See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/236246954

Bioactive compounds in Sesbania sesban flower and its Antioxidant and

Antimicrobial activity
Article in Journal of Pharmacy Research · December 2012

CITATIONS READS
2 638
3 authors:

Saravanakumar Marappan Kathiresh Manickam

Rasi seeds 4 PUBLICATIONS 5 CITATIONS

2 PUBLICATIONS 5 CITATIONS
SEE PROFILE
SEE PROFILE
P.Suganya Devi

NGM College
15 PUBLICATIONS 220 CITATIONS

SEE PROFILE

All content following this page was uploaded by Kathiresh Manickam on 30 May 2014.

The user has requested enhancement of the downloaded file.

 M. Kathiresh et al. / Journal of Pharmacy Research 2012,5(1),390-293
Research Article Available online through
ISSN: 0974-6943 www.jpronline.info
Bioactive compounds in Sesbania sesban flower
and its Antioxidant and Antimicrobial activity
M. Kathiresh , P.Suganya Devi and M.Saravanakumar *
P.G & Research Department of Biotechnology,Dr. Mahalingam Center for research and development, N.G.M.College, pollachi,Tamil Nadu, India
Received on:20-09-2011; Revised on: 15-10-2011; Accepted on:10-12-2011

ABSTRACT
In plants, there are number of bioactive compounds present, among them anthocyanins are water – soluble natural pigments found in flowers and fruits .
The pigments may appear in different colours such as red, purple or blue. They belong to a parent class of molecules called flavonoids synthesized
through the phenylpropanoid pathway. Anthocyanins occur in all tissues of higher plants, including leaves, stems, roots, flowers and fruits.
Anthocyanins are derivatives of anthocyanidins which include pendant sugars. This work was an attempt to extract anthocyanin from Sesbania sesban
flower petals. Anthocyanins from coloured petals of Sesbania sesban flower were extracted by using solvents such as methanol and acidified methanol.
The studies were carried out from the crude extracts to obtained anthocyanins, total phenol and total flavonoids from flower petals. From the extracts of
Sesbania sesban flower petals the antioxidant and antimicrobial properties of anthocyanins were analysed. These merits can put the plant in a wider and
more significant perspective with respect to national and global interest in medicinal uses and food industries.

Key words: Sesbania sesban, Anthocyanin, Antioxidant activity, Antimicrobial activity.

INTRODUCTION
Anthocyanins (also anthocyans), the secondary metabolites produced by the linoleic acid peroxidation. The realization that many infectious pathogenic
plants are water- soluble pigments which is responsible for colours like organisms are fast developing resistance (s) against the prevailing drugs
purple , blue, pink ,red in the tissues of higher plants, including leaves, stems, has necessitated a search for new sources of antibacterial and antioxidant
roots, flowers, and fruits. Anthocyanins are derivatives of anthocyanidins, they com-pounds. They are expected to synthesize a variety of secondary
are odorless and nearly flavorless, contributing to taste as a moderately metabolites capable of providing them.
astringent sensation. In addition to their role as light-attenuators, anthocya-
nins also act as powerful antioxidants. In recent years, there has been increas- MATERIALS AND METHODS
ing interest in finding antioxidants and antimicrobial activity from natural Sample collection - were collected from field and stored in sealed
sources, since they can protect the human body from free radicals and retard polyethyl-ene bags at -20°C until extraction.
the progress of many chronic diseases [1]. Foods rich in antioxidants play an
essential role in the prevention of chronic and degenerative diseases such as Extraction
cardiovascular diseases and cancer [2]. The matured flower petals of Sesbania sesban were soaked in methanol
and acidified methanol (1% conc. hydrochloric acid) separately for 24hrs
Sesbania sesban is a shrub or short-lived tree up to 8 meter tall. Sesbania and the extracted anthocyanin pigments were filtered through Whatmann
sesban can be intercropped with corn, beans, cotton and many other field No.1 filter paper and stored for further analysis.
crops. Harvested leaves make rich compost. Flowers of sesban are known to be
added to stews and omelets in some regions, perhaps mainly as a decora-tive Confirmatory Test For Anthocyanins
element. The extracts had a high content of phenols, flavonoids and S.No Confirmation Test Anthocyanins
anthocyanins . The petals of the flowers are variously shaped and coloured to
attract pollinating insects[3,10].The petal tissues, some or all of these may 1. Heat +2M Hcl for 5min at 100o C Colour stable
possess antibacterial activity as a natural protection system for reproduc-tion 2. Add 2M NaoH dropwise Changes blue to green and slowly fades
3. Visible spectrophotometer Methanol and acidified methanol - 400 to 700nm
and further perpetuation through seed formation.

