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Document heading doi:10.1016/S2222-1808(14)60619-8 襃 2015 by the Asian Pacific Journal of Tropical Disease. All rights reserved.
Evaluation of antioxidant and antimicrobial potential of different leaves
crude extracts of Omani Ficus carica against food borne pathogenic
bacteria
Afaf Mohammed Weli, Afaf Ali Mohammed Al-Blushi, Mohammad Amzad Hossain*
School of Pharmacy, College of Pharmacy and Nursing, University of Nizwa, P. O. Box 33, Postal Code 616, Nizwa, Sultanate of Oman
Asian Pac J Trop Dis 2015; 5(1): 13-16
Asian Pacific Journal of Tropical Disease
journal homepage: www.elsevier.com/locate/apjtd
*Corresponding author: Dr. Mohammad A. Hossain, School of Pharmacy, College
of Pharmacy and Nursing, University of Nizwa, P. O. Box 33, Postal Code 616, Nizwa,
Sultanate of Oman.
Tel: +96892327578
Fax: +96892877745
E-mail: hossainabi@gmail.com
Foundation Project: Supported by University of Nizwa, Nizwa, Sultanate of Oman
(Grant No. 507/SOP/OB/1/2013).
1. Introduction
Ficus carica (F. carica) is a medicinal plant constituting
one of the largest genera with about 750 species. It is a woody
plants, trees and shrubs. Primarily it is found in subtropical
and tropical regions throughout the world[1]. The genus is
remarkable for the large variation in the habits of its species.
It is commonly referred as fig. F. carica grows well up to a
height of 6.9-10 m, with smooth grey bark. It is well known for
its large and fragrant leaves. The leaves are 12-25 cm long and
10-18 cm across, and deeply lobed with three or five lobes[2-4].
The complex inflorescence of the common fig consists of a
hollow fleshy structure called the syconium, which is lined
with numerous unisexual flowers. The edible fig fruit is the
mature syconium on the outside and numerous one-seeded
fruits on the inside. The fruit is 3-5 cm long, with a green skin,
sometimes ripening towards purple or brown. F. carica has
milky sap. It is rich in vitamins, mineral elements, water, and
fats. Figs are one of the highest plant sources of calcium and
fiber[5]. The chemical constituents in the leaves of F. carica
PEE R REVIEW ABSTR ACT
KEYWORDS
Ficus carica, Al-Teen, Maceration method, DPPH, Antioxidant, Antimicrobial potential
Objective: To prepare different polarities crude extract from the leaves of Ficus carica and to
evaluate their antioxidant and antimicrobial potential against food borne pathogenic bacterial
strains.
Methods: The dried leaves were macerated in absolute ethanol for one week. The ethanol
was evaporated and the crude extract was defatted with ethanol-water. The defatted hydro
alcoholic crude extract was successively extracted with hexane, chloroform and ethyl acetate.
The antioxidant potential was determined against 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay.
Evaluation of antimicrobial potential of different crude extracts against selected Gram positive
and Gram negative bacteria by agar disc diffusion method.
Results: The total extraction yield was 2.2%. The highest extraction yield was in chloroform and
the lowest in hexane. The antioxidant results were found in the order of hydro alcoholic>ethyl
acetate>hexane>chloroform. Hydro alcoholic crude extract and its derived fractions display
moderate antimicrobial potential against the selected bacterial strains such as Staphylococcus
aureus, Escheichia coli and Pseudomonas, in the range of 0%-13%.
Conclusions: It is concluded that the hydro alcoholic and ethyl acetate crude extracts of Ficus
carica possess very good antioxidant and antimicrobial potential.
Contents lists available at ScienceDirect
Peer reviewer
Prof. Dr. Huge Dougle, Institute of
Natural Products, University of Yale,
UK.
E-mail: hugedu@gmail.com
Comments
The present study on antioxidant
and antimicrobial activity of various
leaves crude extracts of F. carica is
giving the valuable brief and scientific
information about this plant.
