Piper Betel Lab Report

Fresh piper betel leaves were collected from village Sandalpure, Antri of Gwalior, M.P. 1kg of piper betel cut in to small pieces and shade dried. The dried piper betel leaves were ground to a fine powder. 50g of powder was weighed and extracted with soxhelet apparatus using various solvent according to their polarity i.e. petroleum ether, chloroform, methanol, n-butanol, ethyl acetate and water. After solvent extraction, it was evaporated to obtain a powdered extract for various biochemical analysis.

Preliminary phytochemical screening of the extracts was performed for the presence of alkaloids, flavonoids, steroids, tannins, saponin, phenol and by the standard procedures.

Alkaloids: To 1 ml of extract, 2-3 drops of Wagner’s reagent were

To each sample solution (1.0 ml) and the standard (gallic acid) was added 5 ml of Folin-Ciocalteu (sigma-aldrich) and 4 ml sodium carbonate (7% w/v) and shaken. The solution could stand for 30 min in the dark at room temperature, after which absorbance was measured at 765 nm using a spectrophotometer. The phenolic content was calculated from the standard curve of gallic acid15.

Total Flavonoid Content

A known volume of extract was placed in a 10ml volumetric flask add distilled water to make final volume 5 ml followed by adding 0.3 ml NaNO2(1:20). Add 3ml AlCl3 (10%) 5 min later. After 6 min, 2 ml 1 M NaOH was added and the total volume was made up to 10ml with distilled water. The solution was mixed well again, and the absorbance was measured against a blank at 510 nm with a (UV-VISIBLE Parkin Elmer Lambda 23 with win lab N6.0software.). The flavonoid content was calculated with quercetin as standard16.

Reducing Power

To 200 µl sodium nitroprusside (5Mm), 800 µl extracts (0.1-1 mg/ml) dissolved in PBS (25 mM, pH 7.4)was added.The mixture was incubated for 2.5 hrs. at 37ºC under normal light followed by incubation in dark for 20 min. 600 µl Griess reagent was added and incubated for 40 min. at room temperature and absorbance was measured at 540 nm against a suitable blank (2ml H2O and 0.6 ml Griess reagent). Control (1.6 ml H2O, 400µl SNP and 600µl Griess reagent) was prepared and percent of inhibition was calculated by using this