Therefore, the main objective of the present study was to identify and
determine the phenol, flavonoid, anthocyanin content in Sesbania sesban
flower petals. The other objective was to investigate antimicrobial activity of
extract against common pathogens activities and the antioxidant activity by
hydrogen peroxide scavenging activity and anti-FeCl 2-H2o – stimulated
Determination of total anthocyanin
The total amount of anthocyanin content was determined by using pH
differential method. A spectrophotometer was used for the spectral mea-
surements at 210nm and 750nm (Fuleki & Francis, 1968). The absorbance
of the samples (A) was calculated as follows:
A= [(Absorbance l vis-max-210) pH1.0] – [(Absorbance l vis-max-A750) pH 4.5]

Anthocyanin pigment content (mg/liter) = (A X MW X DF X 1000)/(e X 1)

*Corresponding author. Where,
M.Saravanakumar Molecular weight of anthocyanin (cyd-3-glu) =449g
P.G & Research Department of Extraction coefficient (e) =29,600
Biotechnology,Dr. Mahalingam Center for DF=Diluted factor
research and development, N.G.M.College, Stability at variable pH
pollachi,Tamil Nadu, India The anthocyanin stability was tested by treating 1 ml of sample with 1 ml
of pH 1.0 and 4.5 solutions. The color change was observed .

