IJRPC 2015, 5(4), 753-763 Madhavi et al.
ISSN: 2231
2781
753
INTERNATIONAL JOURNAL OF RESEARCH IN PHARMACY AND CHEMISTRY
Available online at
www.ijrpc.com
PHYTOCHEMICAL SCREENING AND EVALUATION
OF BIOLOGICAL ACTIVITY OF ROOT EXTRACTS
OF
SYZYGIUM SAMARANGENSE
M. Madhavi* and M. Raghu Ram
Department of Botany & Microbiology, Acharya Nagarjuna University,
Guntur, Andhra Pradesh, India.
INTRODUCTION
Since ancient times, about 80 % of individuals
use traditional medicine, which has chemical
compounds derived from medicinal plants
1, 2
.
Several hundred plant species and herbs in the
form of whole plant, crude extract or purification,
purified constituents are used in indigenous
system of medicines and are of great
importance to the health of individual and
communities, which have ultimately evaluated
into the modern therapeutic science. Medicinal
plants are important source of life saving drugs
for majority of the world population
3, 4
.
Research Article
ABSTRACT
Phytochemical constituents are non-nutritive plant chemicals that have preventive and curative
properties of disease. The use of plants and phytochemicals, both with known biological properties,
can be of great significance in therapeutic treatment. The present study includes phytochemical
screening and quantification of secondary metabolites and their biological activities of root extract
of Syzygium samarangense. Phytochemical screening of the plant root extracts with ethyl acetate,
methanol and water showed the presence of flavonoids, terpenoids and phenolic compounds. In
aqueous extracts, Terpenoids with highest quantity (81.923 micrograms per gram extract) were
estimated, whereas flavonoids are present only in methanolic extract and with an estimated quantity
of 33.687 µg /per gram extract. Anti oxidant, anti inflammatory, and anti diabetic activity of these
root extracts reveal that methanoilc extracts with flavonoids showed the high antioxidant, anti
inflammatory and antimicrobial activity than the aqueous and ethyl acetate extract. Highest
antidiabetic activity was observed in aqueous extract. Antimicrobial activity study reveals that gram
positive bacterial strains are more sensitive than the gram negative bacteria to all the three types of
root extracts. The present study reveals that the root extract of Syzygium samarangense is a potential
source for phytochemicals for traditional use as therapeutics.
Keywords: Syzygium samarangense (wax jambu), flavonoids, terpenoids.
IJRPC 2015, 5(4), 753-763 Madhavi et al.
ISSN: 2231
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754
Fig. 1: Syzygium samarangense (wax jambu) plant and fruit
Syzygium samarangense (common name - wax
jambu) is a plant species in the family Myrtaceae
which is widely cultivated in the tropics
5-13
. There
are lot of traditional claims has been reported of
leaves, root, bark, fruits of the plant
14-17
. Various
Pharmacological Activities like Antidiarrhoeal
Activity,
Anticholinesterase
Activity,
Immunopharmacological
Activity,
Cytotoxic
Activity, Anti hyperglycaemic activity, Analgesic
and Anti- Inflammatory activity are reported with
various parts of the plant
18-25
. And also
Investigators
have
found
their
principal
constituent to be tannins, Quercetin glycosides,
monoterpenes secondary metabolites those
involved in pharmacological properties
26, 27
.
Traditionally the root bark decoction of the
Syzygium samarangense is used in dysentery
and amenorrhea and also used as abortifacient.
Root is used as diuretic and is given to alleviate
edema. Malayans use powdered dried root
preparations for itching. As plant root has
significant therapeutic uses, this study is aimed
to screen the phytochemicals of the root and
study of its biological activities.
The literature review proves that the plant is rich
with many medicinal and bioactive compounds
like flavanoids, phenolic compounds, glycosides,
terpinoids etc. various studies are reported
these compounds in various areal parts of the
plant. K. M. Moneruzzaman et al
14
, reported the
flavonoids (914.1 mg/100g) and phenolic
(326.67 mg GAE/100g) content from the
Syzygium samarangense plant where in our
study root is reported as 3mg/100grams. Wu
YZ
et
al
24
,
investigated
the
chemical
constituents from the branches and leaves with
95% ethanol, then partitioned with petroleum
ether, chloroform and ethyl acetate. M.O.
Edema
et
al
36
,
studied
on
Syzygium
samarangense juice extracts and evaluated high
amounts of saponins content (4.77%). Mario J.
Simirgiotis et al
37
, reported the six quercetin
glycosides on the methanolic extracts of the pulp
and
seeds
of
the
fruits
of
Syzygium
samarangense Merr. & Perry (Blume). Vasanthi
et al
38
, reported the total phenolic content
(162.58±0.51 μg/mg), total flavonoid contents
(310μg/mg) of Syzygium samarangense Fruit
extract (SSFE) where the study with roots are
reported 46.944 µg and 23.056 µg per gram in
methanolic and water extract respectively. G. R
Nair et al
39
, reported the two flavonol
glycosides. Ghayur MN et al [40], reported four
flavonoids isolated from the hexane extract.
