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M.Thenmozhi., et al. / International Journal of Advances in Pharmaceutical Research
IJAPR
Available Online through
www.ijapronline.org
Research Paper
ISSN: 2230 – 7583
PHYTOCHEMICAL AND ANTIMICROBIAL EVALUATION OF ECLIPTA ALBA
M.Thenmozhi* and Rajeshwari Sivaraj
Department of Biotechnology, Karpagam University, Coimbatore-641021, Tamilnadu, India,
thenmozhibio@yahoo.co.in and rajeshwarishivaraj@gmail.com
Received on 22 – 04 - 2011 Revised on 25 – 05- 2011 Accepted on 05 – 06 – 2011
ABSTRACT
The increasing prevalence of multidrug resistant strains of bacteria and the recent appearance of strains
with reduced susceptibility to antibiotics raises the urgency to the search for new infection-fighting strategies. The
World Health Organisation (WHO) estimated that 80% of the population of developing countries still rely on
traditional medicines, mostly plant drugs, for their primary health care needs. Medicinal plants thus continue to be
an important therapeutic aid for alleviating ailments of human kind. In the present study, plant from Asteraceae
family named Eclipta alba was taken and analysed phytochmical test. The plant was extracted with different
solvents and the antimicrobial activity checked using ten bacterial and fungal cultures using agar well diffusion
method. The assessment of antimicrobial activity was based on the measurement of the concentration diameter of
the inhibition zone formed around the well. The methanolic extract was having the best activity.
Key Words: phytochmical analysis, antimicrobial activity, Asteraceae, Eclipta alba
INTRODUCTION
In the modern medicinal system, people
have started looking at the ancient healing systems
like Ayurveda, Siddha and Unnani due to the adverse
effects associated with synthetic drugs. Herbal drugs
play an important role in health care programs
especially in developing countries. Ancient Indian
literature incorporates a remarkably broad definition
of medicinal plants and considers ‘all ‘plant parts to
be potential sources of medicinal substances 1 . Plants
have the capacity to synthesise a diverse array of
chemicals. The understanding of how phytochemicals
function in plants may further increase our
understanding of the mechanisms by which they
benefit humans.
For Correspondence:
M.Thenmozhi
Department of Biotechnology,
Karpagam University,
Coimbatore-641021,
Tamilnadu, India.
e.mail: thenmozhibio@yahoo.co.in
Ph: 09788851777
In plants, these compounds attract beneficial and
repel harmful organisms as photoprotectants and
respond to environmental changes 2 . A main stream
medicine is increasing the use of antimicrobials and
other drugs derived from plants as traditional
antibiotic 3 . Screening active compounds from plants
has lead to the discovery of new medicinal drugs
which have efficient protection and treatment roles
against various diseases, including cancer and
Alzheimer’s disease 4 . Infectious diseases are the
leading cause of death world wide and antibiotic
resistance has become a global concern 5 .
There has been an increasing incidence of
multiple resistances in human pathogenic micro
organisms in recent years, largely due to
indiscriminate use of commercial antimicrobial drugs
commonly employed in the treatment of infectious
diseases. This has forced scientists to search for new
antimicrobial substances from various sources like
the medicinal plants (IWU, 2002). A few antifungal
agents are available and licensed for use in veterinary
practice or human treatment. The use of systemic
drugs is limited to treat man or animal due to their
high toxicity and problems of residues in products
intended for human consumption 6 . The plant used for
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M.Thenmozhi., et al. / International Journal of Advances in Pharmaceutical Research
the present study was Eclipta alba, used for memory
disorder treatment, edema, fevers and rheumatic joint
pains treatment, digestion, hepatitis, enlarged spleen,
antioxidant activity and skin disorders and as general
tonic 7 .
Over the past twenty years, there has been a
lot of interest in the investigation of natural materials
as sources of new antibacterial agents 8, 9 . Different
extracts from traditional medicinal plants have been
tested and many reports have shown the effectiveness
of traditional herbs against microorganisms 10 . The
increasing interest on traditional ethno medicine may
lead to discovery of novel therapeutic agents.
Medicinal plants are finding their way into
pharmaceuticals, neutralceuticals, cosmetics and food
supplements. In this regard, plants have given
western pharmacopoeia of about 7000 different
pharmaceutically important compounds and a number
of top-selling drugs of modern time, e.g. quinine,
artemisinin, taxol, camptothecin, etc., 11 . Until natural
products have been approved as new antibacterial
drugs, there is an urgent need to identify novel
substances active towards highly resistant
pathogens 12,13 . Therefore, the present study is
designed to evaluate the antimicrobial activity of
different extracts of the leaves of Eclipta alba.
