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Evaluation of Antioxidant And Antibacterial Activity Of Terminalia Chebula Fruit Extract

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  • Chhattisgarh Council of Science and Technology, Raipur, Chhattisgarh, India

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The present study is the continuation of a program aimed at investigation of antimicrobial properties of Terminalia chebula fruit extract to justify the traditional claim endowed upon this herbal drug as a rasayana in Ayurveda. The antimicrobial activity was evaluated according to the disk diffusion method by using Gram positive; B. subtilius, S. aureus and S. epidermidis and Gram Negative; E. coli S. flexineri, P. aeruginosa bacteria. This study show that methanolic fruits extract of Terminalia chebula Linn inhibits the growth of micro organism dose dependently. The antioxidant activity of Terminalia chebula fruit extract using Fenton reaction was also observed and we found a dose dependent inhibition of Thiobarbituric acid reactive substance (TBARS) as compared to positive control. The results of the effects of the examined Terminalia chebula fruit extract as well as control solutions on OH- radical production. They show that all extract of Terminalia chebula and control solutions as a DMSO inhibited the production of OH- radicals.These results confirm the antibacterial activity of Terminalia chebula leaves and support the traditional use of the plant in therapy of bacterial infection. These promising findings suggest the presence of antibacterial activity in the tested plant material, exhibited by its bioactive compounds, and serving them as an alternative antimicrobial agent.
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Inter. J. of Phytotherapy / Vol 5 / Issue 4 / 2015 / 160-164.
~ 160 ~
e - ISSN - 2249-7722
Print ISSN - 2249-7730
International Journal of Phytotherapy
www.phytotherapyjournal.com
EVALUATION OF ANTIOXIDANT AND ANTIBACTERIAL
ACTIVITY OF TERMINALIA CHEBULA FRUIT EXTRACT
Wasim Raja1* and Sachin Das1, 2
1Central Laboratory Facility, Chhattisgarh Council of Science and Technology, Raipur, Chhattisgarh, India.
2School of Studies in Life Science, MATS University, Raipur, Chhattisgarh, India.
INTRODUCTION
Terminalia chebula is a plant species belonging
to the genous terminalia, family combretaceae. The fruit
of the tree has been used as traditional medicine for
household remedy against various human ailments, since
antiquity. Terminalia chebula has been extensively used
in Aurveda, Unani and homoeopathic medicine and has
become a cynosure of modern medicine Terminalia
chebula exhibited antibacterial activity against a number
of bacteria species. Plant fruits appear to have evolved
complex antibiotic compounds to cure various diseases
like cancer, cardiovascular, digestive and pathogenic
bacteria.
Antibacterial activity of T. chebula extracts
against several different bacterial strains has been
reported [1-3]. The dried ripe fruits have traditionally
been used in the treatment of asthma, sore throat,
vomiting, hiccup, bleeding piles, gout, heart and bladder
diseases [4]. Its paste with water is found to be anti-
inflammatory, analgesic and having purifying and healing
capacity for wounds [5]. Its powder is used as an
astringent in loose, bleeding gums and oral ulcers. It is
used to increase the appetite, liver stimulant, as
stomachic, as gastrointestinal prokinetic agent, in chronic
diarrhea and mild laxative [6].
Corresponding Author:- Wasim Raja Email: drwasimraja84@gmail.com
ABSTRACT
The present study is the continuation of a program aimed at investigation of antimicrobial properties of
Terminalia chebula fruit extract to justify the traditional claim endowed upon this herbal drug as a rasayana in
Ayurveda. The antimicrobial activity was evaluated according to the disk diffusion method by using Gram positive;
B. subtilius, S. aureus and S. epidermidis and Gram Negative; E. coli S. flexineri, P. aeruginosa bacteria. This study
show that methanolic fruits extract of Terminalia chebula Linn inhibits the growth of micro organism dose
dependently. The antioxidant activity of Terminalia chebula fruit extract using Fenton reaction was also observed
and we found a dose dependent inhibition of Thiobarbituric acid reactive substance (TBARS) as compared to
positive control. The results of the effects of the examined Terminalia chebula fruit extract as well as control
solutions on OH- radical production. They show that all extract of Terminalia chebula and control solutions as a
DMSO inhibited the production of OH- radicals.These results confirm the antibacterial activity of Terminalia
chebula leaves and support the traditional use of the plant in therapy of bacterial infection. These promising findings
suggest the presence of antibacterial activity in the tested plant material, exhibited by its bioactive compounds, and
serving them as an alternative antimicrobial agent.
