International Journal of Medicine and

Pharmaceutical Science (IJMPS)

ISSN(P): 2250-0049; ISSN(E): 2321-0095
Vol. 5, Issue 6, Dec 2015, 59-64
TJPRC Pvt. Ltd.

EXTRACTION OF SECONDARY METABOLITES AND ANTIMICROBIAL

STUDIES ON MOMORDICA.CYMBALARIA (FENZYL EX NAUDIN)
AN ENDANGERED MEDICINAL PLANT
PAVITHRA. S1, K. PAVANA2 & DEVENDRA. B. N3
1,2,3
1

Department of Biotechnology, Navodaya College of Biosciences, Raichur, Karnataka, India

Department of Biotechnology, Little Flower Degree College, Uppal Cross Roads, Hyderabad, Telangana, India

ABSTRACT
Momordica. Cymbalaria is a medicinal plant of family Cucurbitaceae, and is herbaceous annual or perennial
climber, commonly used as medicinal vegetable for cooking purpose. Generally it is distributed in tropical zones of
Deccan plateau. In Karnataka, it is found in Raichur, Bellary, Bijapur, and Gulbarga regions, well known as
karchikayee. The present study was carried out to explore the phytochemical constituents and antimicrobial activity of
fruit Momordica. Cymbalaria in comparison with standard antibiotics. The antimicrobial activity of petroleum ether,
chloroform, ethanol and aqueous extracts from fruit of the plant was studied using Escherichia. Coli, Salmonella typhi,

all the five microorganisms. The result reveal that the fruit of M. cymbalaria has very good anti-microbial activity
against gram negative and gram positive organism when compared to standard antibiotics. The chloroform and ethanol
extract has shown significant activity against all the organisms but rather more towards Escherichia Coli, Salmonella
typhi. Similarly, petroleum ether and aqueous extract of plant also showed good inhibition against Vibrio cholera and

Original Article

Staphylococcus aureus, Vibrio cholera, Klebsiella pneumonia as test organisms, all the extracts were effective against

klebsiella. This indicates the fruit of M. cymbalaria has potential antimicrobial activity with concentration dependent
and can fight against these organisms and could be a good substitute to the contemporary medicines.
KEYWORDS: Momordica cymbalaria, Antimicrobial Activity, Phytochemical Constituents

Received: Nov 21, 2015; Accepted: Dec 08, 2015; Published: Dec 11, 2015; Paper Id.: IJMPSDEC20158

INTRODUCTION
Medicinal plants are the most important natural source of life saving drugs for the majority of the worlds
population. They have been the subject of mans curiosity since time immemorial. Almost all civilization has a
history of medicinal plants and their applications. The search for components with antimicrobial activity has
gained increasing importance in recent times due to growing worldwide concern about the alarming increase in the
rate of infections caused by multi drug antibiotic resistant strains of microorganisms, which adds urgency to search
for new infection fighting strategies. Plants provide a valuable material base for the discovery and development of
new drugs with natural origin without many side effects, besides having enormous therapeutic potential of healing
many infections and diseases (Marjorie MC et al., 1999). This therapeutic property of plants lies in chemical
substance known as secondary metabolites, which include a wide variety of phytochemicals.
Momordica Cymbalaria is a medicinal plant and its taxonomy is as follows;

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Pavithra. S, K. Pavana & Devendra. B. N

Kingdom: Plantae
Subkingdom: Tracheobionata
Super Division: Spermatophyta
Division: Magnoliophyta
Class: Magnoliopsida
Subclass: Dilleniidae
Order: Cucurbitales
Family: Cucurbitacea
It is herbaceous annual or perennial climber, generally distributed in tropical zones of Deccan plateau. In
Karnataka, it is found in Raichur, Bellary, Bijapur, and Gulbarga regions and is well known as karchikayee, commonly
found in fences as shrubs and in hedgerows (R C Grewal., 2003). It is a major family for economical important species
particularly those with edible fruits, and is distinct from other families both morphogically and biochemically. Therefore, it
is considered as monophyletic. It is trailing monoecious herb often tuberous, tendrils simple, leaf blade reniform, orbicular
3-5 angled or lobed, sparely hairy, cordate at the base, staminate flower 4-5, stamens 2, fruit fusiform, shortly beaked,
pubescent, longitudinally ribbed 1.8-4 cm long, flowering occurs from June to October, fruits are used as vegetables.
Momordica Cymbalaria Fenzyl ex Naudin contains many medicinal values from folklore; its roots are used as an
abortifacient (chorpa et al., 1956). The green fruit was also used to increase body heat, salivation and it is also evident of
having properties like anti-helminthics, anti-diarrhoeic and to cure burning soles (Rao, Kesavulu, et al., (2003). R. Jeyadevi
et al., (2012) reported that Momordica Cymbalaria has Antidiabetic and hypoglycaemic activity, Hepatoprotective activity,
Cardioprotective effect, Anti diarrhoeal activity, Anti ulcer activity, Nephroprotective activity, Anti cancer activity, Anti
implantation and anti ovulatory activity.
The versatile medicinal activity of this plant urged us to extract secondary metabolites from the fruit of
Momordica Cymbalaria and determine its antimicrobial activity against few gram negative microorganisms such as,
Escherichia Coli, klebsiella pneumonia, Vibrio cholera, Salmonella typhi and gram positive (Staphylococcus aureus)
microorganisms.