Evangeline C. Amor et al
18
, isolated four rare
C-methylated flavonoids with a chalcone and a
flavanone the compounds from the hexane
extract of the leaves. Dennis D. Raga et al
41
,
isolated the Cycloartenyl stearate ( 1a), lupenyl
stearate (1b), sitosteryl stearate (1c), and 24-
methylenecycloartanyl stearate (1d) (sample 1)
from the air-dried leaves. Samy MN et al
42
,
isolated three new compounds (one new
cyanogenic glucoside) from a MeOH extract of
the leaves. Rachana Srivastava et al
43
, isolated
the new triterpene, methyl 3epibetulinate in its
native form and 4′,6′dihydroxy2′ methoxy3′,
5′dimethyl chalcone along with ursolic acid,
jacoumaric acid and arjunolic acid have been
isolated from the aerial parts. However the study
with root extracts reported that root reported with
high terpenoid content which is not reported
earlier. And it is also reported that root is
another alternate source for flavonoid and
phenoilc groups. Anti oxidant, anti inflammatory,
and anti diabetic activity of these three root
extracts conducted in order to study the
biological activity of the plant. Anti oxidant
activity of the extract was conducted by DPPH
scavenging
activity.
Results
reveal
that
methanolic extracts which consist of flavonoids
has shown the high antioxidant activity
IJRPC 2015, 5(4), 753-763 Madhavi et al.
ISSN: 2231
2781
755
(88.021%) (table-3 and graph A) than the
aqueous and ethyl acetate extracts at 40 µg/ml
extract concentration. Similar studies are
reported by the Fonseca A et al
44
, investigated
the antioxidant activity (0.9 μMol TROLOX®/mg)
of the fruit. Vasanthi et al
38
, reported that IC50
value of antioxidant activity of the fruit extract
found to be 140 μg/ml in standard (Lascorbic
acid), 280 μg/ml in SSFE by DPPH and
ABTS.+ scavenging activity found to be the IC50
value 175 μg/ml and 250 μg/ml respectively.
No one has reported with root extracts and
found that similar antioxidant activity as fruit.
Antidiabetic activity (table-4 and graph B) of the
root extracts was studied by In vitro
α
- amylase
inhibition activity by Spectrophotometric method.
Results showed that dose dependent inhibition
was observed with all the samples where as
aqueous extract (92.626%) exhibited the highest
activity than remaining two extracts. No
antidiabetic activity studies are reported with S.
Samarangense. Anti inflammatory activity (table-
5) of the root also studied with all three extracts
and among them methanolic extracts (84.552%)
showed the highest anti inflammatory activity. In
literature Shabnam Mollika et al
45
, reported
evaluated the moderate effect anti-inflammatory
activity of the methanolic extract of leaves in
mice. Dennis D. Raga et al
41
, reported that the
leaves exhibited potent analgesic and anti-
inflammatory activities at effective doses of 6.25
mg/kg body weight and 12.5 mg/kg body weight,
respectively. This reveals theat no one has
reported the anti inflammatory activity with root
and potential activity was reported in our study.
Antimicrobial activity result (table-6 and figure-3)
reveals that Gram-positive bacteria were more
sensitive than Gram-negative ones towards the
plant extracts studied. Bacillus subtilis and
Staphylococcus aureus strains are shown more
inhibition zone than gram negative Escherichia
coli, Pseudomonas aeruginosa, Salmonella
typhi. Previous studies are also reported the
antimicrobial activity of the different areal plant
extracts. S.
Adeola
Adesegun
et
al
46
,
evaluated the antimicrobial effect of the volatile
oil from the leaf of Syzygium samarangense on
Escherichia coli. Abd Aziz et al
47
, evaluated
antimicrobial properties of ethanolic extracts of
the leaves of minimum inhibitory concentration
(MIC) value was determined to be 20 mg/mL. K.
Venkata Ratnam et al
48
, evaluated the
antimicrobial properties of fruits, against certain
bacterial and fungal strains with petroleum ether
and methanol extracts found to be effective on
both Gram positive and Gram negative bacteria.
M.O. Edema et al
36
, reported that juice extracts
have significant (P<0.05) antimicrobial activities
against Escherichia coli, Salmonella typhi and
Candida albicans. Consolacion Y. Ragasa et
al
49
, elucidated the dichloromethane extract of
the leaves exhibited moderate antifungal activity
against C. albicans, low activity against T.
mentagrophytes and low antibacterial activity
against E. coli, S. aureus and P. aeruginosa. It
was inactive against B. subtilis and A. niger. In
comparison with the literature studies root
extracts of S. samarangense also have
significant antimicrobial active compounds with
low minimum inhibition concentration. Studies
proved that compounds in plant extracts have
potential activity against gram positive bacteria
than gram negative.
