The 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation can be generated by incubation of ABTS and 2,2'-azo-bis(2- amidinopropane) at 45 degrees C. The ABTS radical cation is stable for several minutes at room temperature and reacts quantitatively and instantaneously with several antioxidants, such as Trolox, ascorbic acid, uric acid, cysteine, glutathione and bilirubin. In contrast, the ABTS radical cation reacts slowly with albumin. When serum is added to a solution of the ABTS radical cation, the bleaching of the radical follows biphasic kinetics, with a fast decay followed by a slow decay that takes place within several minutes. The fast decay is primarily due to uric acid, while the slow decay is related to the protein content of the sample. We propose that this procedure can provide an independent and simultaneous evaluation of the low molecular weight and protein antioxidants present in biological samples such as serum.