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Phytochemical studies on selected medicinal plant Gymnema sylvestre.

Keywords:
Phytochemical, Anti-inflammatory, Antimicrobial, TLC, Physicochemical
analysis, Fluorescent analysis.
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Dates:
Received: 06 Apr 2012 Accepted: 19 Apr 2012 Published: 21 May 2012
Article Citation:
Gnana Sangeetha D and Jegadeesan M.


Phytochemical studies on selected medicinal plant Gymnema sylvestre.
Journal of Research in Plant Sciences (2012) 1: 077-082
Original Research
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Authors:
Gnana Sangeetha D
1
.
and Jegadeesan M
2
.




Institution:
1. Department of Chemistry,
PSNA College of
Engineering and Technology,
Dindigul, Tamil Nadu, India

2. Department of
Environmental and Herbal
Sciences, Tamil University,
Thanjavur, Tamil Nadu,
India.






Corresponding author:
Gnana Sangeetha D.











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Journal of Research in
Plant Sciences
An International Scientific
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077-082 | JRPS | 2012 | Vol 1 | No 1
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ABSTRACT:
Gymnema sylvestre R. (Br).(Asclepiadaceae) is a common medicinal plant
available in Tamil Nadu is a woody, climbing plant that grows in the tropical forest of
central and Southern India. It also happens to be a first-rate warrior against diabetes.
The plant is called Gud-mar (Gud-Jaggery, mar-kills) in India and well known for
masking sweet taste. It is reported to be effective against many chronic diseases is
screened for its phytochemical content, microbial activity and anti-inflammatory
activity. Extracts (Alcoholic, aqueous, acetone and hexane) from the plant is prepared
and analyzed. Qualitative phytochemical tests are done to detect the presence of
Carbohydrate, Alkaloid, Tannins, Phenols, Saponins, Fixed oils, Gums and Mucilage.
Quantitative methods like phytochemical analysis, gravimetric estimation, RBC
membrane stabilization and TLC profiles are used to determine the active principle
Gymnemic acid, anti-inflammatory activity and antimicrobial activity. The results
showed that the leaf extracts studied contain the bio active compounds phenols,
alkaloids, tannins, saponins and Gymnemic acid. Water extract of normal var. had
positive reaction for carbohydrate (Molisch reagent) hairy var. leaves had negative
reaction. The presence of active phytochemical substances with antimicrobial and anti
-inflammatory activities may provide substantial basis for the use of this plant in
ethno medicine.

Journal of Research in Plant Sciences
An International Scientific Research Journal
INTRODUCTION:
India has been the rich source of plant based
drugs. Most of the Indian populations especially in rural
areas use traditional medicines only. On realizing the fact
that herbal medicine has no side effect (Bhandari and
Grover 1998, Shetty and Sunderasan, 1997) and many
more herbal medicines are reported to be effective
against many chronic disease for which there were no
allopathic drugs, production of herbal based medicine
has increased tremendously throughout the world. In
modern India extracts of Gymnema leaves are used
successfully, either alone or in combination with other
Ayurvedic compounds as a remedy against diabetes
mellitus. The term Destroyer of sugar is tradionally
used for Gymnema because chewing the leaves will
abolish the taste of sweetness. That is sweet food no
longer tasted sweet, but rather became almost completely
tasteless. The medicinally active parts of the plant are the
leaves and roots. Gymnema sylvestre is an Indian
medicinal plant used for diabetes in folk medicine,
Siddha and Ayurveda. Modern scientific studies also
proved its efficacy against the disease (Gent et al., 1999).
Gymnema sylvestre leaf extracts decreased the blood
sugar level in varying degrees. Blood sugar lowering unit
(BLU) of activity of each leaf extract
(Chatopadhyay;1999).Amrycard composed of Gymnema
sylvestre was tried in 20 patients of diabetes for three
weeks.Dose was determined on the basis of condition of
patients as 3 caps to insulin dependend case,2caps for
moderate case and 1 cap to mild diabetic case.It is
concluded that the drug provides a total beneficial
therepy in all type of diabetes and tends to increase
insulin secretion from islets of pancreas.(Bhagwan
singh;1997) Gymnema sylvestre is a liane.Two varities
exist in this species;one with normal and the other with
hairy leaf.Leaf in normal variety is lanceolate to ovate
apex acute and base truncate, less pubescent on both
sides.The hairy variant is ovate to cordate in shape, apex
acute and base cordate. With the above background the
present study investigates the pharmacological,
phytochemical properties and anti-inflammatory,
antimicrobial activities of the leaves of normal and hairy
variants of the medicinal plant Gymnema sylvestre.