MATERIAL AND METHODS
Chemicals: The solvents used for root
extraction are Methanol and Ethyl Acetate. The
reagents used for phytochemical screening and
estimation were of laboratory reagent grade and
were purchased for Merck chemicals private
limited, Mumbai, Fisher scientific, Mumbai and
SD fine chemicals Mumbai. Distilled water has
been used for aqueous extraction. Alpha
amylase enzyme was purchased from Sigma
Aldrich chemicals, Ofloxacin drug.
Apparatus:
Denver
electronic
balance,
TECHCOMP
–
UV 2301 Double Bean UV
Visible Spectrophotometer with HITACHI 2.2
software,
Tech-comp
UV
visible
spectrophotometer,
soxhlet
extraction
apparatus, heating mantle, incubator, autoclave.
Sample collection: Root material of Syzygium
samarangense plants were collected from farms
in various places of East Godavari district,
Andhra Pradesh, India. The roots are separated
and allowed to shade dry. The root sample was
ground and powdered for solvent extraction.
Solvent
extraction:
The
phytochemicals
present in the roots of the collected plants were
isolated using different solvents like ethyl
acetate, methanol and water in a series of
extraction method from low polarity to high
polarity using soxhlet extraction method.
Microbial test strains for antibiotic activity
The bacterial strains used for screening of
antimicrobial activity are Salmonella typhi,
Escherichia coli, Pseudomonas aeruginosa,
Bacillus subtilis.
IJRPC 2015, 5(4), 753-763 Madhavi et al.
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Phytochemical analysis of the root extracts was
performed by following standard methods
Preliminary phytochemical screening
1. Test for steroids
Salkowski Test
Few drops of concentrated sulphuric acid are
added to the plant extract, shaken and on
standing; lower layer turns red in colour.
Liebermann Burchard’s Test
To the extract, few drops of acetic anhydride
was added and mixed well. 1 ml of
concentrated sulphuric acid is added from the
sides of test tube, a reddish brown ring is
formed at the junction of two layers.
2. Tests for triterpenoids
Salkowski Test
Few drops of concentrated sulphuric acid are
added to the extract, shaken and on
standing, lower part turns golden yellow
colour.
Lieberman Burchard’s Test
To the extract, few drops of acetic anhydride
was added and mixed well. 1 ml of
concentrated sulphuric acid is added from the
sides of test tube, a red ring indicates
triterpenes.
Ischugajiu Test
Excess of acetyl chloride and pinch of zinc
chloride are added to the extract solution,
kept aside for reaction to subside and
warmed on water bath, cosin red colour is
produced.
Brickorn and Brinar Test
To the extract, few drops of chlorosulfonic
acid in glacial acetic acid (7:3) are added, red
colour is produced.
3. Test for Saponins
Foam Test
Small amount of extract is shaken with little
quantity of water; the foam produced persists
for 10 minutes. It confirms the presence of
saponins.
Haemolysis Test
To 2ml of 1.8% Sodium chloride solution in
two test tubes, 2ml distilled water is added to
one and 2ml of 1% extract to the other, 5
drops of blood is added to each tube and
gently mixed with the contents. Haemolysis
observed under the microscope in the tube
containing the extract indicates the presence
of saponins.
4. Test for Steroidal Saponin
The extract is hydrolysed with sulphuric acid and
extracted with chloroform. The chloroform layer
is tested for steroids.
5. Tests for Triterpenoidal Saponin
The extract is hydrolysed with sulphuric acid and
extracted with chloroform. The chloroform layer
is tested for triterpenoids.
6. Tests for Alkaloids
Mayer’s Test
The acid layer when mixed with Mayer’s
reagent (Potassium mercuric iodide solution)
gives creamy white precipitate.
Dragendroff’s Test
The acid layer with a few drops of
Dragendroff’s
reagent (Potassium bismuth
iodide) gives reddish brown precipitate.
Wagner’s Test
The acid layer when mixed with few drops of
Wagner’s reagent (solution of iodide in
potassium iodide) gives brown to red
precipitate.
Hager’s Test
The acid layer when mixed with few drops of
Hager’s reagent (Saturated solution of pricric
acid) gives yellow coloured precipitate.
7. Tests for Carbohydrates
Fehling
’s Test
The extract when heated with Fehling’s A
and B solutions gives an orange red
precipitate showing the presence of reducing
sugar.
Molisch
’s Test
The extract is treated with Molisch’s reagent
and concentrated sulphuric acid along the
sides of the test tube, a reddish violet ring
shows the presence of carbohydrate.
Benedict’s test
The extract on heating with
Benedict’s
reagent, brown precipitate indicates the
presence of sugar.
