield
FB FR FG
1 Pet. ether 3.325 2.676 2.125
2 Benzene 1.075 1.595 0.675
3 Chloroform 0.7 0.925 0.85
4 Ethanol 1.975 3.55 2.371
5 Water - 12.775 7.447

and reagents used in this study are of analytical grade. was prepared by pouring silica gel on glass plate and
Development of standard analytical parameters: activated by heating at 110 0 c for 30 min. The sample
Macroscopic evaluation and physical parameters such as was spotted on the plate and the plate was kept in the
ash value, extractive value, loss on drying and percentage solvent system until it reaches to 3/4th of the plate. The
yield were performed according to the standard official spots are detected under long uv at 365 nm and short uv
method2. Preliminary phytochemical analysis of extracts at 254 nm and Rf value were calculated.
was carried out as per the standard method 3. UV
spectroscopic analysis of crude extracts was carried out RESULTS AND DISCUSSION
using Jasco UV Spectrophotometer in the range of 200 to Quality control of plants materials used in traditional
500 nm. Thin layer chromatography analysis is carried system of medicine is important for the
out using silicagel GF-254 as adsorbent, and benzene: commercialization of new therapeutic agents. Adulterated
chloroform (ratio of 7:3) is used as a mobile phase4. Plate and substituted plants materials may produce various

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Table 4: Comparative fluorescence analysis of powdered leaf sample of FB, FR and FG

Treatment FB FR FG
UV 254 nm UV 366 nm UV 254 nm UV 366 nm UV 254 nm UV 366 nm
As such Brown Black Brown Black Brown Black
Methanol Greenish Reddish Light green Reddish Greenish Reddish brown
brown brown brown brown
10 % NAOH Light green Black Light green Black Greenish Black
black
Conc. HCL Light green Black Light green Black Blackish Black
green
Conc. HCL + Greenish Blackish Light green Blackish Light green Black
H2O black green brown
Conc. Nitric Green Black Green Black Greenish Black
acid white
Conc. Nitric Blackish Bluish black Light green Black Light green Black
acid + H2O brown
Conc Blackish Bluish black Blackish Black Black Black
Sulphuric green green
Acid
Chloroform Light brown Black Light green Light orange Light green Reddish brown

Pet Ether Light green Reddish Transparent Brownish Light green Black
brown black

Table 5: Comparative study of treatment of powdered leaf with different chemicals

S. No Treatment with chemicals FB FR FG
1 Drug + Conc. HCL Dark brown Light brown Brown
2 Drug + Conc. HNO3 Reddish brown Orange Brisk red
3 Drug + Conc. H2SO4 Black Reddish black Black
4 Drug + Glacial acetic acid Light brown Light green Greenish brown
5 Drug + Picric acid Brown Light orange Yellowish brown
6 Drug + 5 % NaOH Light brown Light brown Brown

Table 6: Comparative phytochemical analysis of petroleum ether (PE), benzene (BE), chloroform (CH), ethanol(ET)
and aqueous (AQ) extracts of the powdered leaf of FB, FR, FG
Sr Extract Alkaloi Amino Anthra Flavon Glycos Polyph Saponi Tanins Triterp Steroids
. s ds acids quinon oids ide enols ns enoids
N e
o.
1 FBPE + - - - + - + - + +
2 FRPE + - - + - - - + + -
3 FGPE + - - + + + + - + +
4 FBBE - + - + - + + - + +
5 FRBE + - - + - + + + - +
6 FGBE + - - + + + - - + -
7 FBCH - - - + - + - - + +
8 FRCH + - - + + + + + - +
9 FGCH + - - + + + + - - -
10 FBET + - - + + + + + + +
11 FRET + - - + + + + + - -
12 FGET - - - + + + - + - +
13 FBAQ + + - + + + + + + -
14 FRAQ + - - + + + - + + -
15 FGQ + - - + - + + - - -

health related issues when used by the patients. Therefore, identification and standardization of these plants material.
comparative study of macroscopic features and quality Macroscopic evaluation:
control parameters of three plants of ficus species Data of different aspects of macroscopic evaluation is
mentioned in the present work may be helpful for given in Table 1. Although the three varieties belong to