MATERIALS AND METHODS
Collection of plant materials: The plants of Eclipta
alba were collected from different places of
Namakkal District, Tamilnadu. The plants were
identified at Botanical Survey of India, South circle,
Coimbatore.
Preparation of Plant samples: The plants collected
were washed thoroughly under running tap water and
then were rinsed with distilled water. Then plants
were then shade dried without any contamination for
about 3 to 4 weeks. The dried plant sample was
powdered in blender and stored in airtight containers.
Phytochemical screening
Phytochemical screening was carried out to
assess the qualitative chemical composition of crude
extracts using commonly employed precipitation and
coloration to identify the major natural chemical
groups such as carbohydrates, cardio glycosides,
saponins, oils and fats, terprnoids, alkaloids, steroids
and sterol, flavonoids, tannins and phenolic
compounds, amino acids and proteins, quinones.
General reactions of the analysis revealed the
presence or absence of these compounds in the crude
extracts tested. Crude extracts of the plants were
previously prepared and stored in a refrigerator for
phytochemical tests 14 .
Preparation of Plant extracts:
About 50 g of dried powdered samples was
soaked in 125 ml of petroleum ether for 16 hours in a
rotatory shaker. Whatman No.1 filter paper was used
to separate the extract of both plants. The residue was
again dried and was soaked again in chloroform for
16 hours and extracted, the same was continued with
the next three solvents namely ethyl acetate,
methanol and water (aqueous). These filtrates were
used for further analysis and tests 15 .
Antimicrobial activity
Microorganisms and Culture media used for
study
Ten bacterial and fungal cultures were
bought from Department of Microbiology, Karpagam
University, Coimbatore and was mother cultured and
maintained in the Department of Biotechnology
(Karpagam University, Coimbatore). Nutrient agar
was used for the mother culture preparation of
bacterial cultures. The broth for bacteria was
maintained in Nutrient broth and plates were
prepared in Nutrient agar. Potato dextrose agar was
used for maintaining the mother culture and pure
culture of fungal species. Plates were also prepared in
Potato dextrose agar. Ten bacterial cultures namely
Bacillus cereus, Salmonella typhi, Proteus vulgaris,
Klebsiella pneumoniae, Citrobacter freundii,
Escherichia coli, SERRATIA MARCESCENS,
Streptococcus pyogenes, Staphylococcus aureus and
Pseudomonas auregenosa and ten fungal culture
namely Aspergillus flavus, Fusarium dimerum,
Rhizopus orizae, Mucco circinelloides, Pencillium
chrysogenum, Aeromonium furcatum, Humicolo
insulans, Aspergillus niger, Trichoderma ressei and
Basidiomycetes sps were used for the study.
Agar Well Diffusion Method
Anti Bacterial Activity
The antimicrobial activity was determined
by agar well disc diffusion method. Plates for
antibacterial activity was prepared using nutrient agar
medium and sterilized, the wells were cut by using
gel puncture and previously prepared cultured
organisms were swabbed on the culture plates. A
volume of 20 µl from 5 µg, 10 µg and 15 µg of the
plant extracts were added into the well. The
assessment of antimicrobial activity was based on the
measurement of the concentration diameter of the
inhibition zone formed around the well.
Anti Fungal Activity
Potato dextrose agar was the medium used
for plate preparation in anti fungal activity, the wells
were cut by using gel puncture and the previously
prepared cultured organisms were swabbed on the
culture plates. A volume of 20 µl from 5 µg, 10 µg
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M.Thenmozhi., et al. / International Journal of Advances in Pharmaceutical Research
and 15 µg of the two plant extracts were added into
the well. The assessment of antimicrobial activity
was based on the measurement of the diameter of the
inhibition zone formed around the well.
RESULTS AND DISCUSSION
Phytochemical screening: Preliminary
phytochemical screening was performed for the plant
material and the study revealed that carbohydrates,
cardioglycosides, saponins, oils and fats, alkaloids,
flavanoids, aminoacids and quinones were present in
both plants, Eclipta alba (Table 1) whereas
Terpenoids, sterols and steroids were absent in
Eclipta alba.
Antimicrobial Activity
The antimicrobial and antifungal activity
was performed for all the five extracts (Table 2 and
3) including petroleum ether, chloroform, ethyl
acetate, methanol and water (aqueous) in the different
concentration (5, 10 and 15 µg).
