Evaluation Of Antioxidant Activities Of Flower Extract .

Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 251-259ISSN: 2319-7706 Volume 3 Number 4 (2014) pp. 251-259http://www.ijcmas.comOriginal Research ArticleEvaluation of antioxidant activities of flower extract (fresh and dried)of Saraca indica grown in West BengalTapan Kumar Pal1*, Sauryya Bhattacharyya2 and Ankita Dey21Department of Biotechnology, Bengal Institute of Technology, On Basanti Highway, Hadia,Kolkata-700150, West Bengal, India,2Department of Food and Nutrition, Sarada Ma Girls College, Nabapally, Barasat, Kolkata700126, West Bengal, India*Corresponding authorABSTRACTKeywordsSaraca Ascorbic acid;DPPH radicalscavengingactivity.Saraca indica has been greatly used as traditional medicine for women relatedproblems, such as leucorrhoea, menorrhagia, dysfunctional uterine bleeding,bleeding haemorrhoids etc. In this study different Phytochemicals and free radicalscavenging activity were measured in the ethanolic and water extract of fresh anddried flowers of Saraca indica collected from in and around Barrack pore area,West Bengal. The Phytochemicals studied from flower extracts are totalpolyphenols, flavonoids, ascorbic acid and tannins. Free Radical scavengingactivities of the extracts were evaluated using DPPH assay method. The resultsrevealed that total polyphenols, flavonoids and tannins content were relativelyhigher in ethanolic extract of the fresh flower and water extract of the dried flowerof Saraca indica. Whereas both extract of dried flower contained higher amount ofascorbic acid. The free radical scavenging activity was higher in fresh flowerextract (both ethanol and water) in comparison to dried flower. The results revealedthat the antioxidant property of both flower extracts may improved human healthstatus and stay away from many diseases.Introductionreactive species contribute to most chronicdiseases. It is hypothesized thatantioxidants originating from foods maywork as antioxidants in their own right invivo, as well as bring about beneficialhealth effects through other mechanisms,including acting as inducers ofmechanisms related to antioxidant defenseMost bioactive food constituents arederived from plants; those so derived arecollectively called phytochemicals. Thelarge majority of these phytochemicals isredox active molecules and thereforedefined as antioxidants. Antioxidants caneliminate free radicals and other reactiveoxygen and nitrogen species, and these251

Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 251-259(Kensler et al., 2007; Jeong et al., 2006),longevity (Baur et al., 2006; Wood et al.,2004), cell maintenance and DNA repair(Astley et al., 2004).West Bengal, India during the month ofJuly to August, 2013.Saraca indica is highly regarded as auniversal panacea in the ayurvedicmedicine. It is one of the universal planthaving medicinal activities. Saraca indicahas been greatly used as traditionalmedicine for women related problems,such as leucorrhoea, menorrhagia,dysfunctional uterine bleeding, bleedinghaemorrhoidsetc(ayurvedicpharmacopoeia of India. 2001).Theantimicrobial activity of the stem and barkof Saraca indica have been evaluatedagainst standard strain of Staphylococcusaureus, Escherichia coli, Salmonellatyphimurium (Shilpakala Sainath et al,2009). The leaves of Saraca indica alsoevaluated for anthelmintic activity(Manjunath et al, 2006; Nayak et al,2011), analgesic and antipyretic activities(Pradhan et al, 2010), CNS depressantactivity (Yadav et al, 2008).Folin-Ciocalteu reagents, 1, 1-diphenyl-2picrylhydrazyl (DPPH) reagent, gallicacid, aluminium trichloride (AlCl3),butylated hydroxytoluene (BHT), catechinand 2, 6-dichloroindophenol (DCIP) werepurchased from SRL India, Sulphric acid,Sodium nitrates (NaNO2), sodiumhydroxide (NaOH), sodium carbonate(Na2CO3) were purchased from Merck(India). Double distilled water was usedfor the complete study.The reports of quantitative estimation ofdifferent antioxidants of the flower ofSaraca indica are hardly available. Theantioxidant property is also related to thecondition of soil and environment wherethe plant is grown. So in this investigationwe are quantitatively estimate differentphytochemicals such as total polyphenols,flavonoids, ascorbic acid and tannins andfree radical scavenging activities ofDPPH to evaluate antioxidants propertiesof the flower of Saraca indica.Ethanol and water extracts from Driedand Fresh FlowersChemicals:Preparation of extracts:The flowers were collected and healthyflowers were shade dried and thenpowdered using electric blender to get acoarse powder. The Fresh whole offlowers was also used in these studies. Allthe plant materials were stored at 4oC forfurther use.The ethanol and water extracts of driedand fresh flowers were prepared bymortar-pestle using respective solvents(water and ethanol) separately andgrounded paste of flowers with solventwere then shake for 24 h in a shaker. Theextracts were filtered through glass wool.The extraction process was repeated twice.The collected filtrates were dried at roomtemperature and weighted by electronicsbalance and diluted by distilled water to adesiredconcentration(10 g/ml)(Maneemegalai and Naveen, 2010).Theses extracts were stored in refrigerators(4oC) for further used.Materials and MethodsPlant materialThe fresh flowers of Saraca indica werecollected from in and around Barrackpore,252

Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 251-259acid was used for constructing thestandard curve (20 to 100 g/ml;) and thetotal phenolic compounds concentration inthe flower extract was expressed asmilligrams of gallic acid equivalent pergram of dry weight (mg GAE/g) ofextract.Antioxidantmeasurementassaymethods: All the experiments wereperformed in triplicate using the followingprocedure:1, 1-diphenyl-2-picrylhydrazyl (DPPH)radical scavenging assayThe antioxidant activities of extracts offresh flower and dried flower powder wereassessed by method reported bySasidharan et al. (Sasidharan et al, 2007)with some modification. 0.002% DPPHwas prepared in ethanol. 250 µl of DPPHsolution was mixed with 5 µl of flowerextracts and final volume of 1000µl wasmade up by adding ethanol. The mixtureswere kept in dark for 20 min and opticaldensity was measured at 517 nm usingSpectrophotometer (Systronics make,Model no 2202) ethanol (750µl) withDPPH solution (250 µl) was used ascontrol. The percentage of inhibition ofDPPH activity was calculated (Chorage etal, 2013) using the formula given below:Percent of inhibition of DPPH activity (absorbance of controlTotal Flavonoid content assayTotal flavonoid content of the flowerextract was determined according tocolorimetric method described by Zhishenet al. (Zhishen et al., 1999), with somemodification. Briefly 0.5 ml sample wasmixed with 2 ml of distilled water and0.15 ml of sodium nitrite (NaNO2, 5%w/v), allowed to stand for 6 min, 0.15 mlaluminium trichloride (AlCl3, 10% w/v)was added and allowed to stand again for6 min, followed by addition of 2 ml ofsodium hydroxide (NaOH, 4% w/v). Thefinal volume was make up to 5 ml bydistilled water. The reaction mixture wasmixed thoroughly and allowed to stand foranother 15 min. The absorbance of pinkcolour that developed was measured at510nmusingspectrophotometer(Systronics make, Model: 2202). Distilledwater was used as blank. All theexperiment was carried out in triplicate.The total flavonoid content was expressedin mg of catechine per gram of flowerextract.absorbance of sample) 100Absorbance of controlTotal phenolic content assayThe total phenolic content was measuredusing Folin-Ciocalteus reagent based onprocedures described by Singleton et al.(Singleton et al., 1999), with somemodifications. Briefly, 0.5 ml of samplewas mixed with 1.5 ml (1:10 v/v dilutedwith distilled water) Folin-Ciocalteau sreagent and allowed to stand for 22 C for5 min. Then 2 ml of sodium carbonate(Na2CO3, 7%, w/v) was added and themixture were allowed stand for another 90min and kept in the dark with intermittentshaking. Then the absorbance of the bluecolour that developed was measured at725nmusingspectrophotometer(Systronics make, Model no 2202). GallicTannin assayContent of Tannins in the flower extractwas determined by Folin Denis method(Polshettiwar and Ganjiwale, 2007).Briefly 1 ml of sample or standard solutionof Tannic acid (5µg/ml - 40µg/ml) wasmixed with 0.25 ml Folin Denis reagentand 0.50ml saturated Na2CO3 solutionware added to it. The volume was made upto 5 ml with distilled water and absorbance253

Int.J.Curr.Microbiol.App.Sci (2014) 3(4): 251-259was measured at 700 nm after 30 min ofincubation. The total tannic acid contentwas expressed as mg of tannic acidequivalent per gram of dry weight of thesample (Kalpana et al, 2013)Results and DiscussionPhytochemicals and antioxidant activityanalysis of water and ethanol extract ofSaraca indica fresh flowers and driedflowers were given in Table.1 and Table.2respectively.Vitamin C assayVitamin C of the flower extract wasmeasured titrematrically by 2, 6dichloroindophenol (DCIP) solution. The2, 6-dichloroindophenol (DCIP) solutionwas prepared by dissolving 52 mg of 2, 6dichloroindophenol in about 500 mL ofwater. Sodium bicarbonate (42 mg) is thenadded and dissolved. The resultingsolution is finally diluted to 1 L withdistilled water.Total polyphenol contents (mg/g offlowers) of ethanolic extract of fresh anddried flowers of Saraca indica were 4.509mg/g and 3.146 mg/g respectively(Figure.1). Where as in the water extractof fresh and dried flowers, totalpolyphenol contents were 1.068 mg/g and2.190 mg/g respectively. From theseobservations, we can say that totalpolyphenol content in ethanolic extract isrelatively 46.31% higher in fresh flowersthen dried flowers. But the water extract ofdried flowers contain 105.05% more oftotal polyphenol than fresh flowers.Briefly 5.00 mL of the sample or standardascorbic acid solution was taken into a250-mL Erlenmeyer flask, 2 mL of the 3%Metaphosphoric acid mixture and about 25mL of distilled water to the flask wasadded. This mixture was then titrated withthe DCIP solution until a permanent(lasting more than 30 sec) light red or pinkcolour appears. The volume of DCIPneeded to oxidize the sample and standardascorbic acid was correlated to find outascorbic acid content in the sample. Theresult was expressed in mg of ascorbicacid per gram of extract.Total ascorbic acid contents (mg/g offlowers) of Saraca indica flowers are alsopresented in the Figure 2. Ethanol extractof fresh flowers contained ascorbic acid0.124mg/g and dried flowers contained0.426 mg/g. Whereas water extract offresh flowers and dried flowers containedascorbic acid 0.113 mg/g and 0.415 mg/grespectively. Therefore from the aboveresults it can be seen that ascorbic acidcontent of ethanolic and water extract ofdried flowers are 235.48% and 267.25%respectively higher than fresh flowers.Statistical analysisAll the analysis were carried out intriplicate and expressed as mean SD.Analyses of variance were performedusing the one-way analysis of variance(ANOVA).Significantdifferencesbetween means were determined byDuncan s multiple range tests. P valuesless than 0.05 were considered statisticallysignificantTotal tannin content (mg/g of flowers) ofSaraca indica flowers is presented in theFigure 3. Ethanol extract of fresh flowerscontained tannin 0.720mg/g and driedflowers contain 0.486mg/g, whereas waterextractof fresh and dried flowers254