QUALITATIVE PHYTOCHEMICAL ANALYSIS AND …

1 , (1)2015:81-84 ISSN2278 910381 QUALITATIVE PHYTOCHEMICAL ANALYSIS AND estimation OFTOTAL PHENOLS AND FLAVONOIDS IN LEAF EXTRACTOFSARCOCHLAMYS PULCHERRIMAR ajesh Kumar Shah1*& Yadav21 Dept. of Zoology, College,Dibrugarh, Assam2 Dept. of Life Sciences, Dibrugarh University,Dibrugarh, Assam*Corresponding author s medicinal properties of plants are due to some chemical substances that produce certain definite physiological actionon the human body. These non-nutritive components are called QUALITATIVE ANALYSIS as well asquantification of phytochemicals of a medicinal plant is regarded as vital step in any kind of medicinal to family Urticaceae, leaves which has long been used by some tribal people ofAssam as food and to treat several shoots, leaves and fruits are eaten as vegetable especially bytheMishing tribe with pork; based on thestrong believe that it kills the tap are used for diarrhea anddysentery, as carminative and digestiveand there is no scientific evidence for above said activities.

2 The present study wascarried out to test the presence of various phytochemicals in the leaf extract of the plant and also to estimate the totalphenol and flavanoids in the leaf ANALYSIS of the leaf extract ofSarcochlamyspulcherrimarevealedthe presence most of the biochemicals tested for such ascarbohydrate,protein,alkaloid, tannin, flavanoid, Steroids,terpenoids,phenol, saponin and total phenolic content of the methanolic leaf extract is found to + mg/gand the total flavonoid contents of the leaf extract is found to be + presence of variousphytochemicals in the tested plant reveals that this plant may be a goodsource for production of new drugs for WORDS:Sarcochlamyspulcherrima,Phytochemi cal ANALYSIS ,Total Phenols and Flavonoids, DibrugarhINTRODUCTIONM edicinal plants are being used by human sinceimmemorial time.

3 The medicinal properties of plants aredue to some chemical substances that produce certaindefinite physiological action on the human body. Theseare non-nutritive chemicals that have protective orpreventive power against phytochemicals aregrouped into two main categories1, primary constituentswhich includes amino acids, common sugars, proteins andchlorophyll etc., and secondary constituents consisting ofalkaloids, essentialoils, flavonoids, tannins, terpenoids,saponins, phenolic compounds etc. (Krishnaiahetal.,2009and Edeogaet al.,2005) .The QUALITATIVE ANALYSIS as wellas quantification of PHYTOCHEMICAL constituents of amedicinal plant is considered to be a vital step in any kindof medicinal plant to family Urticaceae,leaves which has long been used by some tribal people ofAssam as food and to treat several shoots,leaves and fruits are eaten as vegetablesespecially bytheMishing tribe with porkbased on thestrong believe thatit kills the tap are used fordiarrhea anddysentery, they are carminative and digestiveand there isno scientific evidence for above said activities.

4 Thepresent study was carried out to test the presence ofvarious phytochemicals in the leaf extract of the plant andalso to estimatethe total phenol and flavanoids in the & of Plant MaterialFresh leaves ofSarcochlamyspulcherrimawere collectedfrom different localities of Dibrugarh, Assam and washedproperly for removal of dust leaves wereshade dried and crushed using blender. The dry powderthus obtained was stored in air tight of ExtractCrude leaf extract was prepared by using Soxhletapparatus. About 30g of finely powdered leaves werefilled in a thimble and were extracted using differentsolvents (250 ml) such as Methanol, Ethanol andPetroleum ether for about 48 hours.

