DPPH (2,2-Diphenyl-1-picrylhydrazyl) is a stable free radical that can be used to measure the radical scavenging activity of antioxidants. The odd electron of nitrogen atom in DPPH is reduced by receiving a hydrogen atom from antioxidants to the corresponding hydrazine. DPPH method may be utilized in aqueous and nonpolar organic solvents and can be used to examine both hydrophilic and lipophilic antioxidants^[1]. In Vitro: DPPH shows a strong absorption band at 517 nm due to its odd electron and solution appears a deep violet colour, the absorption vanishes as the electron pairs off. The resulting decolorization is stoichiometric with respect to the number of electrons taken up. The alcoholic solutions of 0.5 mM are densely colored and at this concentration, the Lambert-Beer law is obeyed over the useful range of absorption^[1].
DPPH assay is a rapid, simple, inexpensive and widely used method to measure the ability of compounds to act as free radical scavengers or hydrogen donors, and to evaluate antioxidant activity of foods. It can also be used to quantify antioxidants in complex biological systems, for solid or liquid samples. This method is easy and applies to measure the overall antioxidant capacity and the free radical scavenging activity of fruit and vegetable juice. This assay has been successfully utilized for investigating antioxidant properties of wheat grain and bran, vegetables, conjugated linoleic acids, herbs, edible seed oils, and flours in several different solvent systems including ethanol, aqueous acetone, methanol, aqueous alcohol and benzene. It is a convenient method for the antioxidant assay of cysteine, glutathione, ascorbic acid, tocopherol and polyhydroxy aromatic compounds, for olive oil, fruits, juices and wines^[1].