Details on Page 16
Article history:
Received 8 Apr 2014
Received in revised form 18 Apr, 2nd revised form 6 May, 3rd revised form 15 May 2014
Accepted 12 Jun 2014
Available online 11 Jul 2014
Afaf Mohammed Weli et al./Asian Pac J Trop Dis 2015; 5(1): 13-16
14
were protein (67.6%), fat (4.3%), fiber (1.7%), total ash (4.7%),
nitrogen free compounds (5.3%), pentoses (16.4%); carotene
(3.6%), bergaptene, stigmasterol, sitosterol, and tyrosine[6]. The
sap of the fig’s green parts is an irritant to human skin[5]. All
parts of this plant such as bark, leaves, tender shoots, fruits,
seeds, and latex are medicinally important[2]. The fig is a very
nourishing food and used in industrial products. The leaves
of this plant have anti-diabetic properties and reduce the
amount of insulin needed by diabetics. The leaves have also
the ability to inhibit the growth of cancer cells and to prevent
colon cancer[7]. Figs are a good source of potassium and it is
very important mineral to help control blood pressure[8]. The
fiber of figs also helps to reduce weight and is recommended
for obese people[8]. The ethanol crude extract of F. carica
at doses of 100, 200 and 300 mg/kg showed significant dose-
dependent reduction in normal body temperature and yeast
provoked elevated temperature. Its crude extracts showed
high acute toxicity with hemorrhagic enteritis. In addition,
the crude extracts showed a weak anthelmintic efficacy. The
plant crude extracts and their mixture decreased the level of
mutations induced by N-methyl-N’-nitro-N-nitrosoguanidine
in viciafaba cells, demonstrating the ability to decrease the
genotoxicity of environmental mutagens[9]. The main objective
of the present study was to determine the antioxidant and
antimicrobial potential of different concentrations and different
polarities of leaves crude extracts of F. carica against selected
food borne pathogenic bacterial strains such as Staphylococcus
aureus (S. aureus), Escheichia coli (E. coli) and Pseudomonas.
2. Materials and methods
2.1. Chemicals
The chemicals such as ethanol, chloroform, ethyl acetate,
methanol and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were
obtained from Sigma-Aldrich Chemical Company Limited. The
food borne pathogenic bacterial strains S. aureus, E. coli and
Pseudomonas aeruginosa (P. aeruginosa) were obtained from
Microbiology Laboratory, Department of Biological Sciences,
College of Arts and Sciences, Nizwa University, Sultanate of
Oman. Filter paper discs of diameter 5 mm were obtained from
Whatman Company. Nutrient agar and plastic Petri dishes were
purchased from Sharlau Chemie Company. Deionized water
was used throughout the experiment. Shimadzu1800 UV-visible
spectrophotometer was used for analysis.
2.2. Plant samples
The leaves of F. carica sample were collected from Izki on
23 October, 2012 in the afternoon at 4.00-6.00 pm. The collected
leaves samples were transported to the lab for processing.
2.3. Preparation of crude extracts
The whole leaves samples were separated from the affected
one and washed with water. The fresh leaves samples were
placed on newspapers and dried under shade. After complete
drying, the leaves were grinded using kitchen grinder. The
powdered leaves samples (349.31 g) were taken in a three
liter beaker and added 95% ethanol (1.5 L) for one week. After
the complete extraction, the solvent was decanted out and
filtered under vacuum using Buchner apparatus to give clear
solution. The ethanol was evaporated at low pressure using
rotary evaporator to obtain crude ethanol extract. The crude
extract was defatted with water and extracted successively with
hexane, chloroform and ethyl acetate.
2.4. Radical scavenging potential by DPPH method
Free radical scavenging potential of different crude extracts
was estimated as described by Blois[10,11]. Four concentrations
(12.5, 25, 50, 100 µg/mL) were prepared from different crude
extracts such as hexane, chloroform, ethyl acetate and hyrdo
alcoholic. Four milliliter of each concentration were placed in
the separate test tube. One milliliter of freshly prepared DPPH
solution was added and shaken vigorously. After that, all the
test tubes were placed at room temperature in dark place for 45
min. The control was prepared in the same way without adding
any crude extract. The absorption of the samples was measured
using UV spectroscopy at 517 nm. The inhibition percentage
was calculated using the formula:
% Inhibition = Acontrol - Aextract
Acontrol
伊 100
2.5. Antibacterial assay
The evaluation of antibacterial test was carried out by
the agar disc diffusion method[12]. Four concentrations of
each extract was prepared using serial dilution method with
dimethyl sulphoxide to obtained 2 000, 1 000, 500, and 250 µg/
mL solution. Filter paper discs were macerated with each
concentration and placed on previously prepared agar gel
plate. All the plates were incubated with microorganism at
37 °C for 24 h. Amoxicillin was used as a positive control.