Barfoed’s Test
Barfoed’s reagent is added and boiled on
water
bath
for
few
minutes;
reddish
precipitate is observed for the presence of
carbohydrate.
8. Test for Flavonoids
Shinoda Test
The extract solution with a few fragments of
magnesium
ribbon
and
concentrated
hydrochloric acid produced magenta colour
after few minutes.
IJRPC 2015, 5(4), 753-763 Madhavi et al.
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757
Ferric chloride test
Alcoholic solution of extract reacts with
freshly prepared ferric chloride solution and
given blackfish green color.
Lead Acetate Test
Alcoholic solution of extract reacts with 10%
lead acetate solution and given yellow
precipitate.
9. Test for Glycosides
Anthraquinone test
Drug is powdered and extracted with either
ammonia or caustic soda. The aqueous layer
shows pink color
Keller-killiani test
This is for cardiac glycosides. The extract
and 0.4 glacial acetic acid are mixed with
ferrous chloride and 0.5 mi of concentrated
sulphuric acid. The acetic acid layer shows
blue color.
10. Test for Phenolic Compounds
Ferric chloride test
Treat the extract with ferric chloride solution
then blue color appears if hydrolysable
tannins are present and green color appears
if condensed tannins are present.
Gelatin test
To the test solution add 1% gelatin solution
containing 10% NaCl, and then precipitate is
formed.
Test for chlorogenic acid
Treat the test solution with aqueous ammonia
and expose to air gradually, green color is
developed.
Quantitative
analysis
of
Phenoilc
Compounds
The total phenolic content in different solvent
extracts was determined with the Folin-
Ciocalteu’s reagent (FCR). In the procedure, 1ml
of extract was mixed with 0.4 ml FCR (diluted
1:10 v/v). After 5 min 4 ml of sodium carbonate
solution was added. The final volume of the
tubes were made up to 10 ml with distilled water
and allowed to stand for 90 min at room
temperature. Absorbance of sample was
measured against the blank at 765nm using a
spectrophotometer. A calibration curve was
constructed using gallic acid solution as
standard and total phenolic content of the
extract was expressed in terms of milligrams of
gallic acid per gram of dry weight.
Determination of total flavonoid content
Total flavonoid content was determined using
aluminium chloride (AlCl
3
) according to a known
method using quercetin as a standard. The plant
extract (1 ml) was added to 3 ml distilled water
followed by 5% NaNO
2
(0. 3ml). After 5 min at
25°C, AlCl
3
(0.3 ml, 10%) was added. After
further 5 min, the reaction mixture was treated
with 2.0 ml of 1 M NaOH. Finally, the reaction
mixture was diluted to 10ml with water and the
absorbance was measured at 510 nm. A
calibration
curve
was
constructed
using
quercetin solutions as standard and total
phenolic content of the extract was expressed in
terms of milligrams of quercetin per gram of dry
weight.
Quantitative estimation of terpenoids
To 1ml of plant extract 2 ml of chloroform was
mixed in extract of plant sample and 3 ml of
sulphuric acid were added in sample extract.
Reddish brown color was obtained in the test
tube. Final volume in the test tube was made up
to 10ml with water. The absorbance of the
formed color was measured at 538nm against
reagent blank prepared similarly without plant
extract. Linalool
was
used
as
standard
terpenoid.
Measurement of Antioxidant Activity using
DPPH method
The antioxidant activity of the different root
extracts was determined on the basis of their
scavenging activity of the stable 1, 1- dipheny1-
2-picryl hydrazyl (DPPH) free radical. DPPH is a
stable free radical containing an odd electron in
its structure and usually utilized for detection of
the radical scavenging activity in chemical
analysis. lml of each solution of different
concentrations (1-500g/m1) of the extracts was
added to 3 ml of 0.004% ethanolic DPPH free
radical
solution.
After
30
minutes
the
absorbance of the preparations were taken at
517 nm by a UV spectrophotometer which was
compared with the corresponding absorbance of
standard ascorbic acid concentrations (1-
500µg/m1).The method described by Hatano et
al was used to measure the absorbance with
some modifications. Then the % inhibition was
calculated by the following equation:
IJRPC 2015, 5(4), 753-763 Madhavi et al.
ISSN: 2231
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Antidiabetic activity
In vitro study α
- amylase inhibition activity
by Spectrophotometric method
1ml of alpha amylase and 1 ml of plant extract in
a test tube and incubated at 37
◦
C for 10 min.
After pre-incubation, 1ml of 1% (v/v) starch
solution was added to each tube and incubated
at 37◦C for 15min. The reaction was terminated
with 2 mL DNSA reagent, placed in boiling water
bath for 5min, cooled to room temperature,
diluted, and the absorbance measured at 546
nm. The control reaction representing 100%
enzyme activity did not contain any plant extract.
To eliminate the absorbance produced by plant
extract, appropriate extract controls were also
included.