5 The extracts wereconcentrated by heating on a hot plate at about 30 -40 Cand preserved at 40C until use. The aqueous extract wasprepared by boiling 10g of dried leaf powder in 300 ml ofwater for about 30 minutes over a hot pan at 30-400C. Theextract was filtered, concentrated and then stored at 40 Cfor TestsPhytochemical ANALYSIS was done following standardmethods(Sadasivam and Manickam 1996,Tyler 1994 andHarborne 1973).a)Test for CarbohydrateMolisch s Test:To 2ml of extract, 3-4 drops of Molisch sreagent was added and mixed properly. To thisconcentrated Sulphuric acid was added by the walls of theAnalysis and estimation of leaf extract ofSarcochlamyspulcherrima82test tube.

6 Appearance of a purple or blue ring in betweenthe two layers indicates the presence of )Test for ProteinBiuret Test:2ml of biuret reagent (mixture of 2ml of 10%NaOH and 2-3 drops of CuSO4)was added to thecrude extract and heated. Appearance of purple/bluecolour confirms the presence of Test:2 ml of extract was treated with %Ninhydrinand heated for 5-10 minutes. Blue colourindicates the presence of )Test for AlkaloidMayer s Test:The crude extracts were evaporated todryness and residues were heated with 2% Hydrochloricacid on a boiling water bath. The extract were cooled,filtered and treated with the Mayer s reagent.

7 Presence ofyellow precipitation or turbidity shows the presence )Test for PhenolFerric chloride test:To 2ml of plant extract, 2ml ofdistilled water followed by 10 % FeCl3solution wasadded. Bluish black colour indicates the presence )Test for SaponinFoam Test:2 ml of extract was taken in a test tube and 10ml of distilled water was added and shaken of foams confirms the presence of )Test for TanninsGelatin Test:Crude plant extracts were treated with 5 mlof 1% gelatin solution containing NaCl and observed forthe occurrence of white chloride Test:2 ml of 5 % FeCl3solution wasadded to 2 ml of plant extract.

8 Appearance of dark blue ofgreenish black colour indicated the presence of )Test forFlavanoid4 ml of extract was taken and about 2ml of50% methanolwas added. The solution was warmed and metalmagnesium was added. This was followed by addition of 5to 6 drops of concentrated hydrochloric acid. Redcoloration confirms the presence of )Test for GlycosidesLibermann s Test:To the crude extract, 2ml of chloroformand 2 ml of acetic acid was added. The solution was icecooled followed by addition of conc. H2SO4. Colourchange from blue to green indicates the presence s Test:Crude extract was dissolved in 2 ml ofchloroform.

9 To this conc. H2SO4was added and themixture was shaken. Formation of reddish brown colourindicates the presence of Test:To the crude extract was added 2mlofacetic acid and few drops of 2 % FeCl3solution. The entiremixture was then poured in a test tube containing 2 ml ofconc. brown ring at the junction indicates thepresence of )Test for SteroidsSulphuric acid Test:To the plant extracts 2 ml ofchloroform was added. 2 ml of conc. H2SO4was added bythe sides of the test tubeand observed for red colour at thelower chloroform )Test for TerpenoidsSulphuric acid Test:Crude plant extract was dissolved in3ml ofchloroform.

10 This was than evaporated to drynessand 2ml of conc. H2SO4was added and heated for about3minutes. A grayish colour indicated the presence of Total PhenolPhenol content in the plant extract was determined byFolin-Ciocalteu reagent method with slightmodificationsAdedapoet al.,2009; Koncicet al.,2001;McDonald et al., 2001 and Nabaviet ). One gramof the sample was extracted with 10 ml of 80% homogenate was centrifuged at 10,000 rpm for 20minutes. The supernatant was collected and evaporated solution of the extract in theconcentration of 1 mg/ml was used in the ANALYSIS . Thereaction mixture was prepared by mixing ml ofmethanolic solution of extract, ml of 10%Folin-Ciocalteu s reagent dissolved in water and ml Blank was concomitantly prepared, ml methanol, ml 10% Folin-Ciocalteu s reagentdissolved in water and ml of of standard curve was prepared usinggallic acid.