INVITRO ANTIBACTERIAL ACTIVITY OF METHANOLIC ... - IJPRD

Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-5/JULY/007 ISSN 0974 – 9446
INVITRO ANTIBACTERIAL ACTIVITY OF METHANOLIC ROOT EXTRACT OF
TINOSPORA CORDIFOLIA (WILLD)
ABSTRACT
M. Fatima Rose 1 *, K.M.Noorulla 2 ,
M.Asma 4 ,R.Kalaichelvi 5 ,
K. Vadivel 1 ,B.. Thangabalan 1 , B.N.Sinha 5
1
M. Fatima Rose
SIMS College of Pharmacy, Mangaldas Nagar,
Guntur-522 001, India
4
C.L.Baid Metha College of Pharmacy, Thorapakkam, Chennai-600096, India
3
Ayya Nadar Janaki Ammal College, Sivakasi -626123, India
4
K.C.Reddy Institute of Pharmaceutical Sciences, Jangamguntla Palem,
Medikonduru Mandal, Guntur-522 348, India
5
Birla Institute of Technology, Mesra, Ranchi -835215, India
Email: rosepharma@gmail.com
The antibacterial activities of the hot and cold methanol extracts of the roots of Tinospora cordifolia
(Willd) Miers was evaluated on bacterial strains like Staphylococcus aureus, Shigella dysenteriae,
Escherichia coli and Pseudomonas aeruginosa. The in vitro antibacterial activity of hot and cold
methanol extracts was performed by cup plate agar diffusion method using ciprofloxacin (ciprozol-500)
in dimethyl sulphoxide as a standard drug for the comparision of antibacterial activity. From the
experiment done the hot methanol extract of Tinospora cordifolia (Willd) Miers did produce
considerable antibacterial activity than the cold maceration extract was observed. The maximum
antibacterial activity of hot and cold methanol extracts was exhibited against Staphylococcus aureus
when compared with standard drug. In addition the preliminary phytochemical tests of the hot and cold
methanol extracts of Tinospora cordifolia roots revealed the presence of alkaloids, carbohydrates,
flavonoids, glycosides, lignin, saponins, terpenes, tannins, steroids and reducing sugar. The results
obtained suggest that Tinospora cordifolia roots can be used in treating diseases caused by the test
organisms.
Key Words: Tinospora cordifolia (Willd), hot and cold methanolic extracts, ciprofloxacin, antibacterial
activity.
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Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-5/JULY/007 ISSN 0974 – 9446
INTRODUCTION
Tinospora cordifolia (Willd) Miers
(Menispermaceae) is a common climbing shrub,
found throughout India in forests. It is one of the
most valuable Traditional Indian medicinal
herbs and has been used in Ayurvedic
preparations for the treatment of various
ailments throughout the centuries [1]. Tinospora
cordifolia is reported to possess antispasmodic,
anti-inflammatory, antiallergic and anti-diabetic
properties. It is generally prescribed in general
debility, diabetes, fever, jaundice, skin diseases,
rheumatism, urinary diseases, dyspepsia, gout,
gonorrhoea and leucorrhoea [2]. A decoction of
the stems, leaves and roots is used to treat
fever, cholera, diabetes, rheumatism and
snake-bites, an infusion of the stem is drunk as
a vermifuge, A decoction of the stem is used for
washing sore eyes and syphilitic sores. The
stem is registered in the Thailand
Pharmacopoeia, and commonly use in hospital
to treat diabetes [3]. The stem decoction is
considered antipyretic, useful as an antimalarial
and a wash for skin ulcers. Traditionally
an infusion is used to treat fever due to malaria
and also in cases of jaundice and for use
against intestinal worms [4]. A decoction of the
fresh root mixed with pepper and goat’s milk is
given for rheumatism, where the dose is half a
pint every morning. The leaves are given for the
cure of gonorrhoea and is said to soothe the
smarting and scalding. It is also used externally
as a cooling and soothing application in prurigo,
eczema, impetigo [5]. The methanolic stem
extract of Tinospora cordifolia possesses
antifertility activity, which might be exploited to
prevent unwanted pregnancy and control the
ever increasing population explosion [6].