Journal of Pharmacy Research Vol.5 Issue 1.January 2012 390-293

 M. Kathiresh et al. / Journal of Pharmacy Research 2012,5(1),390-293
Total phenolic assay
Total phenolic compounds in anthocyanin samples were quantified by using
Folin-ciocalteu’s method. About 50 µl of Folin-ciocalteu’s reagent (50% v/v)
were added to 10µl of sample extract. It was incubated for 5min. After
incubation 50µl of 20 % (w/v) sodium carbonate and water was added to final
volume of 400 µl. Blank was prepared by replacing the reagent by water to
correct for interfering compounds. After 30 min of incubation, the absorbance
was measured using spectrophotometer at 760 nm.
Flavonoids confirmation test
A. Ferric chloride: 1 ml of sample extraction was added with a small
amount of Ferric chloride, and results were observed.
B. Aluminium chloride: 1 ml of sample extraction was added with 5% of
Aluminium chloride solution, and results were observed.
Total flavonoid content
The flavonoid content was determined according as the aluminum chloride
colorimetric method described by Chang et al (2002). Aliquots of 0.1ml of
Sesbania sesban extract were taken. To that 0.1 ml of 10 % aluminium
chloride, 0.1 ml of 1 M potassium acetate and 2.8 ml of deionized water were
added. After incubation at room temperature for 40 min, the reaction mixture
was measured at 415 nm absorbance against a deionized water blank on a
spectrophotometer. Quercetin was used as a standard. Using a five point
standard curve (0-50mg/l), the levels of total flavonoid contents in Sesbania
sesban was determined in triplicate, respectively. The data was expressed as
milligram quercetin equivalents (QE)/100 g.
Antimicrobial activity
Sterile Muller Hinton agar medium at around 40ºC was taken and poured in
sterile petriplate and allowed to be solidified, after solidification 9mm wells
were prepared. The organisms used for antimicrobial checking with their
accession number are Escherichia coli (P0096408) ,Klebsiella pnemoniae
(P0098143), Klebsiella oxytoca(P0096408) , Proteus vulgaris (P0096408),
Streptococcus pyogenes(P0098148) ,Pseudomonas aeruginosa(P0096408),
Enterococcus faecalis(P0098146), Staphylococcus aureus(P0096408) , Sta-
phylococcus saprophyticus(P0098153) spreaded over the medium .In these
wells solvent extracts of Sesbania Sesban flower petals were added in different
concentration .The plates was incubated overnight at 37ºC for 24hrs.After
incubation the zones of inhibition were measured. Respective solvent controls
were also run simultaneously.
All analyses were run in triplicate and averaged. BHT and Ascorbic acid
(100mg/ml) solutions were used as standards.
RESULTS
The anthocyanins extracted from Sesbania sesban flower petals using the
solvents methanol and acidified methanol. The UV spectra revealed the
presence of anthocyanin pigments with absorption maxima at 400 to 700nm.
Anthocyanin content were confirmed by ferric chloride test, aluminium
chloride test and also by checking the stability of anthocyanin in extracts with
1M hydrochloric acid and 1M sodium hydroxide (Table 1). Total anthocyanin
content in the methanol and acidified methanol extracts of Sesbania sesban
flower petals were determined using pH differential method. The total
anthocyanin content obtained from the methanol and acidified methanol
extracts were 0.38mg/100g and 0. 28mg/100g respectively.
Table 1.Confirmatory Test For Anthocyanins
S.No Confirmation Anthocyanins Sample In Sample In
Test Methanol Acidified Methanol
1. Heat +2M HCL Colour stable Colour stable Colour stable
for 5min at 100o C
2. Add 2M NaoH dropwise Changes blue to Changes blue Changes blue to
green and slowly fades to green green and slowly fades
