Applicability of an improved Trolox equivalent antioxidant capacity (TEAC) assay for evaluation of antioxidant capacity measurements of mixtures.

The TEAC (Trolox equivalent antioxidant capacity) assay is based on scavenging of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) radical anions (ABTS(.-)). In this report we describe a modification based on pre-generation of the ABTS radical anions with a thermolabile azo compound, 2,2'-azobis- (2-amidinopropane)HCl (ABAP). This modification makes the assay less susceptible to artefacts, e.g. influence on the radical generation process. For most antioxidants tested, a biphasic reaction pattern was seen, i.e. a fast and slow scavenging rate. We evaluated application of the assay with both lipophilic and hydrophilic compounds with antioxidant capacity. Several organic solvents, compatible with water, were tested with alpha-tocopherol, quercetin and beta-carotene. It was found that the TEACs differed in various solvents. Under standardized conditions additivity of TEACs obtained from individual antioxidants could be demonstrated. This might enable application of the assay for the identification of "unknown" antioxidants.