The antibacterial activity was best at 15 µg
concentration in all extracts. Pseudomonas
aurugenosa was inhibited highly in Petroleum ether.
Similarly Klebsiella pneumoniae in chloroform,
Pseudomonas aurugenosa in ethyl acetate,
Salmonella typhi in methanol, Pseudomonas
aurugenosa in aqueous had shown better activity
againt the all those organisms.
Likewise antibacterial, antifungal activity
also were inhibited high in 15 µg concentration in all
extracts. Aspergillus niger was inhibited highly in
Petroleum ether. Similarly chloroform in
Trichoderma ressei, ethyl acetate in Pencillium
chrysogenum, methanol in Humicolo insulans,
aqueous in Aeromonium furcatum had shown better
activity againt the all those organism.
Eclipta alba showed a better activity in
methanol extract and the organisms, Pseudomonas
aurugenosa and Pencillium chrysogenum gave the
best activity. The second best activity was seen in
choloroform and then in petroleum ether.
Recently, much attention has been directed
towards plant extracts and biologically active
compounds isolated from popular plant species. The
use of medicinal plants play a vital role in covering
the basic health needs in developing countries and
these plants may offer a new source of antibacterial,
antifungal and antiviral agents with significant
activity against infective microorganisms 16,17 .
Methanol extract of Portulaca oleracea
showed high activity against gram positive
organisms. These might refer to the presence of
coumarins, flavanoids and saponins as chemical
components of these plants 18 which is similar to the
present study.
And in the studies described 11 the
antifungal activity of dichloromethane, methanol (80
%) and aqueous extracts obtained from Erigeron
floribundus revealed that only the dichloromethane
extract showed a good antifungal activity against the
dermatophytes and filamentous fungus used. These
studies revealed similar results to the present study.
CONCLUSION
The present study discusses on the
phytochemical and antimicrobial activities of the
plant, Eclipta alba. The phytochemicals present in
the plant were revealed and it is being assumed that
these compounds are responsible for the antibacterial
and antifungal activities present in the plant. The
specific compound responsible for the factors of
antibacterial and antifungal activities are yet to be
known and the work is in progress.
ACKNOWLEDGEMENT
We, the authors thank the Management of
Karpagam University for providing lab facilities and
constant encouragement for this research work.
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M.Thenmozhi., et al. / International Journal of Advances in Pharmaceutical Research
Table 1 Preliminary phytochemical screening of Eclipta alba
S.NO Phytochemicals Eclipta alba
1 Carbohydrates +
2
Cardioglycosides +
3
Saponins +
4 Oils and Fats +
5
Terpenoids -
6
Alkaloids +
7
Steroids and Sterols -
8 Flavanoids +
9 Tannins +
10 Phenolic compounds
+
11 Proteins +
12 Quinones +
Table 2 Antibacterial activity of Eclipta alba
Extract
Concentration
(mg/ml)