MATERIALS AND METHODS

Plant Material
The plant material Momordica Cymbalaria (Fenzyl ex Naudin) (Figure 1) used for this study was collected from
fields of Manvi Taluk, Raichur District, Karnataka, India. The plant specimen was authentically identified with the help of
flora of Gulbarga district (Seetharama et al., 1998).Figure 1.

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Extraction of Secondary Metabolites and Antimicrobial Studies on Momordica

cymbalaria (Fenzyl Ex Naudin) an Endangered Medicinal Plant

61

Figure 1: Photograph of Momordica cymbalaria A) Plant and B) Fruits

Extraction of Secondary Metabolites from Momordica cymbalaria
The Fruit of Momordica Cymbalaria was shade dried and grinded to a fine powder and further used for extraction
of secondary metabolites using various solvents such as petroleum ether, chloroform, ethanol (95 %) and distilled water
successively through cold extraction.
Phytochemical of Screening of Momordica cymbalaria
The fruit of Momordica Cymbalaria was air dried and were qualitatively tested for presence of various
photochemical using various standard tests such as test for phenols (Gibbs 1974), test for flavanoids (Peach and Tracey
1959), test for steroids (Gibbs 1974), test for tritepenoids, test for tannins (Trease and Evens 1980), test for glycosides
(Kokate et al., 1997), test for Saponins (Gibbs 1974), test for Alkaloids (Meyers test, Wagners test, Dragendroffs test).
These tests were based on visual observation of colour modification and precipitation upon the addition of specific
reagents. (Table 1)
Separation of Secondary Metabolites of Momordica cymbalaria by Thin Layer Chromatography (TLC)
Secondary metabolites extracted from fruits of Momordica Cymbalaria were subjected to TLC according to the
technique formulated by Ergon Stahl (1965). In this technique glass plate of size 20 x 20 cm was used to prepare
chromatographic plates using silica gel G, plant extract was loaded on to the TLC plates with the help of capillary tubes.
Development of Chromatographic Plates
The chromatographic plates were separated using different solvent system such as Ethyl acetate: Ethanol and
Water, with the ratio (3:2:5), Chloroform: Butanol: Water, with the ratio (4:3:1), and Chloroform: Water, with the ratio
(7:3). The plates were loaded with sample containing secondary metabolites of Momordica Cymbalaria, air dried and these
plates was placed vertically in the solvent, once the solvent reaches about 18 cm, the plates were removed, air dried and
then the retention factor (Rf-value) of the spots was calculated.
Antimicrobial Studies on Momordica cymbalaria
The successive extracts such as petroleum ether extract, chloroform extract, ethanoloic extract and aqueous
extract of Momordica cymbalaria, was concentrated by distilling the solvent through evaporation, air dried and then
dissolved in Dimethyl sulfoxide (DMSO) at a concentration of 1mg/ml. Antimicrobial screening was done using agar
diffusion method. Nutrient agar plates were prepared and the bacterial samples of 0.1ml were spread uniformly on to the
plates, wells were made with help of crock borer and 0.1ml of crude drug extract dissolved in DMSO was loaded as
bioassay for studying the antimicrobial activity of Momordica cymbalaria against few gram negative and gram positive
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Pavithra. S, K. Pavana & Devendra. B. N

pathogenic microorganisms such as Escherichia Coli, klebsiella pneumonia, Vibrio cholera, Salmonella typhi and
Staphylococcus aureus obtained from samples collected from Department of Microbiology, Navodaya Medical College and
Hospital, Raichur, Karnataka, India, and the results were recorded in terms of diameter of zone of inhibition(mm). The
above results were compared using standard antibiotic streptomycin at a concentration 1mg/ml as a control.