The calculation of antibacterial activity was determined by
measuring the diameter of the zone of inhibition against the
tested food borne pathogenic bacterial strains.
3. Results
3.1. Crude extracts from the leaves of F. carica
The powdered leaves samples were extracted with ethanol for
one week. The ethanol was evaporated at low pressure using
rotary evaporator to obtain crude ethanol extract. The crude
extract was defatted with water and extracted successively with
hexane, chloroform and ethyl acetate. The total yield was 2.7%.
The highest extraction yield was in chloroform and the lowest
in hexane and the order was chloroform>ethyl acetate>hydro
alcoholic>hexane.
3.2. Antioxidant potential
The antioxidant potential was determined by agar gel
diffusion method. The results of antioxidant potential for
Afaf Mohammed Weli et al./Asian Pac J Trop Dis 2015; 5(1): 13-16 15
hexane, chloroform, ethyl acetate and hydro alcoholic extract
against DPPH radical are shown in Figure 1. All crude extracts
from F. carica inhibited the DPPH radical. Hydro alcoholic
crude extract showed radical scavenging of more than 90%
at all concentrations. The lowest inhibition was shown by
chloroform.
120.00
100.00
80.00
60.00
40.00
20.00
0.00
12.5 25 50 100 200
Hexane Chloroform Ethyl acetate Hydro alcholic
Figure 1. Antioxidant potential of different crude extracts against DPPH.
%
inhibition
Concentration (µg/mL)
3.3. Antimicrobial potential
The determination of antibacterial potential of hexane, ethyl
acetate, chloroform, hydro alcoholic crude extracts of F. carica
against three food borne pathogenic bacterial strains were
calculated the presence or absence of inhibition zones. The
exhibition of antibacterial potential of four crude extracts of F.
carica were shown against S. aureus, E. coli and P. aeruginosa
bacterial strains at the concentrations of 2 000, 1 000, 500 and
250 µg/mL with dimethyl sulphoxide. Almost all crude extracts
of F. carica were showed moderate potential of antibacterial
activity against E. coli, P. aeruginosa and S. aureus bacteria at
the concentrations of 2 000, 1 000, 500 and 250 µg/mL (Figure 2).
Amoxicillin was used as a positive control.
2000 1000
Hexane
14
12
10
8
6
4
2
0
Chloroform Ethyl acetate Hydro alcoholic
Concentration (µg/mL)
E. coli S. aureus P. aeroginosa C. albicans
1000 1000 10002000 2000 2000
500 500 500 500 250250 250
Figure 2. Antimicrobial potential of different leaves crude extracts of F.
carica against E. coli, S. aureus and P. aeroginosa.
Inhibition zones (mm)
4. Discussion
The powdered leaves samples were extracted with ethanol
and defatted with water and extracted successively with
hexane, chloroform and ethyl acetate. The total extraction yield
was 2.7%. The highest yield was obtained from chloroform and
lowest from hexane. This result indicated that the chloroform
crude extract contained high percentage of non and semi
polar organic compounds. The antioxidant potential was
determined by well established DPPH method. The principle
of DPPH method based on production of free radical[13]. The
hydro alcoholic crude extracts produced more free radical and
chloroform crude extracts produced less free radical. Therefore,
the highest antioxidant potential was obtained hydro alcoholic
crude extract among the other crude extracts and the lowest
was chloroform. The antioxidant results was found in the order
of hydro alcoholic>ethyl acetate>hexane>chloroform (Figure 1).
The hydro alcoholic extract showed highest potential and the
ethyl acetate also showed high potential compared to hexane
and chloroform crude extracts. The variation of antioxidant
potential might be poly phenolic chemical compounds in
the crude extracts[14,15]. It was observed to have the highest
antioxidant potential in DPPH assay, which is in agreement with
previous study[11].