% inhibition of alpha amylase by each plant extract can be calculated using the formula:
Anti inflammatory Activity by Albumin
denaturation Assay
A solution of 0.2% W/V of BSA was prepared in
Tris buffer saline and PH was adjusted to 6.8
using glacial acetic acid. Stock solutions of plant
extract were prepared by using methanol as a
solvent. From these stock solutions 6 different
concentrations of 10-500µg/ml were prepared by
using methanol as a solvent. 50µl of each
extract was transferred to Eppendorf tubes using
1ml micro pipette. 5ml of 0.2% W/V BSA was
added to all the above Eppendorf tubes. The
control consists of 5ml 0.2% W/V BSA solution
with 50 µl methanol. The test tubes were heated
at 72° C for 5 minutes and then cooled for 10
minutes. The absorbance of these solutions was
determined by using UV/Vis Double beam
spectrophotometer
(Elico
SL-196)
at
a
wavelength of 660nm. The % inhibition of
precipitation (denaturation of the protein) was
determined on a % basis relative to the control
using the following formula.
Antimicrobial Activity
Root extracts of Syzygium samarangense were
tested by agar well-diffusion method to
determine the antimicrobial activity. Nutrient
agar (NA) plates were seed inoculated. Wells
(10mm diameter and about 2 cm a part) were
made in each of these plates using sterile cork
borer. Stock solution of each plant extract was
prepared at a different concentrations 1000,
500, 200, 100, 50,10µg/ ml in different plant
extracts viz. Methanol, ethyl acetate, water.
About 100 µl of different concentrations of plant
solvent extracts were added with sterile syringe
into the wells and allowed to diffuse at room
temperature for 2hrs. Control experiments
comprising inoculum without plant extract were
set up. The plates were incubated at 37°C for
18-24 h. The diameter of the inhibition zone
(mm) was measured and the activity index was
also calculated. Triplicates were maintained and
the experiment was repeated thrice, for each
replicates the readings were taken in three
different fixed directions and the average values
were recorded. Oflaxacin drug was used as
standard antibacterial agent and compared with
the standard results.
RESULT AND DISCUSSION
Secondary metabolites present in the plants are
responsible for the biological activities such as
hypoglycaemic,
antidiabetic,
antioxidant,
antimicrobial, antiinflammatory, anticarcinogenic,
antimalarial, anticholinergic, antileprosy etc. The
preliminary screening of phytochemicals and
evaluation of bioactive may lead to medicinal
plant drug discovery and development of
phytomedicine. In the present study the root of
Syzygium samarangense (wax Jambu) was
screened
for
determination
of
it’s
phytochemicals
in
three
different
solvent
systems. Among the three, aqueous extracts
were proved to contain more number of
compounds than other two solvents extracts. In
aqueous
extract,
alkaloids,
carbohydrates,
saponins, tannins, roteins and aminoacids,
terpenoids, phenolic compounds were identified.
Tannins,
Flavonoids,
terpenoids,
Phenolic
Compounds were indentified in the methanolic
IJRPC 2015, 5(4), 753-763 Madhavi et al.
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2781
759
extract. In Ethyl acetate extract only Terpenoids
are identified. In continuation quantitative
analysis of the extracts has conducted to
estimate terpenoids, phenolic compounds and
flavonoids (table 1 and fig 2).
Terpenoids were detected with highest quantity
(81.923 micrograms per gram extract) in
aqueous extracts than other phytochemicals.
Terpenoids were detected with 48.461µg
quantity in methanolic extract and 35.385 µg in
ethyl acetate extract. Whereas flavonoids are
only present in methanolic extract and amount of
flavanoid quantified was 33.687 µg /per gram
extract. Similarly phenolic compounds were
reported in methanolic and water extracts with
estimates as 30.156µg/g and 23.056 µg per
gram extract respectively. Our results were in
agreement with findings of the medicinal value
of plants lies in some chemical substances that
have definite physiological functions in the
human body. Different phytochemicals have
been found to possess a wide range of
medicinal properties, which may help in
protection against various diseases.
CONCLUSION
In the present investigation, primary and
secondary metabolites of the root were
qualitatively and quantitatively analyzed then the
biological
activity
(anti
oxidant,
anti
inflammatory, and anti diabetic) was studied.
Further evaluation of phytochemicals and their
activity is needed for knowing the nutritional
potential as well as helpful in manufacturing new
drugs.
ACKNOWLEDGEMENTS
The author is thankful to Head, Dept. of Botany
& Microbiology, ANU for providing the facilities
and also thankful to management, Hindu
College
of
Pharmaceutical
sciences
for
permitting to do research analysis.