Key words: Terminalia chebula, Antioxidant, Antimicrobial activity, Disk diffusion, Inhibition zone.
Inter. J. of Phytotherapy / Vol 5 / Issue 4 / 2015 / 160-164.
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It is effective in inhibiting Helicobactor pylori
[1], Xanthomonas campestris pv. citri [7] and Salmonella
typhi. An aqueous extract of T. chebula fruit exhibits
antifungal activity against a number of dermatophytes and
yeasts [8]. Being a mild laxative, it is mild herbal colons
cleanse [9]. It is used in various weaknesses, nervous
irritability. It promotes the receiving power of five senses
[10]. Antibacterial activities of T. chebula extracts against
several bacterial strains have been reported [11]. In view
of these reported medicinal values, the present work was
carried out to examine the antibacterial potential of an
ethanol extract of T. chebula fruits against clinically
important CLSI (Clinical and Laboratory Standards
Institute) reference bacterial strains. Terminalia chebula
is routinely used as traditional medicine in the name of
‘Kadukkaai’ by tribal of Tamil Nadu in India to cure
several ailments such as fever, cough, diarrhea,
gastroenteritis, skin diseases, candidiasis, urinary tract
infection and wound infections16. Antibacterial activity
of Terminalia chebula extracts against several bacterial
strains has been reported. Extracts from different parts of
diverse species of plants like root, flower, leaves, seeds,
etc. exhibit antibacterial properties were applied on cotton
material for wound, healthcare care application. It is a
well known fact that the demand for the herbal drug
treatment of various ailments is increasing and plant drugs
from the ayurvedic system are being explored more, not
only in India but also globally. As a result, many research
studies are being undertaken and there is a need for an
update and to put them together. In this article an attempt
has taken to recapitulate available pharmacological
studies for Terminalia chebula. The first part of the article
explains the natural species identy, habitat, botanical
description, chemical constituents. To the best of our
knowledge, there have been no published reports
concerning the antimicrobial activity of aqueous extract
of T. chebula fruits against B. subtilus, S. aureous,
S.epidermis, E.coli, S. flexineria and P.auriginosa and
phytochemical activity of this plant. We have, therefore
focused on its antimicrobial activity and phytochemical
analysis in our study.
MATERIALS AND METHOD
Extract preparation
Terminalia chebula Fruits (100 g) was defatted
with petroleum ether (1000 ml) and the residue was
extracted in 50% methanol with the help of soxhlet
extraction unit. The sample was collected and
concentrated in water bath at 40-50oC and dried in hot air
oven at 40oC. The dried powder was kept in air tied box.
Microorganism
The test organism included the gram positive
bacteria; Bacillus cereus (ATCC 11778), Staphylococcus
aureus (ATCC 25923), and gram negative bacteria;
Klebsiella pneumoniae (NCIM 2719), Escherichia coli
(ATCC 25922) and Pseudomonas pseudoalcaligenes
(ATCC 17440). All the bacterial strain were obtained
from National Chemical Laboratory (NCL), Pune, india.
The bacteria were grown in the nutrient broth at 37oC and
maintained on nutrient agar slant at 4oC.
In vitro Antioxidant activity:
The Fenton reaction was used to generate free
radicals in a test system and the free radical scavenging
activity was determined by the degradation of
deoxyribose as standardized by Elizabeth and Rao (1990)
[12]. Fe3+, ascorbate, EDTA, H2O2 in the system
produced hydroxyl redical which react with deoxyribose
and set off a series of reaction that the result is generation
of Thiobarbituric acid reactive substance (TBARS). The
measurement of TBARS thus gives and index of free
radical activity. Radicals scavenging by protectors will
result in the inhibition of TBARS. This was showed its
antioxidant activity.
Antibacterial Assay: Antibacterial activity of terminalia
chebula fruits extract was determined by agar disk
diffusion method at four different concentrations i.e., 100,
75, 50 and 25 mg/ml. Muller Hinton agar was prepared
according to the manufactur’s instruction and the plates
were seeded with appropriate micro organism (bacillus
cereus, staphylococcus aureus and Klebsiella pneumonia,
Escherichia coli and pseudomonas pseudoalcaligenes).
Discs of 6 mm diameter were prepared from Whatmann
filter paper No. 1 and sterilized. The discs were than
impregnated with the extracts and solvent DMSO.