3. Visible spectrophotometer Methanol and acidified 433 and 541 430 and 553
methanol -400 to 700nm
The Gallic acid was used as a standard to compare the total phenolic con-
tent of methanol and acidified methanol extracts from Sesbania sesban
flower petal .Using the method of Folin-ciocalteau method the results ob-
tained were 534mg/100g and 180mg/100g respectively. The total
flavonoid content was determined according to the aluminium chloride
colorimetric method. Total flavonoid content in samples were 135mg/100g
and 144mg/ 100g respectively which was compared with the standard
Quercetin. The results showed the composition of Sesbania sesban flower
petal extracts that total phenol and total flavonoid contents were varied
considerably. When compared to anthocyanin and flavonoids, the total
phenol content was found to be higher.
In the preliminary screening, the solvents used are methanol and to obtain
high yield of anthocyanins in the extract, solvents are usually mildly acidi-
fied to facilitate liberation and solubilization of anthocyanins from the
flowers and to stabilize anthocyanins as well. From the stabilized antho-
cyanins in the acidified methanol were used for further experiments.

Analysis for Antioxidant assays Antioxidant Analysis

The antioxidant activity of the Sesbania sesban flower petals of acidified
Hydrogen peroxide scavenging activity methanol extract showed high scavenging activity of 84% at lower
A solution of H2O2 (40mM) was prepared in phosphate buffer (pH concen-tration (1mg) along with the standard BHT (37.65%) and Ascorbic
7.4).The anthocyanin extract (1,10,50,100mg/ml) in 1.6ml phosphate acid is considered as the positive control.
buffer (pH 7.4)was added to a H 2O 2 solution(0.6ml,40mM).The
absorbance value of the reaction mixture was recorded at 230nm.The Hydrogen peroxide scavenging activity
percentage of H2O2 scavenging of anthocyanin extract was calculated Estimation of Anti-FeCl2 –H2O2- Stimulated linoleic acid peroxidation in
using the following for-mula Sesbania sesban flower petals acidified methanol extracts showed a higher
Hydrogen peroxide scavenging activity (% ) = [(Ao-A1)/Ao]*100 inhibition of MDA was 79.9% and comparing with the standard BHT
Ao –the absorbance of the control (77.2%) and Ascorbic acid (87.8%).
A-The absorbance in the presence of the sample of anthocyanin extract Table – 3. Antioxidant activities of anthocyanins extracted from S
esbania sesban flower petals extract
Estimation of Anti-Fecl 2-H202– Stimulated Linoleic Acid Peroxidation
S.No Organisms Accession Concentration Diameter
The effect of anti-Fecl 2-H2O2 – stimulated linoleic acid peroxidation was number of of inhibition
determined by the method as described by Duh(1998) in brief , 0.2ml extract pathogens Zone (mm)
were added to a solution of 0.1 M linoleic acid (0.2ml),2.0mM FeCl 2.4H2 1 Escherichia coli P0096408 - -
O(0.2ml) and 0.2M phosphate buffer (pH 7.4,5.0ml).The reac-tion mixture 2 Klebsiella pnemoniae P0098143 - -
was incubated at 37ºC for 24h.After incubation,0.2ml of BHA 3 Klebsiella oxytoca P0096408 - -
(20mg/ml),1.0ml of thiobarbituric acid (TBA) (1.0%) and 1.0ml trichloro- 4 Proteus vulgaris P0096408 - -
acetic acid (TCA) (10%) were added to the mixture ,which was heated for 5 Streptococcus pyogenes P0098148 - -
6 pseudomonas aeruginosa P0096408 - -
30min in boiling water bath. After cooling, 5.0ml of chloroform was added, 7 Enterococcus faecalis P0098146 - -
and the mixture was centrifuged at 1000rpm to give a supernatant. Absor- 8 Staphylococcus aureus P0096408 12.5mg 5mm
bance of the supernatant was measured spectrophotmetrically at 532nm. 9 Staphylococcus saprophyticus P0098153 12.5mg 2mm