MATERIALS AND METHODS:
Collection of plant material and identification:
The plant specimen Gymnema sylvestre were
collected from herbal garden, Tamil University,
Tanjavur. Collected plants were carefully examined and
identified with the help of Local Floras. Identities of
specimens are further confirmed with reference to
herbarium sheets at Rapinath Herbarium,
Trichy.Voucher specimen were deposited at Tamil
University, Herbarium.
Extraction:
The leaves of Variants Gymnema sylvestre are
shade dried and mechanically powdered after keeping it
in an oven at 40C for 24 hours. These powdered
materials are used for further physiochemical,
phytochemical and fluorescent analysis.
Preliminary phytochemical analysis:
Preliminary phytochemical analysis is done using
the procedures of Kokate (1994), (Balakumar etal; 1998).
Alkaloids, Carbohydrate, Tannins, Phenols, and
Saponins, Fixed oils, gums, mucilage and fats were
quantitatively analyzed. The intensity of the coloration
determines the abundance of the compound present.
Antimicrobial studies:
The aqueous extract of leaves Variants of
Gymnema sylvestre (Alagesaboopathi C. 2011) is used
throughout the study. The aqueous extract 10%, 20% and
40% are tested against different bacterial pathogens
(Microbial Type Culture Collection-MTCC) such as
Staphylococcus epidermis (MTCC-435), Klebsiella
pneumonia (MTCC-2653), Serratia marcens (MTCC-
2645), and Citrobacter diversus for its antimicrobial
activity. It was demonstrated by well diffusion assay.



Sangeetha and Jegadeesan, 2012
078 Journal of Research in Plant Sciences (2012) 1: 077-082
Physicochemical analysis:
Leaves of Gymnema sylvestre are analyzed for
ash value, loss on drying, extractive values, solubility
etc...In different solvents and extracts, using the
procedures recommended in Indian pharmacopoeia
(Anonymous, 1996; 1985; 1966). The colour of the leaf
powder of normal variety is dark green and that of hairy
variant is light green.
Fluorescent analysis: Fluorescent analyses of
leaves of Variants of Gymnema sylvestre in different
chemical reagents were analyzed using both Visible and
UV light.
TLC Profiles:
For TLC separation the alcoholic extracts of
leaves of Variants of selected medicinal plant Gymnema
sylvestre is spotted on silica gel-G coated plates.BAW,
Water, Ferosal and 60%Acetic acid was used as mobile
phase. Equal proportion of iodine chamber was used as
detecting reagent.

RESULTS AND DISCUSSION:
Physiochemical Analysis:
Physiochemical values of leaf of normal, hairy
var. and stored leaf are given in Table 5. It is interesting
to note that all the physiochemical values of normal leaf
and stored leaf differ greatly the values are also differ
between leaves of normal and hairy var. Total ash
content value of hairy var. is grater than that of normal
var. and among normal var. total ash content of fresh
leaves is less (13.7%) than stored leaves. Acid insoluble
ash value for normal var. is less (0.36%) than that for
hairy leaf (0.65%) in normal variant stored leaf
registered more acid insoluble ash value (0.41%). Water
soluble ash content of normal var. was far grater
(39.82%) than that of hairy var. (27.09%) and this value
in stored leaf was also lower than fresh leaves. Sulphated
ash value of normal var. was less (10.43%) when
compare to hairy var. (11.26%) and stored leaf (15.48%).
Normal var. had lower loss on drying. Solubility of leaf
of normal var. is more (29.96%) than that of hairy var.
(20.16%) and stored leaf (28.74%). Extractive value %
of normal var. is more (32.34%) compared to hairy and
stored leaf.
Qualitative phytochemical Analysis:
Leaf extract of Gymnema var. in water and
alcohol positively for all the phytochemical compounds
(Table 1). Though both water and alcohol extracts of
normal and hairy var. had similar phytochemical
Sangeetha and Jegadeesan, 2012
Journal of Research in Plant Sciences (2012) 1: 077-082 079
Table -1 Prelimnary phytochemical analysis of gymnema sylvestre
S. No.
Compound
Tested
Reagent
Water Alcohol
Normal
Normal
(stored)
Hairy Normal
Normal
(stored)
Hairy
1 Carbohydrate
Fehlings - - - - - -
Molish ++ ++ - + ++ ++
2 Alkaloid
Picric +++ +++ ++ ++ +++ +++
Mayer ++ ++ + + ++ ++
Wagners +++ +++ +++ +++ ++ -
3 Tannis & Phenols
Fecl
3
+++ +++ ++ ++ - -
Gelatin + + ++ ++ ++ ++
Lead acetate +++ +++ +++ +++ ++ -
4 Saponin Foam +++ +++ +++ ++ +++ +++
5 Gum & Mucilage - ++ ++ ++ ++ ++ ++
6 Fixed & Oils - + - + ++ ++ ++
(+++) - Appreciable Amount
(++) - Moderate
(+) - Small Amount
(-) - Completely Absent


