DPPH Assay

Reagent preparation:

DPPH solution: 2 mg/ml (in methanol)

Extract: 10 mg/ml (in water)

Ascorbic acid: 10 mg/ml (in water)

Instrument used: UV-visible spectrophotometer

Procedure:-

1. All the apparatus were cleaned by using deionized water and dried in hot air oven.

2. All the compounds were weighed and stock solutions were prepared.

3. Dilution of standard, test and blank was prepared. Methanol was used as a solvent.

4. 100 μL DPPH solutions were added to each dilution by using micropipette.

5. Absorbance was taken after 5 minute on UV-visible spectrophotometer at wavelength 517

nm.

Incubation chart:

Sr. No. Reagents Flask A Flask B Flask C

1 Methanol 1950 µL 1900 µL 1900 µL
2 Sample - 50 µL -
3 Ascorbic acid - - 50 µL
4 DPPH 50 µL 50 µL 50 µL

Following formula was used to calculate the % anti-oxidant activity

absorbance of control - absorbance of test

% scavenging activity = x 100
absorbance of control