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Table 7: Comparative UV spectroscopic analysis of extracts of leaf of FB, FR and FG

Sr. No. Solvent λ max of 0.1 mg/ml extract in respective solvents
FB FR FG
1 Pet. ether 369.7 268.6 269.6
2 Benzene 414.5 415 414.6
3 Chloroform 235.5 414.8 272.5
4 Ethanol 207.8 267.8 267.7
5 Water 215 218.5 213.9

Table 8: Comparative TLC analysis of petroleum ether (PE), benzene (BE), chloroform (CH), ethanol (ET) and
aqueous (AQ) extracts of the powdered leaf of FB, FR, FG
Sr. Extracts UV 254 nm UV 365 nm
No. Total No Color & Rf Value for Spot No Total No Color & Rf Value for Spot No
of Spots 1,2, n.. of Spots 1,2, n..
1. FBPE 2 1. Greenish yellow, 0.0363 2 1. Orange, 0.090
2. Greenish yellow, 0.090 2. Light blue,0.127
2. FBBE - - 1 1. Light Orange, 0.0769
3. FBCH 1 1. Greenish yellow, 0.0363 2 1. Green, 0.0363
2. Orange, 0.0727
4. FBET - - 2 1. Orange, 0.145
2. Light blue, 0.181
5. FBWA - - - -
6. FRPE 2 1. Yellow, 0.0727 3 1.Orange, 0.0727
2. Yellow, 0.109 2.Blue, 0.163
3.Blue, 0.545
7. FRBE 1 1. Greenish yellow, 0.0909 2 1. Orange, 0.0909
2. Blue, 0.545
8. FRCH 1 1. Greenish yellow, 0.0909 2 1. Orange, 0.0909
2. Light Blue, 0.5272
9. FRET 1 1. Light green, 0.145 2 1. Light orange, 0.0545
2. Dark orange,0.145
10. FRWA - - - -
11. FGPE 2 1. Greenish yellow, 3 1. Orange, 0.0727
0.036 2. Yellow, 0.1636
2. Greenish yellow, 3. Orange, 0.2727
0.109
12. FGBE 2 1. Light yellow, 0.0363 2 1. Light orange,
2. Greenish yellow, 0.0363
0.109 2. Dark orange,
0.109
13. FGCH 1 1. Greenish yellow, 2 1. Orange, 0.0909
0.0909 2. Light blue, 0.527
14. FGET 1 1. Light green, 0.196 1 1. Orange, 0.2
15. FGWA - - - -

same species, they vary with each other in many respects; acuminate while in FG it is acuminate- abtuse.
FB and FR leaves are large in size, dark green whereas Leaf base: In FB base is caurdate- rounded and FR it is
FG leaves are smaller and dark olive green in color with cordate while FG leaf
following macroscopic features: shows cuneate base.
Shape: FB leaves are ovate and cordate and FR leaves are Surface: In both FB and FR abaxial surface is coriaceous
cordate with extended drip teep while FG leaves are ovate while in FG it is glabrous whereas upper adaxial surface
lanceolate in shape (Photo slide 1- A, B, C). is pubescent in FB, glabrous and shiney in FR and
Venation: Both FB and FR showed pinnate- reticulate pubescent and glabrous in FG.
while FG showed reticulate type of venation. Texture: In FB leaf it has thick leathery medium course
Margine: Both FB and FG showed entire margine while texture while FR and FG leaf showed thin, papery and
FR showed entire and slightly wavey called as undulate smooth texture where as FG have additional glaucouse
margine. texture.
Apex: All three leaves showed different type of apex. In Determination of physico-chemical parameters:
FB leaf apex is mucronate and FR it is Caudate- Ash value

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Figure 2: Thin layer chromatography of petroleum ether (PE), benzene (BE), chloroform (CH), ethanol (ET) and
aqueous (AQ) extracts of the powdered leaf of FB, FR, FG at 365 and 254 nm