constituents, some diagnostic characteristics were noted.
Water extract of normal var. had positive reaction for
carbohydrate (Molish reagent) hairy var. leaves had
negative reaction. Alcohol extracts of normal, stored and
hairy leaves answered positive for carbohydrates. It is to
be noted that water and alcohol extracts of the three leaf
samples had not answered for Fehlings reagent
indicating absences of reducing sugar. It was also
observed that stored leaf had not answered for oil and fat.
Anti Inflammatory:
The in-vitro anti-inflammatory study (RBC
membrane stabilization) using different concentrations
(10, 25, 50, 75, 100 & 200 g/ml) of Gymnema aqueous
extracts was carried out. It was found that leaf extract of
normal var. at a concentration of 200 g/ml showed
membrane stabilization activity with maximum activity
200 g/ml concentrations (Graph 1&2). It was found that
leaf extract of hairy var. also showed membrane
stabilization activity concentration 200 g/ml.
Antimicrobial activity:
Water extract of G. sylvestre normal and hairy
var. were screened for antibacterial and antifungal
activity against Staphylococcus epidermis, Klebsiella
pneumonia, Serratia marcens, Citrobacter diversus,
Aspergillus niger and Aspergillus flavus (Table 2).
Antimicrobial activity of aqueous extracts of both
variants revealed that all concentration inhibited growth
of S.epidermis with increase in zone of inhibitions with
increasing concentration. Leaf extract of normal
G.sylvestre had inhibitory activity against K.pneumonia
and A.niger whereas that of hairy var. had no activity
against these organisms. Both variant had activity against
S.marcens whereas hairy var. had inhibitory action
against C.diversus and A.flavus. Normal var. had no
activity.
TLC Studies:
Rf values of aqueous extracts of G. sylvestre leaf
(normal, hairy and stored) were determined on TLC
(Table 4). On TLC when 60% acetic acid was used as
mobile phase, all the three leaf samples had only one
spot Rf value 0.95. In BAW solvent phase, the normal
variant had two spots with Rf values 0.18 and 0.88.
Hairy var. had two spots with Rf values at 0.18 and 0.53.
Stored leaf had only spot corresponding with Rf value
0.16. TLC profile using Ferosal as mobile phase gave a
single spot with Rf value 0.74 for normal leaf, two spots
at Rf values 0.74 and 0.81 for hairy var. and two spots at
0.74 and 0.83 for stored leaf. In water, normal leaf, hairy
leaf and stored leaf exhibited only one spot with Rf value
0.79. Thus the three leaf samples of G.sylvestre had
distinguished.TLC profiles in the four mobile phase
used. Thus the three leaf samples had different profiles,
though they had common spots with similar Rf values.
Thus, as 60% acetic acid as solvent base, all the leaf
samples had only one spot with Rf value 0.95. In BAW,
the entire sample had a band at Rf 0.18. In Ferosal at Rf
0.74 all samples had spot. And in water, all the samples
had only one spot with Rf value at 0.79. This indicates
that while similar biological active compounds are
present in all the samples they differ by either presence
or absence of additional compounds.

