Plant Tissue Culture and Bioprospecting Laboratory, M.S. Swaminathan Research
Cross Street, Taramani, Chennai- 600113, India
Community Agrobiodiversity Centre, M.S. Swaminathan Research
Foundation, Wayanad- 673121, Kerala, India
The study was carried out on antimicrobial, antioxidant and anticancer activity of Syzygium
of 24 compounds were identified among which 4-(4-ethylcyclohexyl)-1-pentyl-Cyclohexene (24.07 %) and
Linoleic acid (
Antimicrobial activity of the leaf was observed on six
bacterial and three fungal species, whose MIC values ranged from 100 to 500
g/ml. At higher concentration,
observed that the leaf sample were able to effectively inhibit the growth of Hep 2 cells.
belonging to family Myrtaceae comprising of
Africa, subtropical and tropical Asia, Australia,
New Caledonia, New Zealand, Pacific island;
among these 80 species have been reported from
and more than 75 species from India
Syzygium benthamianum is one of the species
that has been categorized as vulnerable tree
species under the IUCN
red list of threatened
and growth inhibitory effects against
. Syzygium species are also found
, Antinociceptive activity
Till date there is no literature available on the
Therefore, the present study was undertaken to
analyze some of the medicinal properties of
Syzygium benthamianum leaf extract constituents
and their biological activities.
The plant material was collected from
*Corresponding author (Palanisami Eganathan)
2011; accepted in revised form 28
Kerala, India, and identified as Syzygium
Centre (CAbC), M.S. Swaminathan Research
Foundation, Wayanad, Kerala and voucher
specimens deposited in the herbarium. The shade
dried leaves were powdered and subjected to
solvent extraction. Compound extraction was
carried out using soxhlet apparatus
collected and evaporated using rotary evaporator
at 77°C. The dried extracts were dissolved in
dimethyl sulphoxide (DMSO) and evaluated for
their efficacy against micro organisms.
Gas chromatography-Mass spectrometry
The GC (Agilent 6890) conditions were as
follows: DB-5 column (30 m X 0.25 mm X
temperatures were maintained at 250°C in the
injector and detector, helium was used as carrier
gas at a flow rate of 1 ml/min; oven temperature
was initially maintained at 100°C for 5 min and
then raised to 220°C at a rate of 10°C/min and
held at 220°C for 18 min. The MS (Agilent 5973
inert MSD) electron multiplier 2188.2 V, mass
spectra data were acquired in the scan mode in
m/z range 58-550. The compounds of the leaf
extract were identified by comprising their
retention indices (RI), with those on the stored
in NIST (National Institute of Standards and
Technology) library and by comparing their mass
spectra with the data already available in
three fungal (Aspergillus niger, Alternaria
were used in this study. The bacterial stock
cultures were maintained on nutrient agar
medium and fungal culture on potato dextrose
agar medium, and were stored at 4°C.
Determination of antimicrobial activity
The extracts were tested for their antimicrobial
activity by Disc diffusion method. Bacterial
species were sub-cultured on nutrient agar
medium and fungal species on potato dextrose
agar medium, which were then incubated at
37°C for 24 h and 27°C for 48 h respectively.
The test solutions of the dried extracts at the
concentrations of 1000
μg, 500 μg, 250 μg, 100
μg/ml were impregnated on sterile discs.
Streptomycin and Nystatin were used as positive
controls. The disc impregnated with ethyl
acetate was used as negative control. The discs
were placed on the surface of the nutrient agar
for bacteria and incubated at 37°C for 24 h. The
discs were placed on the surface of potato
dextrose agar for fungi and incubated at 27°C
for 48 h. Inhibition zones were calculated as the
difference between disc diameter (6mm) and the
diameters of inhibition
. The antibacterial
of minimum inhibitory concentration and
minimum lethal concentration by micro broth
Determination of antioxidant activity with
The free radical scavenging activity of the
sample was evaluated using DPPH
. The sample
using methanol. Pure methanol was taken as blank
and 0.016 % butylated hydroxyl toluene (BHT)
was taken as the standard. 2.7 ml of methanol,
μl of sample and 200 μl of DPPH reagent
kept in dark incubation at RT for 30 mins.
Samples were visualized in UV-VIS spectro-
photometer at wavelength of 517nm.
Percentage of DPPH radical scavenging activity
of the sample was calculated as:
is the absorbance of the solution
level and A
is the absorbance of the DPPH
The anticancer activity of the sample was
measured using 3-(4,5-dimethyl thiazol-2yl)-2,5-
diphenyl tetrazolium bromide (MTT) assay
The monolayer culture of Hep2 cells at a
24 well titre plates. Cells were permitted to adhere
for 24 h, and then treated with different dilution
(1:1 to 1:64) of the extract for 24 h; 200
MTT (5 mg/ml in PBS) were added to each well,
in 5 % CO
incubator. After removal of the
The absorbance was recorded at the wavelength
of 570 nm. The effect of the extracts on cell
growth inhibition was assessed as percent cell
viability, where vehicle-treated cells were taken
as 100 % viable.
Percentage of viable cell concentration was
Viability (%) = (Optical Density of sample x
Crude extract of leaf showed a total of 24
compounds among which 4-(4-ethyl-
cyclohexyl)-1-pentyl-Cyclohexene (24.07 %)
Linoleic acid (
taene (10.27 %), 9,17-Octadecadienal,(z)- (9.96
Z,E-3,13-Octadecadien-1-ol (7.14 %)
tuents (Table 1).
Compounds are listed and elution from DB5 column
Values calculated from published literature (DB5 column)
tested bacterial isolates ranged from 100 to 500
g/ml inhibits the growth of Proteus vulgaris and
concentration of 500
g/ml. All other microbial
inhibitory concentration at 250
The ethyl acetate extract of Syzygium
potent free radical scavengers in comparison
with BHT, a commercial antioxidant. At higher
g/ml) the extract has
scavenging activity (Table 3). Syzygium
activity with Syzygium cumini fruit
on cell viability of Hep2 cells was used. The ethyl
acetate extract treatment to these cell lines
resulted in a remarkable dose-dependent inhi-
bition of cell growth. The extract showed maxi-
mum cell inhibition at higher concentration
(Table 4). The extract of Syzygium benthamianum
showed higher activity on cancer cell lines and
this result correlated with the activity exhibited
by Syzygium cumini on AML cells
Thus from the present study it was proven that
effective biological properties against pathogens
and cancer cells. In addition also it exhibits high
scavenging activity against free radical
comparable with that of standard available drugs.
Authors are thanking to the Department of
Biotechnology, GOI, New Delhi for financial
78.08 ± 1.65
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