Journal of Pharmacy Research Vol.5 Issue 1.January 2012 390-293

 M. Kathiresh et al. / Journal of Pharmacy Research 2012,5(1),390-293
Antimicrobial activity phenolic and flavonoid contents form Sesbania sesban flower petals.
The antimicrobial activity of the samples are tried with minimum concen- Nuutila et al (2003) reported that the amount of total phenolic varied
tration against Gram-Positive and Gram-negative bacteria using Agar well widely in the Allium extracts.
Diffusion method. The specimen was tested for microbial resistance using
swab technique with an inoculum volume equivalent to 0.5% McFarland’s Antioxidant activity
standard in Muller-Hinton agar and examined after 24hrs. Respective The ability to scavenge hydrogen peroxide could be an efficient
controls(DMSO) were maintained during the investigation. The zone of assessment method to evaluate antioxidant property of plant extracts.
inhibition was found in Gram-Positive bacteria [Staphylococcus Hydrogen per-oxide is a weak oxidizing agent and can inactivate a few
aureus(1mg) and Staphylococcus saprophyticus(12.5mg)] whereas there is enzymes directly, usually by oxidation of essential thiol (-SH) groups. It
no inhibition found in Gram-negative bacteria. can cross cell mem-branes rapidly and inside the cell, H 2O2 probably
Table 2.Antimicrobial activity reacts with Fe2+ and possibly Cu2+ ions to form hydroxyl radical which
may be the origin of many of its toxic effects .It is therefore biologically
S.No Amount of Hydrogen peroxide Anti-Fecl2 -H2 02 – advantageous for cell to control the hydrogen peroxide that is allowed to
Anthocyanins scavenging Stimulated Linoleic accumulate .In this paper, the H2O2- Scavenging activity of the Sesbania
(mg/ml) activity(%) Acid Peroxidation
(%)
sesban extracts along with the standard BHT were determined . The
results indicate that the decrease in this activity was associated with a
1 1 84. 8 79.9 corresponding increase in reducing capacity and scavenging of H 2O2. Duh
2 10 36.2 80.2 et al (1999) in Chrysanthemum morifolium reported results for phenolic
3 50 2 94.1 content and scavenging activity. High antioxidant activity of Red onion
scales[22] and Garlic [25] was reported. Similar results were reported in
The specimen was tested for microbial resistance using swab technique with Carissa carandas and Pergularia daemia root extracts.
an inoculum volume equivalent to 0.5% McFarland’s standard in Muller- It is known that oxidation of poly unsaturated fatty acids in biological
Hinton agar and examined after 24hrs. Respective controls were maintained membranes often lead to the formation and propagation of lipid radicals,
during the investigation . uptake of oxygen, rearrangement of the double membrane lipids. Many of
Control – DMSO these biochemical activities can lead to the production of breakdown prod-
Concentration - 1mg -12.5mg ucts that are highly toxic to most mammalian cell types [28]. Iron salts are
thought to react with H2O2 called the Fenton reaction, to make hydroxyl
radicals, which bring about peroxide reaction of lipids. The effect of ex-
tracts from Sesbania sesban flower petals on the formation of
malonaldehyde (MDA) from linoleic acid .As the concentration of the
antioxidant extracts increased, the formation of MDA decreased. A dose-
dependent MDAinhibition in linoleic acid oxidation was evident. Spinach
showed a higher inhibition of MDA ranging different levels, beyond some
concentra-tion it did not exhibit an inhibitory role wherein at higher
concentrations, the extract is exhibiting a pro-oxidative action.