Sangeetha and Jegadeesan, 2012
080 Journal of Research in Plant Sciences (2012) 1: 077-082
Table -2 Antimicrobial activity of aqueous extracts of gymnema sylvestre
S. No Organism
Zone of Inhibition (cm)
Concentration
10% 20% 30%
Normal Hairy Normal Hairy Normal Hairy
1 Staphylococcus epidermis 1.3 0.7 1.5 2.7 2.7 3.0
2 Klebsiella pneumonia 1.0 Nil 1.1 Nil 1.4 Nil
3 Serratia marcesens 1.0 0.9 1.1 1.9 2.1 1.5
4 Citrobactor diversus Nil 0.6 Nil 1.4 Nil 1.7
5 Aspergillus niger 1.4 Nil 2.0 0.8 2.2 1.1
6 Aspergillus flavus Nil 1.2 Nil 1.8 Nil 1.9
Leaves of normal and hairy variants of
Gymnema sylvestre varieties in water and alcohol had
answered positively for all the phytochemical
compounds (Table-1).
Anti-microbial activity (Table-2) of water
extracts of normal and hairy varieties of G.sylvestre was
screened. Both varieties had shown dosage dependent
inhibitory activity against S.epidermis .Normal varieties
had activity against K.pneumoniae,A.niger while hairy
variant had no activity. Hairy variant had action against
C.diversus and A.flavus. Thus two variants of
G.sylvestre have similar and dissimilar antimicrobial
activity against specific organisms.
Physicochemical, organoleptic, quantitative,
microscopical analysis of two varieties of G.sylvestre
revealed both similar and dissimilar characters which can
be used for diagnostic purpose.
Fluorescent analyses of leaf powder of normal
and hairy varities of G.sylvestre in different chemical
reagents. (Table-3) showed no major distinguishing
features for both Visible and UV light. This indicates
that both varieties have similar chemical constituents
Water extract of normal and hairy varieties
exhibited RBC membrane stabilizing activity (Siddiqui,
2000) at 200g/ml concentration. Thus both varieties
have anti-inflammatory activity (Graph-1&2).
Sangeetha and Jegadeesan, 2012
Journal of Research in Plant Sciences (2012) 1: 077-082 081
Table -3 Fluorescent analysis of dried leaf powder of gymnema sylvestre
S. No Powder + Reagent
Normal Var. Hairy Var.
Visible UV Visible UV
1 Powder Light Green Light Green Light Green Light Green
2 P + IN HCL Dark Green Dark Green Dark Green Dark Green
3 P + H
2
SO
4
Dark Brown Dark Brown Dark Brown Dark Brown
4 P + HNO
3
Yellow Light Yellow Yellow Dark Yellow
5 P + NH
4
OH Dark Green Dark Green Dark Green Dark Green
6 P+Picric Acid Light Green Light Green Light Green Light Green
7 P+Iodine Light Brown Dark Brown Brown Brown
8 P+FeCl
3
Black Brown Black Brown
Table -4 R values of aqueous extract of gymnema sylvestre leaf
Mobile Phase
R Value of Gymnema Sylvestre
Normal Hairy Normal (Stored)
Spot-I Spot-II Spot-I Spot-II Spot-I Spot-II
Acetic Acid 0.95 - 0.95 - 0.95 -
BAW 0.18 0.88 0.18 0.53 0.18 -
Ferosal 0.74 - 0.74 0.81 0.74 0.83
Water 0.79 - 0.79 - 0.79 -
Graph 1: Effect of aqueous extract of Gymnema
Sylvestre (Normal) on RBC Lysis
O
D

A
T

5
6
0

m
m

Graph 2: Effect of aqueous extract of Gymnema
Sylvestre (Hairy) on RBC lysis
O
D

A
T

5
6
0

m
m

Sangeetha and Jegadeesan, 2012
082 Journal of Research in Plant Sciences (2012) 1: 077-082
TLC profiles of aqueous extracts (Table-4) of
normal, hairy and stored leaves samples of G. sylvestre
revealed diagnostic features. All the three leaf samples
developed only one spot in acetic acid and water,
whereas in BAW and ferosol three samples developed
different bands with different R values. The TLC study
is a further proof that these two variants are
phytochemically different.

REFERENCES
Alagesaboopathi C. 2011. Antimicrobial screening of
selected medicinal plants Tamilnadu, India. Afr J
Microbiol Res., 56:617-621.

Balakumar T and selvakumar. 1998. Prelimnary
observation on the micro propagation of the antidiabetic
herb Gymnema sylvestre.National Plant Research.A
(52):2-4.

Bhandari V and Grover JK. 1998. A reappraised of
clinical studies on the comparative influence of three
indigenous plant drug in diabetes melliyus.Hamard
me d i c u s . 4 1 ( 2) ; 9 - 1 5. En vi bbs i , 20 0 4. h t t p: / /
www.envibbsi.org/medi:htm#Gymnema sylvestre.

Debruyne D. 1997. Clinical pharmacokinetics of
fluconazole in superficial and systemic mycoses. Clin
Pharmacokinet., 33:52-77.

Gent JF, Persand,Hettinger TP, Frank ME and
marks CE. 1999. Tast confusion following gymnemic
acid rinse,chem. Senses.24:393-403.

Shetty BR and Sunderasan. 1997. Studies on the
resayana effect of a geriatric formulation in apparently
normal aged persons.Journal of Research Ayurveda and
Siddha.18 (3-4):108-117.

Siddiqui. 2000. Development in the chemistry and
pharmacology of G.sylvestre.Journal of Medicinal and
aromatic Plant sciences.22. (2-4):222-231.


S.No Parameter Normal
Normal
(Stored)
Hairy
1. Total Ash 13.71 15.50 17.88
2.
Acid-insoluble
Ash
0.36 0.41 0.65
3.
Water-soluble
Ash
39.82 18.00 27.09
4. Sulphate Ash 10.43 15.48 11.26
5.
Loss on
Drying
3.23 - 4.44
6.
Solubility
Water 29.96 28.74 20.16
Alcohol 10.26 36.02 22.52
7.
Extractive
Value Water
32.34 26.62 22.89
8.
Gymnemic
Acid
2.75 3.7 3.75
Table-5 Physiochemical Values (Precentage) of
Leaf Powder Gymnema Sylvestre
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