Decoction of the root in combination with ginger
and sugar is given in cases of bilious dyspepsia
and in cases of fevers with other bitters and
aromatics. Roots rubbed with bonduc nuts in
water are given for stomachache, especially in
children. They can also be used to treat
stomachache and jaundice. The infusion is also
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useful in fevers caused by smallpox and
cholera. Another popular use of this infusion is
in a mixture for treating indigestion. The leaves
are beaten with honey and applied to ulcers.
Stem, root, whole plant is used in the treatment
of wound, anthrax, pneumonia, asthma, and
cough [7]. The root of this plant is known for its
anti-stress, anti-leprotic and anti-malarial
activities [8]. The stem of Tinospora cordifolia is
one of the constituents of several Ayurvedic
preparations used in general debility,
dyspepsia, fever and urinary diseases [9, 10]. In
this article, we wish to report the antibacterial
activities of the methanolic root extract of
Tinospora cordifolia.
MATERIALS AND METHODS
Collection and Authentication of the plant
material :
The roots of Tinospora cordifolia (Willd) Miers
were collected in the month of August- 2007
from the campus of Birla Institute of
Technology, Mesra, Ranchi. The plant was
authenticated by Birsa Agricultural University,
Kanke, Ranchi; ref.no.686/Hort.,Kanke.
Preparation of extract
Hot continuous methanol extraction:
The shade dried coarse powder of the root
(750gm) was packed well in soxhlet apparatus
and was subjected with Methanol by continuous
hot extraction for about 48hrs. The extract was
filtered through whatmann filter paper and
concentrated on a water bath. Obtained extract
(Dark Reddish Brown) was weighed and % yield
was found to be 19.41%.
Cold methanol extraction:
The shade dried coarse powder of the root
(750gm) was kept in stoppered flask and were
macerated with Methanol for 24-48 hrs with
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Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-5/JULY/007 ISSN 0974 – 9446
frequent stirring. Then the extract was filtered
through whatmann filter paper and concentrated
under air. Obtained extract (dark reddish brown)
was weighed and % yield was calculated as
11.33%.
The above prepared hot and cold methanol
extract was stored in desiccators for further
phytochemical and antibacterial investigations.
The dried extract was tested for its
phytoconstituents by standard methods [11&12]
and the results were tabulated in Table 1.
The both methanolic extracts were diluted with
DMSO to the concentration of 10mg/ml,
20mg/ml, 30mg/ml, 40mg/ml and 50mg/ml.
Microorganisms like Staphylococcus aureus,
Shigella dysenteriae, Escherichia coli and
Pseudomonas aeruginosa were used for
testing.
Antibacterial evaluation
Cup plate agar diffusion method was used for
the antibacterial study. Working stock was
prepared as 1ml of each bacterial strain was
initially inoculated in100 ml of sterile nutrient
broth and incubated for 37°±1°C for 24 hr
respectively. The 0.2ml of the each test
organisms from the working stock were seeded
into 100ml sterile nutrient agar medium and
cooled to 48°C to 50° C in a sterile Petridish
respectively. When the nutrient agar medium
solidifies, four holes of uniform diameter (7 mm)
were made using sterile aluminium borer. Then,
0.2 ml of each hot, cold methanol extracts,
standard solution (10, 20,30,40,50 mg/ml)
respectively and vehicle control (DMSO) were
placed in each hole separately under aseptic
condition. The plates were then maintained at
room temperature for 2 hr to allow the diffusion
of the solution into the medium. All the bacterial
plates were then incubated at 37°±1°C for 18 hr
and the zone of inhibition was measured [cm,
including the diameter of the bore (7 mm)] and
the results were given in Table 2 and fig 1.
RESULTS AND DISCUSSION
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Results of the antibacterial screening of hot and
cold methanol extracts of Tinospora cordifolia
root revealed significant antibacterial activity
against all tested bacterial strains. The
maximum antibacterial activity of hot and cold
methanol extracts was exhibited against
Staphylococcus aureus when compared with
standard drug. The hot methanol extract of
T.cordifolia root showed more significant activity
against all tested bacterial organisms than that
of the cold methanol extracts. Preliminary
phytochemical analysis of methanolic extract
revealed the presence of alkaloids,
carbohydrates, reducing sugar, glycosides,
steroids, terpenes, lignin, saponins, tannins and
flavonoids. The inhibitory effects of this
medicinal plant on the test microorganisms may
therefore, be due to the presence of the above
phytochemical components. The isolation of
components of root of Tinospora cordifolia is in
progress as potent antibacterial agents.
