DPPH test was determined spectrophotometrically this examine depends on the limit of the capacity of the antioxidant to scavenge the radical cation 1,1-dipheny1-2-picrylhydrazyl radical (DPPH). Fifty microlitres of diluted sample (5mg /ml, with solvent) were dissolved in 2 mL of 0.04 mmol/L DPPH in methanol.
A calibration curve in the range 0.05-1.0 mmol/L was used for the Ascorbic acid, and data were expressed as Ascorbic acid equivalent antioxidant capacity (ASAC, mmol/g).
Spectrophoto metric readings were carried out at 517 nm, using plastic cuvette (10 mm) after an incubation period of 60 min in the dark. Samples were analyzed in four replicates and the mean values were calculated