5 ml extract solution and observed for white precipitation which indicates presence of tannin. 0.2 g of the extract was Modulators shaken with 5 ml of distilled water and then heated to boil. Frothing shows the presence of saponin. 0.2 g of the extract was dissolved in 10% NaOH solution, yellow colouration indicates the presence of flavonoid. To 2 ml of extract solution, added 2 ml of alcohol and few drops of ferric chloride solution and observed for colouration. Five ml of each extract was treated with 2 ml of glacial acetic acid containing one drop of ferric chloride solution. This was under layered with 1 ml of conc. sulphuric selleck screening library acid. A brown ring at the interface indicated

the present of cardiac glycoside. (A violet ring may appear below the ring while in the acetic acid layer, a greenish ring may formed). 0.5 g extract was boiled with conc. HCl and filtered. 0.5 ml of picric

acid and Mayer’s reagent was added separately to about 1 ml of the filtrate in a different test tube and observed for coloured precipitate or turbidity. To 0.2 g of extract, added 5 ml of chloroform and 5 ml of 105 ammonia solution. The presence of bright pink colour in the aqueous layer indicated the presence of anthraquinone. Five ml of extract solution was mixed in 2 ml of chloroform, and 3 ml of conc. sulphuric acid was added to form a layer. A reddish brown colouration of the interface buy BVD-523 was formed to indicate the presence of terpenoids. Red colour at the lower surface indicates presence of steroid. To 0.5 ml of extract solution, 1 ml of water and heated after adding 5–8 drops of Fehling’s solution. Brick red precipitation indicated the presence of reducing sugar. Antioxidants react with 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical and convert it to 1, 1-diphenyl-2-picryl hydrazine. The degree of change in colour from purple to yellow can be used as a measure of the scavenging potential of antioxidant extracts. Aliquots of ethanol extract solutions (1 mg/ml) were taken and made up the volume to 3 ml with methanol. 0.15 ml of freshly prepared DPPH 3-mercaptopyruvate sulfurtransferase solution

was added, stirred and left to stand at room temperature for 30 min in dark. The control contains only DPPH solution in methanol instead of sample while methanol served as the blank (negative control). Absorbance was noted at 517 nm using the Systronics make spectrophotometer (Visiscan 167). The capacity of scavenging free radicals was calculated as scavenging activity (%) = [(Abscontrol−Abssample/Abscontrol)] × 100 where Abscontrol is the absorbance of DPPH radical + methanol; Abssample is the absorbance of DPPH radical + sample extract/standard. The ABTS assay was carried out following the method of Re et al.9 The stock solution included 7 mM ABTS solution and 2.4 mM potassium persulfate solution and mixed them in equal proportion then allowed to react for 12 h at room temperature in the dark and diluted by mixing 1 ml ABTS solution with 60 ml methanol to obtain an absorbance of 0.706 ± 0.