Table 1: Phytochemical screening of root extracts of Syzygium samarangense
S. No.
Name of the tests
Ethyl acetate extract
Methanolic extract
Water extract
1
Alkaloids
-ve
-ve
+ve
2
Carbohydrates
-ve
-ve
+ve
3
Glycosides
-ve
-ve
-ve
4
Saponins
-ve
-ve
+ve
5
Tannins
-ve
+ve
+ve
6
Proteins & Aminoacids
-ve
+ve
+ve
7
Flavanoids
-ve
+ve
-ve
8
Terpenoids
+ve
+ve
+ve
9
Phenolic Compounds
-ve
+ve
+ve
Flavanoid test Terpenoid test
Fig. 2: images of phytochemical screening of root extract of Syzygium samarangense (flavonoids,
terpenoids tests)
IJRPC 2015, 5(4), 753-763 Madhavi et al.
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Table 2: Quantitative analysis of root extracts of S. samarangense
Phytochemical
Extract
Amount found µg / g of extract
Terpenoids
Ethyl Acetate
35.385
Methanolic
48.461
Aqueous
81.923
Flavonoids
Methanolic
33.687
Phenolic Compounds
Methanolic
30.156
Water
23.056
Table 3: Antioxidant activity (% DPPH scavenging activity) of S. samarangense root extracts
S. No.
Concentration in µg/ml
Ascarbic Acid
EtOAc Extract
MeOH
Extract
Water
Extract
1
2
13.262
2.246
6.417
3.422
2
4
22.460
4.492
18.930
8.449
3
6
37.005
13.155
33.155
18.930
4
8
56.684
21.283
49.519
28.449
5
10
74.011
33.155
76.898
45.134
6
20
89.519
52.620
78.930
63.422
7
40
90.695
60.535
88.021
75.508
% DPPH Activity
Graph A: Comparative graph of DPPH assay of S. samarangense root extracts
Table 4:
α
- amylase inhibition activity of S. samarangense root extracts
S. No.
Concentration of Sample
EtOAc Extract
MeOH Extract
Water Extract
% of α
-Amylase inhibition
1
5µg/ml
7.709
13.408
17.207
2
10µg/ml
9.385
20.223
28.603
3
15µg/ml
11.173
26.033
41.341
4
20µg/ml
21.341
34.637
52.849
5
25µg/ml
30.168
51.285
59.665
6
50µg/ml
38.436
75.307
76.201
7
100µg/ml
51.285
79.218
86.480
8
200µg/ml
55.866
88.715
92.626
0
0.2
0.4
0.6
0.8
1
0
20
40
60
Abs
o
rba
nce
Concentration in µg/ml
Conc Vs Abs
Ascarbic
Acid
EtOAc
Extract
MeOH
Extract
Water
Extract
0
20
40
60
80
100
0
10
20
30
40
50
%DP
P
H
inh
ibi
tio
n
Concentration in µg/ml
Conc Vs % DPPH Inhibition
Ascarb
ic Acid
EtOAc
Extract
IJRPC 2015, 5(4), 753-763 Madhavi et al.
ISSN: 2231
2781
761
Graph B: Comparative graph of α
- amylase inhibition activity of S. samarangense root extracts
Table 5: Anti inflammatory activity of S. samarangense by Albumin denaturation assay
S. No.
Concentration of
Sample
EtOAc Extract
MeOH Extract
Water Extract
% Albumin denaturation
1
10µg/ml
5.241
27.724
15.724
2
50µg/ml
13.793
35.034
19.724
3
100µg/ml
23.724
44.000
29.379
4
200µg/ml
36.414
55.448
50.069
5
400µg/ml
54.621
68.828
60.138
6
500µg/ml
60.827
84.552
70.759
Table 6: Anti microbial activity of S. samarangense root extracts
S. No.
Test organism
Size of zones (in mm)
methanolic
ethyl acetate
aqueous
Standard
1000 µg/ml
1000 µg/ml
1000 µg/ml
500 µg/ml
1
Salmonella typhi
12.7
….
5.6
19.5
2
Escherichia coli
19.2
11.2
17.5
28.7
3
Pseudomonas aeruginosa
15.3
8.7
11.6
21.2
4
Bacillus subtilis
19.5
14.3
16.5
31.5
1) Salmonella typhi 2) Escherichia coli 3) Pseudomonas aeruginosa 4) Bacillus subtilis
Fig. 3: Antimicrobial activity results of S. samarangense root extracts
0
20
40
60
80
100
0
2
4
6
8
10
%
α
-
a
m
y
la
se
inh
ibi
tio
n
Concentration in µg/ml
α- amylase inhibition activity
EtOAc Extract
MeOH Extract
Water Extract
IJRPC 2015, 5(4), 753-763 Madhavi et al.
ISSN: 2231
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REFERENCES
1. "Syzygium samarangense (Blume) Merr.
&
L.M.Perry".
World
Checklist
of
Selected Plant Families (WCSP). Royal
Botanic Gardens, Kew. Retrieved 14
Mar 2016
–
via The Plant List.
