![]() ![]() A common use for it is in the enzyme-linked immunosorbent assay (ELISA) to detect the binding of molecules to each other. The present study makes a recommendation for estimating the ABTS*+ capacity accurately and reasonably. In biochemistry, ABTS (2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) is a chemical compound used to observe the reaction kinetics of specific enzymes. However, full and accurate evaluation of antioxidant reactivity rather than capacity requires recording ABTS*+ loss continuously during the whole reaction period. ABTS*+ assay could be measured within 2-10 min to obtain a rough result, which was mostly 6 min in the literature. When using different pre-diluting solvents, different detection wavelengths should be selected. Co-based metal organic frameworks (ZIF-67), working as an oxidase-mimicking nanozyme, can simultaneously catalyze the oxidation of 3,3,5,5-tetramethylbenzidine, 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Amplex red, exhibiting a high catalytic activity. The results showed that the solvent effects were very significant. To develop an effective and universal method for estimating the ABTS*+ capacity accurately and reasonably, five pre-dilution solvents and different reaction times were investigated in ABTS*+ assays of five typical antioxidants. The existing methods for ABTS*+ radical-scavenging activity assays are diverse in pre-diluting solvents and reaction time, which lead to errors in the TAC estimations. 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS*+) is a stable free radical frequently used for estimating the total antioxidant capacity (TAC) of natural products. ![]()
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