DPPH Antioxidant Activity Evaluation

The antioxidant activity of n-hexane, chloroform extract, and isolated compounds was measured according to published using methods using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) with help of UV spectrophotometry [24 (link)]. 18 mg of each extract was dissolved in 100 ml methanol to prepare a stock solution of the extract similarly 2 mg of isolated compounds were dissolved in 100 ml methanol to prepare the stock solution of the compound. 9.5 mg of DPPH was dissolved in 25 ml methanol to prepare a 1 Mm Solution of DPPH. From the Stock solution, different concentration of the solution is prepared such as 5, 10, 30, 50, and 100 μg/ml of extract and isolated compound with help of the dilution formula. Now 1 ml of DPPH solution was added to each sample as well as the control and kept for 30 min in dark. After 30 min the absorbance was measured for each sample at 517 nm by using UV spectrophotometry. The decrease in the DPPH absorbance indicates the increase in 2,2-Diphenyl-1-picrylhydrazyl scavenging activity. Free radical scavenging by DPPH defines the % radical scavenging. The % antioxidant effect was calculated with help of the below formula. A0 = Control absorbance; A1 = Absorption of sample.