Antibiotics for Gram positive (TE- Tetracycline, OF-
Ofloxacin, AZ- Azithromucin and PC- Pipracillin) and
Gram negative (Fu- Nitrofurantoin, GM- Gentamicine,
CX- Cefotaxime and NF- Norfloxaci,5 µl/disc) bacteria
were used as standard. The plates were incubated at 37oC
for 24 hrs and the zone of inhibition was measured with
measuring scale. This experiment was carried out in
triplicate for their confirmation.
RESULT
Antioxidant Activity
The results of the effects of the examined
Terminalia chebula fruit extract as well as control
solutions on OH- radical production. They show that all
extract of Tinosporia cardifolia and control solutions as a
DMSO inhibited the production of OH- radicals. The %
of free racial scavenging activity of hydro-methanolic
extract of Terminalia chebula presented in Table 1 have
reducing power, the free radial OH- scavenging activity
of the extract increases with increasing the concentration.
Antimicrobial Activity
The initiation of microbial growth was
considered as zero hour and further accordingly reading
was taken. Our present study shows that antibacterial
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activity of 50 % methanolic extract of Terminalia chebula
against S. epidemidis is best in 100 % concentration after
12 hrs. (08.90 mm zone of inhibition). Although 75%
concentration is having mild effect as 08.00 mm zone of
inhibition. In S. aureus 100% concentration of extract is
having good antibacterial activity at maximum zone of
inhibition 09.00 mm. On the other hand 75 % is showing
static activity from, with zone of inhibition of 06.00 mm.
For E. coli 100% concentration of extract show maximum
zone of inhibition 14.20 mm. Although the same effect of
75% concentration of extract is also revealing as showing
zone of inhibition 10.10 mm. In the case of Sh. flexineri
75% and 100% concentration of extract show good
activity with zone of inhibition of 08.20 mm and 07.50
mm respectively. The above observations suggest that
different concentration (50 %, 75 % & 100 %) were
having good antibacterial activity against B. subtilius, S.
aureus, S. epidermidis, E. coli, Sh. Flexineri, Ps.
aeriuoginosa. Thus the extract is showing varying activity
against all microorganisms. On comparing the zone of
inhibition of extract to that of standard antibiotics extract
showed better activity than Tetracyclin and Azithromycin.
But extract is not potent than Ofloxacin and Cefotaxime
in these conditions (Table 2 and 3).
Figure 1. Antioxidant activities of Terminalia chebula
fruit extract using Fenton reaction
Figure 2. The study of anti-bacterial activities of T.
chebula extracts using Gram Negative
Figure 3. The study of anti-bacterial activities of T.
chebula extracts using Gram Positive
Figure 4. Antibacterial Activity of Standard Antibiotic
Table 1. Antioxidant activities of Terminalia chebula fruit extract using Fenton reaction
Constrictions (in μl)
% Inhibition of TBRAS
Terminalia chebula
10
17.00±0.93
20
22.54±1.33
30
37.73±1.69
40
52.33±1.50
50
66.22±1.63
Blank 0.4320
Inter. J. of Phytotherapy / Vol 5 / Issue 4 / 2015 / 160-164.
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Table 2. The study of anti-bacterial activities of standard antibiotics using disk diffusion method
Table 3. The study of anti-bacterial activities of T. chebula extracts using Disk Diffusion method (Mean + SE).
DISCUSSION
The present study was conducted to study the
antioxidant and antibacterial activity of Terminalia
chebula used by Indian peoples to show that therapeutic
properties. The antibacterial activity was expressed at
varying degrees with the activity being both strain and
dose dependent. Six bacteria’s were used for antibacterial
studies. Medicinal plants are being used by large
proportion of Indian population. It has also been widely
observed and accepted that the medicine value of plant
lies in the bioactive phytochemicals present in the plants.
The antibacterial study suggest that different
concentration (50 %, 75 % & 100 %) of plant extract were
having good antibacterial activity against B. subtilius, S.
aureus, S. epidermidis, E. coli, Sh. Flexineri, Ps.
aeriuoginosa. Thus the extract is showing varying activity
against all microorganisms. On comparing the zone of
inhibition of extract to that of standard antibiotics extract
showed better activity than Tetracyclin and Azithromycin.
But extract is not potent than Ofloxacin and Cefotaxime
in these conditions. This finding is interesting, because in
the traditional method of treating a microbial infection,
decoction of the plant parts or boiling the plant in water
was employed. Whereas, according to the present study,
preparing an extract with an organic solvent (acetone and
ethanol) shows a better antimicrobial activity [13-14].
