The principle of the ABTS method for determining the T-AOC is as follows. ABTS is oxidized to green ABTS+ by appropriate oxidant, which can be inhibited if there exist antioxidants. The T-AOC of the sample can be determined and calculated by measuring the absorbance of ABTS+ at 414 nm or 734 nm. Trolox is an analog of VE and has a similar antioxidant capacity to that of VE. Trolox is used as a reference for other antioxidant antioxidants. For example, the T-AOC of Trolox is 1, then the antioxidant capacity of the other substance with the same concentration is showed by the ratio of its antioxidant capacity to Trolox antioxidant capacity.

If the sample to be tested is water-soluble, dilute the standard and samples with PBS. If the sample to be tested is water-insoluble, dilute the standard and samples with 80% ethanol.

                                   Note: A：blank wells, B-H：standard wells, S1-S80：sample wells.

1)     Standard tube: Add 10 μL of standard with different concentration to the 96 well micro-plate.

        Sample tube: Add 10 μL of sample to the 96 well micro-plate.

2)     Add 20 μL of reagent 4 application solution to each well.

3)     Add 170 μL of ABTS working solution to each well.

4)     Mix fully and stand for 6 min at room temperature. Measure the OD values of each well at 414 nm with Microplate reader.