An interlaboratory comparison of methods used to assess antioxidant potentials1

Abstract

Many analytical methods are used to measure the antioxidative activity of substances yet little is known about the comparability of the test results between laboratories. After an initial evaluation of a broad range of methods conducted by one laboratory, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, the trolox equivalent antioxidant capacity (TEAC) assay, the lipid assay (or 2,2'-azobis(2-aminepropane) (ABAP) assay) and the thiobarbituric acid (TBA) assay were selected to be evaluated in the interlaboratory study. The antioxidative potentials of trolox, tocopherol, lipochroman-6, ascorbic acid, 4-methyl-brenzcatechin, and/or 3,5-di-tert-butyl-4-hydroxytoluene (BHT) were assessed using each of the methods. These methods were then evaluated in respect of their reproducibility and classification properties. Based on the results of this study, the DPPH assay followed by the TEAC assay yielded the best results based on reproducibility and sensitivity both within one laboratory and between laboratories. The results of the interlaboratory study were then compared with the single center results obtained from the commercially available photochemolumiescence (PCL) kit. To assess the transferability of chemical data to biological systems, they were also compared with the single center results obtained using the cell-based Dichlorodihydrofluoresceine (DCFH) assay.