Formation and decay of the ABTS derived radical cation: A comparison of different preparation procedures

Abstract Bleaching of a preformed solution of the blue‐green radical cation 2,2′‐azinobis (3‐ethylbenzothizoline‐6‐sulfonic acid) (ABTS +· ) has been extensively used to evaluate the antioxidant capacity of complex mixtures and individual compounds. The reaction of the preformed radical with free‐radical scavengers can be easily monitored by following the decay of the sample absorbance at 734 nm. The ABTS radical cation can be prepared employing different oxidants. Results obtained using MnO 2 as oxidant show that the presence of manganese ions increases the rate of [ABTS] +· autobleaching in a concentration‐dependent manner. The radicals can also be obtained by oxidizing ABTS with 2,2 ′ ‐azobis(2‐amidinopropane)hydrochloride (AAPH) or peroxodisulfate (PDS). The oxidation by AAPH takes place with a large activation energy and a low reaction order in ABTS. The data support a mechanism in which the homolysis of AAPH is the rate‐limiting step, followed by the reaction of ABTS with the peroxyl radicals produced after the azocompound thermolysis. On the other hand, the low activation energy measured employing PDS, as well as the kinetic law, are compatible with the occurrence of a bimolecular reaction between the oxidant and ABTS. Regarding the use of ABTS‐based methodologies for the evaluation of free radical scavengers, radical cations obtained employing AAPH as oxidant can be used only at low temperatures, conditions where further decomposition of the remaining AAPH is minimized. The best results are obtained with ABTS derived radicals generated in the reaction of PDS with an ABTS/PDS concentration ratio equal (or higher) to two. However, even with radicals prepared by this procedure, stoichiometric coefficients considerably larger than two are obtained for the consumption of the radical cation employing tryptophane or p ‐terbutylphenol as reductants. This casts doubts on the use of ABTS‐based procedures for the estimation of antioxidant capacities. © 2002 Wiley Periodicals, Inc. Int J Chem Kinet 34: 659–665, 2002