CONCLUSION
The results obtained in this study suggests that
the hot methanol extract of T.cordifolia root
showed more significant activity against all
tested bacterial organisms than that of the cold
methanol extracts. These promissory extracts
open the possibility of finding new clinically
effective antibacterial compounds.
ACKNOWLEDGEMENTS
The authors are very much thankful to Dr. S.A.
Azeez, Principal, SIMS College of Pharmacy,
for his encouragement and support.
.
REFERENCES:
1. Indian medicinal plants-A Compendium of
500 species, Vol 1,Orient Longman Pvt Ltd,
Arya vaidya sala, Kottakkal.
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Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-5/JULY/007 ISSN 0974 – 9446
2. Kirtikar KP, Basu BD, Indian Medicinal
Plants, 2 nd ed, Vol 1, New Connaught Place,
Dehra Dun, 1975.
3. Nadkarni KM, Nadkarni AK, editors. Indian
Materia Medica, 3rd ed, Vol 1, Mumbai, M/S
Popular Prakasan Pvt. Ltd; 1976.
4. Khory RN, Katrak NN, Prakash K, Materia
Medica of India and their therapeutics, 2nd ed.
Delhi, 1984, 31.
5. Anonymous. Wealth of India: Raw materials,
Vol X. New Delhi: CSIR; 1976.
6. Gupta RS, Sharma A. Antifertility effect of
Tinospora cordifolia (willd) Stem
extract in male rats. Indian J Exp Biol. 2003, 41:
885-89.
7. Singh SS, Pandey SC, Srivastava S, Gupta
VS, Patro B, Ghosh AC, Chemistry And
Tables and Figures:
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Medicinal Properties of Tinospora cordifolia
(Guduchi). Indian Journal of Pharmacology
2003, 35: 83-91.
8. A Brief review of Ayurvedic concepts
Tinospora htm.
9. Aiyer KN, Kolammal M, editors.
Pharmacognosy of Ayurvedic drugs, Series1.1st
ed. Trivendram: The Central Research Institute;
1963.
10. Khosa RL, Prasad S. Pharmacognostical
studies on Guduchi (Tinospora cordifolia Mires).
J Res Ind Med 1971, 6: 261-269.
11. Trease GE, Evans WC, Pharmacognosy,
ELBS publication, New Delhi, 1985, 217.
12. Vaishali T, Makde KH, Padhye MD,
J.Nat.Prod. 2000, 16: 14-17.
Table 1: Phytochemical screening of methanolic root extract of Tinospora cordifolia.
S.No Phytoconstituent Hot Methanol Cold Methanol
Extract
Extract
1 Alkaloids +++ ++
2 Carbohydrates ++ +
3 Reducing Sugar ++ +
4 Glycosides + +
5 Steroids +++ ++
6 Terpenes +++ +
7 Lignin + +
8 Saponins ++ +
9 Tannins + +
10 Flavonoids + +
+++ Prominently Present, ++ Moderately Present, + Slightly Present.
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Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-5/JULY/007 ISSN 0974 – 9446
Table 2: Evaluation of antibacterial activity of the methanolic root extract of Tinospora cordifolia
Diameter of the zone of inhibition in (cm)
Extracts
tested
Conc.
(mg/ml) S.aureu
s
S.dyssenteri
ae
E.coli
(G-)
P.aurigenosa
(G-)
(G+) (G-)
10 2.0 2.0
1.7 2.1
20 2.3 2.2
2.1 2.1
Hot
30 3.0 2.7
2.2 2.2
methanol 40 3.3 2.8
2.3 2.5
extract 50 3.7 3.0
2.6 2.7
10 2.0 1.9
1.5 1.7
20 2.2 2.1
1.8 1.9
Cold 30 2.4 2.2
1.9 2.1
methanol 40 2.5 2.3
2.2 2.4
Extract 50 2.9 2.5
2.3 2.5
10 3.7 3.2
3.1 2.8
Ciprofloxacin 20 4.0 3.5
3.2 3.1
30 3.9 3.0
3.4 3.2
40 4.1 3.1
3.5 3.4
50 4.2 3.3
3.6 3.7
(G+) gram positive bacteria, (G-) gram negative bacteria
Fig 1. Antibacterial activity of the hot and cold methanol extracts of Tinospora cordifolia root.
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