2. Julia F. Morton (1987). "Java apple".
Fruits of Warm Climates. Miami, FL:
Florida Flair Books. pp. 381
–
382.
3. " Syzygium samarangense (Blume) Merr.
& L.M.Perry". Germplasm Resources
Information Network (GRIN). Retrieved
14 Mar 2016.
4.
Peter, Tina; Padmavathi, D (28 October
2011). "Syzygium Samarangense: A
Review On Morphology, Phytochemistry
& Pharmacological Aspects" (PDF).
Asian Journal of Biochemical and
Pharmaceutical Research1 (4): 155.
Retrieved 1 June 2015.
5. Whitman, William F.Five Decades with
Tropical Fruit, a Personal Journey.
Stuart, Florida: Quisqualis Books in
cooperation
with
Fairchild
Tropical
Garden. 2001. Print
6. "Syzygium samarangense, Syzygium
javanicum,
Eugenia
javanica".
toptropicals.com. N.d. Web. 9 Dec.
2014.
7. " Syzygium
samarangense."
worldagroforestrycenter.org.
Web.
1
Mar. 2015.
8. Whitman, William F. CSIR. 1976. The
Wealth of India: Raw materials. Vol X
Sp-W. CSIR.
9. Jensen M. 1995. Trees commonly
cultivated in South-East Asia: An
illustrated field guide. RAP publication:
1995/38.Bangkok, Thailand. 229pp.
10. Martin FW, Campbell CW & Ruberte
RM. 1987. Perennial edible fruits of
tropics: an inventory. US Department of
Agriculture, Agriculture Handbook No.
642. 252 pp.
11. Okuda, TT, Yoshida, Hatamo, T, Yazaki,
K, & Ashida, M. 1982. Ellagitannins of
the Casuarinaceae, Stachyuraceae and
Myrtaceae.
Phytochemistry.
21(12):
2871-2874.
12. Panggabean
G.1992.
Syzygium
aqueum
(Burm.f.)
Alst.,
Syzygium
malaccense (L.) M. & P, and Syzygium
samarangense (Blume) M. & P. In
Coronel, R.E., et al. (Eds.): PROSEA.
No. 2: Edible fruits and nuts. Prosea
Foundation, Bogor, Indonesia. pp. 292-
294.
13. Walter A, Sam C. 2002. Fruits of
Oceania. ACIAR Monograph No. 85.
Canberra.329 pp.
14. A.M. Al-Saif, H. Sharif, M.T. Rosna &
K.M. Moneruzzaman., African Journal of
AgriculturalResearch.,
2011,
6(15),
3623.
15. M.N. Ghayur, A.H. Gilani, A. Khan, E.C.
Amor, I.M. Villaseñor & M.I. Choudhary.,
Phytother Res., 2006, 20(1), 49.
16. K.A. Reynertson, M.J. Basile & E.J.
Kennelly., Ethnobotany Research &
Applications., 2005,3, 25.
17. A.W. Whistler, W. Arthur., Species
Profiles
for
Pacific
Island.,
Agroforestrywww.traditionaltree.org
[online], 2010.
18. C.A. Evangeline, M.V. Irene, A.N.
Sarfraz, Sabir & M.C. Iqbal., Philippine
Journal of Science., 2005, 134 (2), 105.
19. Y.C. Kuo, L.M. Yang & L.C. Lin., Planta
Med., 2004, 70(12), 1237.
20. Amor, E. Villaseñor, Irene, Antemano,
Rowena et al., Pharmaceutical Biology
(Formerly
International
Journal
of
Pharmacognosy)., 2007, 45., 777.
21. M.H.
Resurreccion-Magno,
I.M.
Villaseñor, N. Harada & K. Monde.,
Phytother Res., 2005, 19(3), 246.
22. K.V. Ratnam & V.R. Raju., Advances in
Biological Research., 2008, 1-2, 17.
23. D.D. Raga, C.L. Cheng, K.C. Lee, W.Z.
Olaziman, V.J. De Guzman, C.C. Shen
et al., Z Naturforsch., 2011, 66 (5-6),
235.
24. T. Soubir & Sheng Wu Gong Cheng Xue
Bao., 2007, 23(2), 257.
25. A.R. Kurt, Y. Hui, J. Bei, J.B. Margaret &
J.K. Edward., FoodChemistry., 2008,
109, 883.
26. C.A. Evangeline, M.V. Irene, Y. Amsha
& C. Iqbal., Prolyl Endopeptidase
Inhibitors from Syzygium samarangense
(Blume)., Merr. & L. M. Perry. Z.
Naturforsch, 2004, 59c, 86.
27. A.G. Nair, S. Krishnan, C. Ravikrishna
& K.P. Madhusudanan., Fitoterapia.,
1999, 70, 148.
