Simultaneous detection of the antioxidant and pro-oxidant activity of dietary polyphenolics in a peroxidase system

The ability to reduce the peroxidase (myeloglobin/H2O2)-generated ABTS(.+) [2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) radical cation] has been used to rank the antioxidant activity of various agents including dietary flavonoids and chalcones. Surprisingly, we found that in the presence of catalytic concentrations of the phenol B-ring containing flavonoids, apigenin, naringenin and the chalcone phloretin, the formation of the ABTS(.+) was initially increased. The enhanced formation of the ABTS(.+) was attributed to the peroxidase/H2O2 mediated generation of polyphenolic phenoxyl radicals that were able to co-oxidize ABTS. The relative ABTS(.+) generating ability of these dietary polyphenolics correlated with their ability to co-oxidize NADH to the NAD* radical with the resultant generation of superoxide. This pro-oxidant activity was not observed for either luteolin or eriodyctiol, which are B-ring catecholic analogues of apigenin and naringenin, respectively, suggesting that these antioxidants are incapable of the transition metal-independent generation of reactive oxygen species. This pro-oxidant activity of the polyphenolics therefore needs to be taken into account when quantifying antioxidant activity.