Fig 1:Staphylococcus aureus
Fig2:Staphylococcus saprophyticus
DISCUSSION
Flowers, fruits and vegetables are good sources of phenolics, flavonoids,
anthocyanins and carotenoids which have health benefits .We presumed that
deep-coloured flowers are phenolic- rich, especially rich in flavonoids.
Generally the flavonoid pigments lies between 400 to 700nm in spectro-
photometer. It was found that considerable amount of phenol , flavonoid and
anthocyanins content in the Sesbania sesban flower petal extracts in methanol
and acidified methanol. The extracts of Cranberry , Roselle(Hibiscus
sabdariffa) and Black currant (Ribena ) commercial juices showed absorption
at the two ranges of wavelength(400 to 700nm). The black-coloured fruit
(Mulberry Fruit, Oriental Plum) red or green colour vegetables (Red Onion,
Beetroot, Ceylon Spinach) seemed rich in total phe-nolic and flavonoids [22]. In
this study , we directly determined the total
Journal of Pharmacy Research Vol.5 Issue 1.January 2012
Antimicrobial activity
As seen from the literature survey that this plant has been mostly studied with
respect to protein inhibitors and antimolluscidal activity from its saponins. Till
date there is no studies have been done on antimicrobial properties of the
Sesbania sesban flower petal extracts. Therefore, this study focuses on the
antimicrobial properties of the flower extracts. A detailed study about
antimicrobial activity of Catharanthus roseus was done by Prajaktra J Patil
(2010). Baydar et al., (2004) reported that acetone:water:acetic acid and ethyl
acetate:methanol:water grape seed ex-tracts inhibited the fifteen bacteria used
as test organisms (Aeromonas hydrophila, B. brevis, B.cereus, B.megaterium,
B.subtilis, E.faecalis, E.coli, Klebsiella pneumoniae, L. monocytogenes,
Mycobacterium smegmatis, Pro-teus vulgaris, P. aeruginosa and S. aureus)
and they attributed the inhibi-tory effect to their phenolic composition. The
grape seed extracts had high total phenolics compared with those of bagasse
(berry without seed and juice), which did not inhibit any of the bacteria tested.
The antimicrobial effects of pomegranate were previously studied. Indeed, it is
reported that the bark, leaves, flowers, and fruits of pomegranate are widely
used as phytotherapeutic agents in Brazil. Ahmad and Beg reported that
alcohol extracts of pomegranate fruits showed antibacterial activity when
tested against S. aureus, E. coli and Shigella dysenteriae. Prashanth et al.
(2001) also reported methanolic extracts of Punica granatum fruit rind to be
active against all microorganisms tested in their study. Mathabe et al., showed
that methanol, ethanol, acetone, and water extracts obtained from pomegranate
were active and effective against the tested microorganisms (S. aureus, E.coli,
Salmonella typhi, Vibrio cholera, S. dysenteriae, S. sonnei, S. flexneri, S.
boydii), showing an inhibition zones of 12-31 mm. Melendez and Capriles
have also reported that extracts from pomegranate fruits pos
390-293
 M. Kathiresh et al. / Journal of Pharmacy Research 2012,5(1),390-293
sess strong in vitro antibacterial activity against many bacteria tested (E.
coli, Enterobacter cloacae, P. fluorescens, Proteus vulgaris , Alcaligenes
faecalis, Serratia marcescens, E.aerogenes, S. aureus, Arthrobacter
globiformis , M. luteus, B. cereus, B. subtilis, B. coagulans,Micrococcus
roseus, M. phlei, M. rodochrus, M. smegmatis ; showing an inhibition
zones of 11-31 mm).
The ethanol extracts of berry and berry skins showed inhibitory effects on
growing of Gram-positive test cultures L. monocytogenes and B. subtilis.Most
wine extracts exhibited some kind of antibacterial activity against gram-
positive and gram-negative strains . In almost all extracts, the diameter of the
inhibition zone for S.aureus was greater than the zone for E.coli,indicating that
the gram-positive strain was more sensitive than the gram-negative one .This
observation can be attributed to differences in the structure of bacteria cell
wall. The less complex structure of the cell wall in the gram-positive bacteria
makes it more permeable to the antimicrobial compounds. The antimicrobial
effects of the tested wine phenolic extracts, especially of the red wines,
indicate that there are phenolic classes which have important bactericidal or
bacteriostatic activities [26].
The antibacterial spectrum of rose petals against the different infectious
pathogenic bacteria indicated that there must be several different
chemicals in the petals of different varieties leading to bacterial inhibition.
The finding of antibacterial activity in rose petals in this study indicates its
ethno botanical evidences on the use of petals of rose flowers to cure
diarrhoeal diseases, opportunistic infection and skin infection [14]. The
phenolic com-pounds are known for their antimicrobial properties. The
significance of these compounds are that these can substitute long term
antibiotic therapy like in case of chronic kidney infection ,bacterial
endocarditis, carrier condi-tions of typhoid(where the organisms resides in
gall bladder).These com-pounds having minimum side effect can be easily
substituted for antibiot-ics. However, if the immune system is too weak or
the organism is too virulent then certain other medication will have to be
given along with these compounds to show significant antimicrobial
activity. In our study the antibacterial property of the Sesbania sesban