Zone of inhibition (mm) (Mean ± SD)
Petroleum
ether
Chloroform
Ethyl
acetate
Methanol
5
10
15
5
10
15
5
10
15
5
10
-
-
6 ±
0.85
8 ±
1.00
10 ±
0.85
13 ±
0.45
6 ±
0.25
9 ±
1.00
11 ±
1.52
6 ±
0.57
11 ±
-
-
-
-
-
-
-
-
-
8 ±
0.45
13 ±
6 ±
0.45
8 ±
1.02
12 ±
0.21
-
-
-
5 ±
1.00
8 ±
0.85
10 ±
0.47
6 ±
0.85
8 ±
10 ±
0.85
12 ±
1.00
17 ±
1.52
-
-
-
12 ±
0.45
14 ±
1.00
19 ±
1.00
7 ±
1.00
12 ±
6 ±
1.00
10 ±
1.00
13 ±
0.52
6 ±
0.54
9 ±
0.85
13 ±
1.00
-
-
-
6 ±
1.01
10 ±
6 ±
1.00
9 ±
1.52
12 ±
0.85
6 ±
1.00
8 ±
1.52
12 ±
0.45
11 ±
1.00
13 ±
0.85
16 ±
1.00
12 ±
1.00
14 ±
10 ±
1.00
12 ±
1.00
14 ±
1.00
-
-
-
6 ±
0.47
9 ±
0.29
12 ±
1.00
8 ±
0.52
10 ±
6 ±
0.85
8 ±
0.47
11 ±
1.00
-
-
8
±
0.56
-
-
-
8 ±
0.55
16 ±
6 ±
0.21
9 ±
0.47
12 ±
0.85
12 ±
1.00
15 ±
1.52
19 ±
0.87
-
-
-
9 ±
1.00
12 ±
-
-
9 ±
1.00
-
-
-
-
-
-
-
-
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M.Thenmozhi., et al. / International Journal of Advances in Pharmaceutical Research
Aqueous
15
5
10
15
0.21
15 ±
1.00
-
-
-
2.00
18 ±
1.76
-
-
7
±
0.45
0.76
13 ±
1.00
6 ±
0.85
7 ±
1.52
11 ±
1.00
1.00
17 ±
2.01
-
-
-
0.85
15 ±
1.00
-
-
6
±
0.76
1.00
17 ±
1.00
6 ±
1.00
12 ±
1.52
14 ±
1.00
0.85
13 ±
1.52
10 ±
1.52
12 ±
0.85
15 ±
0.82
1.00
19 ±
1.52
-
-
-
1.52
18 ±
0.85
-
-
7
±
1.00
-
-
-
-
A- Escherichia coli, B- Staphylococcus aureus, C- Pseudomonas auregenosa, D- Klebsiella
pneumoniae, E- Bacillus cereus, F- SERRATIA MARCESCENS, G- Salmonella typhi, H-
Streptococcus pyogenes, I- Proteus vulgaris, J- Citrobacter freundii,
Table 3 Antifungal activity of Eclipta alba
Extract
Concentration
(mg/ml)
Zone of inhibition (mm) (Mean ± SD)
Petroleum
ether
Chloroform
Ethyl
acetate
Methanol
Aqueous
5
10
15
5
10
15
5
10
15
5
10
15
5
10
-
-
-
6 ±
2.00
9 ±
1.00
12 ±
0.85
6 ±
1.52
8 ±
0.85
11 ±
1.00
-
-
-
-
-
-
-
-
6 ±
0.89
8 ±
0.47
11 ±
0.46
-
-
-
10 ±
1.00
18 ±
2.00
20 ±
2.00
6 ±
1.52
8 ±
1.00
11 ±
-
-
7 ±
0.85
8 ±
1.00
10 ±
1.00
14 ±
1.00
9 ±
1.00
11 ±
1.00
15 ±
1.00
8 ±
0.53
11 ±
0.85
14 ±
1.00
-
-
-
-
-
9 ±
0.89
11 ±
1.00
14 ±
1.52
6 ±
0.85
8 ±
0.45
10 ±
1.00
12 ±
1.00
14 ±
1.52
17 ±
0.85
-
-
6 ±
1.00
11 ±
1.52
13 ±
1.00
7 ±
0.99
11 ±
0.76
14 ±
0.12
7 ±
1.00
9 ±
0.85
14 ±
0.45
14 ±
1.00
16 ±
1.00
21 ±
1.00
-
-
6 ±
0.47
9 ±
0.85
11 ±
0.52
6 ±
1.00
9 ±
0.76
12 ±
1.00
6 ±
0.66
8 ±
0.78
11 ±
0.44
10 ±
0.45
12 ±
0.67
15 ±
1.00
6 ±
0.45
10 ±
1.00
14 ±
1.52
-
-
-
-
-
-
8 ±
2.00
10 ±
0.45
13 ±
1.00
8 ±
2.00
11 ±
1.52
13 ±
1.00
-
-
-
9 ±
1.00
7 ±
0.94
10 ±
2.00
14 ±
1.00
7 ±
0.33
9 ±
0.76
12 ±
1.52
12 ±
0.47
15 ±
0.67
19 ±
0.23
-
-
-
-
-
-
-
8 ±
1.00
10 ±
2.00
15 ±
0.45
8 ±
1.00
10 ±
1.52
16 ±
1.52
-
15 7 ±
6 ± - -
9 ± - - 11 ±
1.00 1.00 0.85
1.00
1.00
A- Mucco circinelloides, B- Aspergillus flavus, C- Fusarium dimerum, D- Aspergillus niger, E- Rhizopus
orizae, F- Pencillium chrysogenum, G- Aeromonium furcatum, H- Humicolo insulans, I- Trichoderma ressei,
J- Basidiomycetes
IJAPR / June 2011/ Vol. 2 / Issue. 6 / 246 - 251 250
-
-
-
-
-
-
-
-
8 ±
1.00
-
-
-
-
-
-
-
-

M.Thenmozhi., et al. / International Journal of Advances in Pharmaceutical Research
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