28. Dey PM and Harborne J B. Methods in
Plant Biochemistry: Academic Press;
London, 1987.
29. Evans WC. Pharmacognosy,13th Ed,
Balliere
–
Tindall; London, 1989.
IJRPC 2015, 5(4), 753-763 Madhavi et al.
ISSN: 2231
2781
763
30. Evans
WC.
Trease
and
Evans
Pharmacognosy, 14th Edition, Bailiere
Tindall W.B. Sauders company ltd;
London, 1996, 224
–
228, 293
–
309,
542
–
575.
31. Parekh J and Chanda SV. In vitro
antimicrobial activity and phytochemical
analysis of some Indian medicinal
plants. Turk J Biol. 2007;31:53-58.
32. Katasani
Damodar.
Phytochemical
screening, quantitative estimation of
total
phenolic,
flavanoids
and
antimicrobial
evaluation
of
trachyspermum ammi. J Atoms and
Molecules. 2011;1(1):1
–
8.
33. Hatano, T., H. Kagawa, T. Yasuhara
and T.
Okuda, 1988.
Two new
flavonoids and other constituents in
licorice root: their relative astringency
and radical scavengingeffects. Chem.
Pharm. Bull., 36: 1090-2097.
34. Varun Kumar Prabhakar., International
Journal of Scientific and Research
Publications, Volume 3, Issue 8, August
2013
35. Williams LAD, Connar AO, Latore L et
al. The in vitro anti-denaturation effects
induced
by
natural
products
and
nonsteroidal compounds in heat treated
(immunogenic) bovine serum albumin is
proposed as a screening assay for the
detection
of
anti-inflammatory
compounds, without the use of animals,
in the early stages of the drug discovery
process. West Indian Med J, 57: 327
–
331, (2008)
36. M.O.
Edema
et
al,
Comparative
evaluation of bioactive compounds in
hibiscus
sabdariffa
and
syzygium
samarangense juice extracts, African
Crop Science Journal, Vol. 20, No. 3,
pp. 179
–
187.
37. Mario J. Simirgiotis et al, Cytotoxic
chalcones and antioxidants from the
fruits of a Syzygium samarangense
(Wax Jambu), Food Chem. 2008 Mar
15; 107(2): 813–
819.
38. Vasanthi et al, In vitro antioxidant
activity of Syzygium samarangense
merr. et perry. Fruit extract, Journal of
Pharmacy Research;2
012, Vol. 5 Issue
6, p 3426.
39. A. G. R Nair et al, New and rare flavonol
glycosides from leaves of Syzygium
samarangense,
in
FITOTERAPIA
70(2):148-151 · APRIL 1999.
40. Ghayur MN et al, Presence of calcium
antagonist activity explains the use of
Syzygium samarangense in diarrhea,
Phytother Res. 2006 Jan;20(1):4952.
41. Dennis D. Raga et al, Bioactivities of
Triterpenes and a Sterol from Syzygium
samarangense, Z. Naturforsch. 66 c,
235
–
244 (2011).
42. Samy MN et al, Taxiphyllin 6'Ogallate,
actinidioionoside
6'Ogallate
and
myricetrin 2″Osulfate from the leaves of
Syzygium samarangense and their
biological activities, Chem Pharm Bull
(Tokyo). 2014;62(10):10138.
43. Rachana
Srivastava
et
al,
Plant
chemistry Triterpenoids and chalcone
from
Syzygium
samarangense,
Phytochemistry, Volume 38, Issue 3,
February 1995, Pages 687
–
689.
44. Fonseca A et al, Antioxidant activity of
ethanol extract of the fruit of the species
Syzygium samarangense (WAX APPLE)
. J Pharm Pharmacogn Res (2014)
2(Suppl. 1):S59
45. Shabnam Mollika et al, Evaluation of
Analgesic, Anti-Inflammatory and CNS
Activities of the Methanolic Extract of
Syzygium samarangense Leave, Global
Journal of Pharmacology 8 (1): 39-46,
2014.
46. S. Adeola Adesegun et al, Essential Oil
of Syzygium samarangense; A Potent
Antimicrobial and Inhibitor of Partially
Purified and Characterized Extracellular
Protease of Escherichia coli 25922 ,
British Journal of Pharmacology and
Toxicology 4(6): 215-221, 2013.
47. Abd Aziz et al, Screening of selected
Malaysian plants against several food
bore pathogen bacteria, International
Food Research Journal 18(3): 1195-
1201 (2011).
48. K. Venkata Ratnam et al, In vitro
Antimicrobial Screening of the Fruit
Extracts of Two Syzygium Species
(Myrtaceae), Advances in Biological
Research 2 (1-2): 17-20, 2008.
49. Consolacion Y. Ragasa et al, Chemical
constituents
of
Syzygium
samarangense, Der Pharma Chemica,
2014, 6(3):256-260.
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