It has been known that superoxide dismutase SOD is a
metalloproteinase that is involved in the antioxidant
defense mechanism, which plays an important role in the
protection of cells against reactive oxygen system (ROS)
by lowering the steady state of superoxide anions. Also,
SOD converted superoxide radical to hydrogen peroxide
and molecular oxygen which in turn can be counteracted
by catalase or glutathione peroxidase reaction thereby
reducing the level of cellular damage [15].
Another set of experiment the effects of the
examined Terminalia chebula fruit extract as well as
control solutions on OH- radical production. They show
that all extract of Tinosporia cardifolia and control
solutions as a DMSO inhibited the production of OH-
radicals. The % of free racial scavenging activity of
hydro-methanolic extract of Terminalia chebula presented
in Table 1, 2 and 3 have reducing power, the free radial
OH- scavenging activity of the extract increases with
increasing the concentration.
The present study was conducted to study the
antibacterial activity of Terminalia chebula used by
Indian peoples to show that therapeutic properties. The
antibacterial activity was expressed at varying degrees
with the activity being both strain and dose dependent six
bacteria’s were used for antibacterial studies medicinal
plant are being used by large proportion of Indian
population. It has also been widely observed and accepted
that the medicinal value of plant lies in the bioactive
phytochemical present in the plants. The present
investigation suggested that possible mechanism may be
due to free radical scavenging activities which may be
due to presence of in the extract.
Hence, it is concluded that Terminalia chebula
extract has an alternative medicine for various disease. As
the global scenario is now changing towards the use of
nontoxic plant products having traditional medicinal use,
development of modern drugs from T. chebula should be
emphasized for the control of different disease. Further
the plant is used in the treatment of gastric ulcer,
Sl
Bacterial
Stain
Bacteria Use
Zone of Inhibition (In MM)
25%
50%
75%
100%
1
Gram
Negative
(-)
E. coli
07.00 ±2. 78
9.20 ± 1.28
10.10 ± 1.64
14.20 ±1.77
Ps. Aeriuoginosa
07.20 ± 1.27
08.00 ± 5.77
06.50 ± 1.40
09.20 ±0.63
Sh. Flexineri
06.66 ±1.27
09.00 ± 1.54
07.50 ±11.54
08.20 ± 1.55
2
Gram
Positive
(+)
Bacillus subtilis
07.00 ± 1.46
08.20 ± 1.40
07.00 ± 7.80
08.00 ± 1.60
S. epidermidis
7.50 ± 1.27
05.90 ± 1.77
08.00 ± 5.78
08.90 ± 1.27
S. aureus
08.00 ± 2.77
07.20 ± 2.40
06.00 ± 5.77
09.00 ± 1.54
Sl
Name of Bacteria
Zone of Inhibition (In MM)
1
Gram Negative (-)
TE10
OF5
AZ15
PC5
E.coli
17.00
15.00
14.00
11.00
Ps.aeriuoginosa
15.00
17.00
19.00
16.00
Sh. flexineri
12.00
12.00
17.00
12.00
2.
Gram Positive (+)
FU10
GM5
CX15
NF5
Bacillus subtilis
11.00
12.00
09.00
14.00
S. aureus
16.00
14.00
19.00
17.00
S. epidermidis
16.00
16.00
14.00
18.00
Inter. J. of Phytotherapy / Vol 5 / Issue 4 / 2015 / 160-164.
~ 164 ~
constipation, general debility, piles. Hence, this plant
provides a significant role in the prevention and treatment
of a disease. Further evaluation needs to be carried out in
order to explore the concealed areas and their practical
clinical applications, which can be used for the welfare of
the mankind.
ACKNOWLEDGMENT
The authors are thankful to Prof. M.M.
Hambarde, Director General, Chhattisgarh Council of
Science and Technology, Raipur (Chhattisgarh) India, for
providing facility and technical support to carry out the
above work.
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ResearchGate has not been able to resolve any citations for this publication.
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Antibacterial activity of black myrobalan (Fruit of Terminalia chebula Retz) against uropathogen Escherichia coli
  • R R Chatopadhyay
  • S K Bhattacharyya
  • C Medda
  • Chanda S Datta
  • S Pal
Chatopadhyay RR, Bhattacharyya SK, Medda C, Chanda S, Datta S, Pal NK. Antibacterial activity of black myrobalan (Fruit of Terminalia chebula Retz) against uropathogen Escherichia coli. Phcog. Mag, 11, 2009, 212-15.