flower extract was explained by the zone of inhibition occurred around the
wells containing different concentration of the extracts
REFERENCE
1. Kinsella JE, Frankel E, German, B & Kanner, J (1993) Possible
mechanisms for the protective role of antioxidants in wine and plant
foods. Food Technol. 85–89.
2. Ames BM,Shigena MK , Hagen TM. Oxidants, antioxidants and the
degenerative diseases of aging. Proc. Natl. Acad. Sci. U.S.A. 1993,
90, 7915-7922.
3. Alamaghoul AZ, Bashir, AK, Faronk A and Salih AKM.(1985).
Fitoterapia , 985, 6,331- 337.
4. Chang CC, YangMH,WenHM and Chern JC(2002): Estimation of
total flavonoid content in propolis by two complementary
colorimetic methods .journal of food and drug analysis ,10,178-
182.
5. Cieslik, E., Greda, A & Adamus W. Contents of polyphenols in fruit
and vegetables .Food Chemistry . 2006,94, 135 -142.
6. Fuleki.T and Francis, FJ. Quantitative methods for Anthocyanin;
extraction and determination of total anthocyanin in cranberries J
.Food Sci, 1968,33, 72-77.
7. Harbone, J.B. Comparative Biochemistry of the flavonoids.London
Academic press,1967.
8. Harborne J B and Baxter, H. Phytochemicals dictionary – A hand-
book of bioactive compound from plants .London; Taylor and
Francis Ltd, 1993.
9. J-Y Lin, C-Y Tang. Detemination of total phenolic and flavonoid
contents in selected fruits and vegetables as well as their stimulatory
Source of support: Nil, Conflict of interest: None Declared
Journal of Pharmacy Research Vol.5 Issue 1.January 2012
View publication stats
effects on mouse splenocyte proliferation. Food chemistry, 2007,
101; 140-147.
10. Sesbania Sesban: widely Distributed Multipurpose NFT, NFTA 94-
06, June 1994.
11. Meda A, Lamien CE., Romito M,Millogo J & Nacoulma OG.
Determination of the total phenolic ,flavonoid and praline content in
Burkina Fasan Honey ,as well as their radical scavenging activity
.Food chemistry , 2005 ,91,571-577.
12. Shyamala BN. Leafy vegetables extracts-antioxidants activity and
effecton storage stability of heated oils .Innovative Food science
and Emerging Technologies ,2005,6,239-245.
13. Noureddine Benkeblia . Free –radical scavenging capacity and An-
tioxidant properties of some selected Onions (Allium cepa L.) and
Garlic (Allium sativum L.) Extracts .Brazilian archives of biology
and technology, 2005 ,vol 48, n5,pp.753-759.
14. Hirulkar NB.Antimicrobial activity of rose petals extracts against
some pathogenic bacteria. International journal of pharmaceutical
& biological archives, 2010;1(5):478-484.
15. Williams DH, Stone MJ, Houck PR and Rahman SK ., J.Nat .Prod.,
1989,52,1189-1208.
16. Lcin IG. The antioxidant and radical scavenging activities of black
pepper (piper nigrum) seeds. International .J. Food sciences and
Nutrition, 2005, 56; 491-499.
17. Xiaoyan Zhano,Chao zhang, Claudia guigas,Yue ma ,Margarita
Corrales, Bernhard tausher,Xiaosong hu Composition , antimicro-
bial activity and antiproliferative capacity of anthocyanin ex-tracts
of purple corn (Zea mays L) from China .Eur Food Res Technology ,
2009, 228; 759 -765 .
18. Prajakta J patil, Antimicrobial activity of Catharanthus roseus – A
detail study, British journal of Pharmacology and Toxicology, 2010,
1(1); 40-44.
19. PowersJJ,Somaatmadja D,Pratt DE,Hamdy MK ,1960 ,Food tech-
nology,14:626-632.
20. Duh, P. D. (1998). Antioxidant activity of burdock (Arctium lappa
Linne):Its scavenging effect on free radical and active oxygen.
Journal of the American Oil Chemists’ Society, 75(4), 455–461.
21. Nuutila AM Puuponen-PimiaT., AarniM and Oksman-Caldentey
K.M(2003).Comparsion of antioxidant activities of onion and garlic
extracts by inhibition of lipid peroxidation and radical scav-enging
activity .Food Chemistry ,81,485-493.
22. VeliogluYS, Mazza G, Gao L and Oomah BD (1998).Antioxidant
activity and total phenolics in selected fruits ,vegetables and grain
products J.Agric.Food.chem .,46,4113-4117.
23. Md.Alamgir Hossain, Mohammad S. Rahman, AM. Sarwaruddin
Chowdhury and Mohammad A.Rashid (2007),Bioactivities of
Sesbania sesban Extractives .Dhaka University .J.Pharma.Sci
.,6(1): 61-63.
24. Vijay D. Wagh, Kalpana V.Wagh ,Yogyata N. Tandale, and
Shubhangi A . Salve (2009), Phytochemical,pharmacological and
phytopharmaceutics aspects of Sesbania grandiflora (Hadga): A
review .Journal of Pharmacy Research., 2(5),889-892.
25. Yin , M.: Hwang, S and Chan, K (2002),Nonenzymatic antioxidant
activity of four organosulfur compounds derived form garlic.
J.Agric.Food Chem .,50,6043-6147.
26. Deividas Burdulis, Antanas Äarkinas, Ina Jasutien, Elicija Stacke
vi»Ien, Laurynas Nikolajevas And Valdimaras Janulis , (2009).
comparative study of anthocyanin composition, Antimicrobial and
antioxidant activity in bilberry(vaccinium myrtillus L.) And
blueberry(vaccinium corymbosum L.) Fruits .Acta Poloniae
Pharmaceutica - Drug Research, vol. 66 no. 4 pp. 399-408.
27. Ahmet D. Duman , Mustafa Ozgen , Kenan S. Dayisoylu , Nurcan
Erbil and Coskun Durgac . Antimicrobial Activity of Six Pomegran-
ate (Punica granatum L.) Varieties and Their Relation to Some of
Their Pomological and Phytonutrient Characteristics. Molecules
2009, 14, 1808-1817.
28. Bhaskar and Balakrishnan, (2009).Antioxidant property of
laticiferous plant species.Int.J.Health Res .,2(2):63.
29. Prashanth D, Asha MK, Amit A (2001). Antibacterial activity of
Punica granatum . Fitoterapia, 72: 171-173.
390-293