There are several studies of antimicrobial potential of
F. carica leaves crude extracts. Jung reported that the
methanol crude extract from the leaves of F. carica exhibited
strong potential against E. coli but weak potential against
S. aureus[14]. Another good study demonstrated that the
variation of antimicrobial potential was due to some flavonoid
compounds in the leaves of F. carica[15]. Ahmad et al. reported
that antimicrobial potential of methanol leaves crude extract
of F. carica against five bacterial strain Bacillus cereus,
Enterobacter aerogens, Klebsiella pneumoniae, Bacillus
subtilis, Staphylococcus epidermidis at different concentrations
was found in the following decreasing order Staphylococcus
epidermidis>Klebsiella pneumoniae>Bacillus subtilis>Bacillus
cereus>Enterobacter aerogens[16]. The antibacterial potential by
agar disc diffusion assay showed that methanol crude extract
of F. carica exhibited potential against pathogenic as well as
non-pathogenic test bacteria. The authors also mentioned that
significant effect on growth inhibition of Gram positive and
Gram negative bacteria[16].
In this present study, highest strong potential was obtained
from ethyl acetate against E. coli and P. aeroginosa at all
applied concentration but exhibited moderate potential against
S. aureus at the concentration 2 000 and 1 000 µg/mL (Figure
2). However, the concentration at 500 and 250 µg/mL did not
show any microbial potential against S. aureus. Chloroform
crude extracts showed moderate potential against E. coli at
all applied concentration but the other pathogenic bacterial
strains showed moderate potential only at the concentrations of
2 000 and 1 000 µg/mL. However, 500 and 250 µg/mL did not show
any activity against S. aureus and P. aeroginosa. Hexane crude
extract showed moderate potential against all applied bacteria
at the concentration of 2 000, 1 000, and 500 µg/mL. Hydro
alcoholic extract also showed potential against all pathogenic
bacteria at concentration 2 000 and 1 000 µg/mL.
In conclusion, the leaves of F. carica were found to possess
strong antioxidant and moderate antimicrobial activity. Gram
negative bacteria were found to be more susceptible than
Gram positive bacteria indicating that active ingredients
in the studied extracts are inhibiting growth of bacteria via
unusual mechanism. It will be thus interesting to isolate
these compounds and further investigated their antimicrobial
properties. Antioxidant compounds are known to possess both
anticancer and neuro protective characteristics. Hence, it is
suggested to extend phytochemical investigation of F. carica
Afaf Mohammed Weli et al./Asian Pac J Trop Dis 2015; 5(1): 13-16
16
from Oman in order to evaluate further its pharmacological
potentials.
Conflict of interest statement
We declare that we have no conflict of interest.
Acknowledgements
The authors are grateful to Prof. Dr. Nafsiah Binti Shamsudin,
Dean, College of Pharmacy and Nursing, University of Nizwa,
Sultanate of Oman for her continuous encouragement during
the work and all laboratory facilities. The authors are also
grateful to University of Nizwa, Nizwa, Sultanate of Oman
for providing all chemicals and other expenses from their
internal fund to carry out this project (Grant No. 507/SOP/
OB/1/2013). Thanks also go to Qasim Al-Riyami, Assistant
Dean, Training, School of Pharmacy, University of Nizwa for
his continuous encouragement and Khaloud Ali Said Al-Alawi
and Ahlam Rashed Alabri, Lab Technicians, Natural Product
Lab, University of Nizwa for their continuous help during the
experiment. The authors wish to express sincere gratitude to
the Central Instrument Laboratory, College of Agriculture and
Marine Sciences, Sultan Qaboos University, Sultanate of Oman
where the tests were confirmed.
Comments
Background
F. carica plant is a medicinal plant constituting one of the
largest genera with about 750 species. All parts of this plant
such as bark, leaves, tender shoots, fruits, seeds, and latex are
medicinally important. The fig is a very nourishing food and
used in industrial products. The leaves of this plant have anti-
diabetic properties and reduce the amount of insulin needed
by diabetics.
Research frontiers
The aim of this study is to prepare various crude extracts
using different polarities of solvent and to quantitatively
evaluate antioxidant activity and antimicrobial activity of F.
carica collected from Izki, Nizwa, Sultanate of Oman.
Related reports
According to the literature search, no work has been done
on Omani F. carica by the researcher. The other parameters of
this plant have been done by other researchers.
Innovations & breakthroughs
Although the experimental work done by the author is routine
work, it gives the new information and data to the scientific
community.
Applications
This plant is used worldwide as a traditional herbal
medicine. According to the paper, there are so many bioactive
compounds that can be used to prepare medicine.
Peer review
The present study on antioxidant and antimicrobial activity of
various leaves crude extracts of F. carica is giving the valuable
brief and scientific information about this plant.
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