RESULTS AND DISCUSSIONS

Preliminary Screening of Secondary Metabolites and Antimicrobial Activity of Momordica cymbalaria
In the present study an effort was made to screen the presence of secondary metabolites from the fruits of
Momordica Cymbalaria. The results of preliminary screening of secondary metabolites are given in the table 1, and the
range of antimicrobial activity are represented graphically in Figure 2, this clearly depicts that all the extracts of
Momordica Cymbalaria shows antimicrobial activity against all the test organisms, The chloroform and ethanol extract has
shown significant activity against all the organisms but rather more towards Escherichia Coli (14and 9 mm) Salmonella
typhi (11 and 14 mm), Staphylococcus aureus (8 and 7 mm), Vibrio cholera (10 and 5 mm), klebsiella pneumonia (8 and 6
mm). Similarly, petroleum ether and aqueous extract of plant also showed good inhibition against Vibrio cholera (12 and 3
mm) and klebsiella pneumonia (7 and 5 mm), Salmonella typhi (7 and 4 mm), Staphylococcus aureus (6 and 4 mm) against
standard antibiotic streptomycin inhibition ranging from 7-19 mm. This is due to the presence of secondary metabolites
like steroids, terpentoids, tannins, resins and alkaloids in chloroform extract. The antimicrobial activity of ethanolic extract
is due to the presence of secondary metabolites like phenols, flavonoids, terpenoids, and glycosides.
Table 1: Preliminary Screening of Secondary Metabolites
S No

Tests

P.E
Extract

C.E
Extract

E.E
Extract

A.E
Extract

Test for Phenols

a.
Phenol test
+
b.
Ellagic acid test
+
Test for Flavonoids
2
a.
Flavonoid test
+
b.
Lead acetate test
+
Test for Steroids
3
a.
Acetic anhydride
+
b.
Chloroform
+
Test for Tripenoids
a.
Salkowiski test
4
+
+
b.
Libermann
+
+
Burchards test
Test for Tannins
5
a.
Ferric chloride test
+
b.
Gelatin test
+
Test for Glycosides
6
Kellar killanis test
+
7
Resins
+
+
Saponins
8
Foam test
+
Test for Alkaloids
a.
Mayers test
+
9
b.
Dragendroffs test
+
c.
Wagner test
+
P.E= Petroleum, ether; C.E= chloroform, ethanol; E.E= Ethanol extract; A.E= Aqueous extract; -= negative;
1

+= Positive;
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Extraction of Secondary Metabolites and Antimicrobial Studies on Momordica

cymbalaria (Fenzyl Ex Naudin) an Endangered Medicinal Plant

63

Figure 2: Anti-Microbial Activity of Various Extracts of Momordica cymbalaria on different

Microorganisms and their Comparative Study with Antibiotic Streptomycin as Control
P.E= Petroleum, ether; C.E= chloroform, ethanol; E.E= Ethanol extract; A.E= Aqueous extract.
E.Coli: Escherichia Coli, S.typhi: Salmonella typhi, S. aureus: Staphylococcus aureus, V.cholera: Vibrio cholera,
Klebsiella: klebsiella pneumonia
Separation of Secondary Metabolite by TLC
Among various chromatographic techniques thin layer chromatography (TLC) is extensively employed for rapid
drug analysis and subsequent drug preparation and very much useful in monitoring the identity and purity of drugs and also
provides semi quantitative information on major active constituents of a drug. TLC was performed on successive extracts
of petroleum ether, chloroform, ethanol, and aqueous extract of Momordica cymbalaria. In which petroleum extract
displayed one distinct band possessing light green colour with Rf value 0.45, where as the chloroform extract displayed
two distinct bands possessing orange colour with Rf value 0.5 and green colour band with Rf value 0.68 and the ethanolic
extract displayed one distinct band possessing orange green colour with Rf value 0.8.

CONCLUSIONS
Worldwide increase of multidrug resistant in both community and health care associated infections has justified
the search for alternatives, for considering plants as an potentially important useful material for the development of natural
therapeutic agents is the need of the hour, hence in this regard we intended to find out the invitro antimicrobial activity of
Momordica cymbalaria which has confirmed to inhibit the growth of few gram positive and gram negative test organisms,
in which all the extracts of Momordica cymbalaria showed antimicrobial activity from which the chloroform and ethanolic
extract showed major significance. Momordica cymbalaria is disappearing from the habitat at an alarming rate as now it
has been one of the endangered plant species in India. This clearly opens up a huge challenge to conserve and manage our
natural resource wisely. Based on the results obtained it is evident that the medicinal plant Momordica cymbalaria has
antimicrobial activity against various microorganisms used in this study.

ACKNOWLEDGEMENTS
The authors are thankful to Dr.T.Srinivas, Registrar, Navodaya College of Medical Sciences, Raichur, Karnataka,
India, for his support and encouragement and the management for providing the facilities for execution of this research
work.

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Pavithra. S, K. Pavana & Devendra. B. N

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