Ash values reflect quality, purity and authenticity of crud acid insoluble ash of the leaf of three plants expressed as
drug and also it is an important quantitative standard 5. average % w/w for three samples is given in table 2. Total
The comparative results of total ash, water soluble ash, ash of leaves of all three plants were found to be more

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than water soluble ash and acid insoluble ash. Water plants in the range of 200 to 500 nm were determined by
soluble ash of both FB and FG was found to be very less using jasco UV Spectrophotometer. Extract of leaves of
than total ash, acid insoluble ash of the leaf. all three plants showed different lambda max which is
Extractive value given in table7. Determination of Lambda max is
Extractive value is an indicator of total soluble indicative of wavelength at which compound shows
components in particular solvents. The comparative result maximum absorption. Since all extracts of three plants
of water soluble and alcohol soluble extractive value showed different lambda max values which reflect that
expressed as % w/w for three samples is given in table 2. the extracts are different in their composition of
Water soluble extractive value of leaves of all three plants phytoconstituents present in it.
were found to be more than alcohol soluble extractive Thin layer chromatography
value which indicate presence of more amount of water Thin layer chromatography is a very good tool to monitor
soluble components in the plants. the progress of a reaction, identify compounds present in
Loss on drying a given test drug and for ensuring the purity of a
Loss on drying for leaves of FB, FR and FG were found substance10. Thin layer chromatography of different
to be 5.5 % w/w, 3.05 % w/w and 4.48 % w/w which extracts of leaves of all three plants was carried out using
indicate presence of less moisture content in the crud silica gel GF-254 as adsorbent and benzene: chloroform
drug (Table 2). Less moisture content could prevent in the ratio of 7:3 as a mobile phase. Different extracts
herbal drugs from the growth of bacteria, fungi or yeast showed blue and orange colored fluorescence at 365 nm
during the storage6. with different Rf value which indicate presence of
Percentage yield flavonoids in the extracts (Table 8).
Percentage yield is the measure of the efficiency of
solvents to extracts specific components from the powder CONCLUSION
drug material7. Comprehensive comparative percentage Ficus benghalensis Linn., Ficus religiosa Linn. and Ficus
yield of powder leaf sample of three plants of ficus glomerata Roxb. are few of the reputed panchavlkal
species is given in table 3. Maximum yield is observed in drugs of ayurveda. Various parts of these plants are used
water extract of FR and FG followed by ethanol extract of as antidiabetic, antidiarrheal, antistress, antioxidant,
FR. However, benzene extract of FG followed by antiinflammatory, antiallergic, antiulcer, antimicrobial,
chloroform extract of FB showed minimum percentage hypolipidemic, immunomodulatory and analgesic etc. In
yield. spite of these immense medicinal value, there is lack of
Fluorescence analysis of leaf powder research work on comparative quality control parameter
Fluorescence analysis of powdered leaf sample has been of these three plants. Our present research work on the
carried out in visible light, UV light at 254 nm and 365 leaves of Ficus benghalensis Linn., Ficus religiosa Linn.
nm. The powdered leaf sample of all three plants were and Ficus glomerata Roxb. is an attempt for providing a
treated with various organic solvents and solutions and set of data that can be used as a reference to differentiate
observed under Visible and UV light (Table 4). The crude these closely related plants in future.
drugs produces fluorescence because in presence of
different solvents phytoconstituents in the drugs are ACKNOWLEDGEMENT
converted into fluorescent derivatives8. Since The author wish to thanks Dr. Abdul Razak Honnutagi,
fluorescence produced by the individual powdered drug is Director, AIKTC and Dr. Rajani Shettigar, Dean, SoP,
the characteristic of phytoconstituents present in it, AIKTC for providing facilities to carry out this work and
fluorescent analysis could be one of the important Dr. Shariq Syed, Associate Professor, SoP, AIKTC for his
parameter for evaluation of quality of herbal drugs. valuable edits.
Behavior of leaf powdered materials towards chemical
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