Antioxidant Activity using (FRAP) Method

Determination of total antioxidant activity (FRAP assay) for plant extract by modified method of Benzie and Strain.

1. The stock solutions included 300mM acetate buffer (3.1g C2H3NaO2·3H2O and 16 ml C2H4O2), pH 3.6, 10mM TPTZ (2, 4, 6-tripyridyl-s-triazine) solution in 40mM HCl, and 20mM FeCl3 6H2O solution.
2. The fresh working solution was prepared by mixing 25ml acetate buffer, 2.5ml TPTZ, and 2.5ml FeCl3 6H2O.
3. The temperature of the solution was raised to 37°C before use.
4. Samples (150μL) were allowed to react with 2850μl of the FRAP solution for 30 min in the dark condition. Readings of the colored product (ferrous tripyridyltriazine complex) were taken at 593 nm.
5. The standard curve was linear between 200 and 1000μM FeSO4.

Results are expressed in μM