Evaluation of chemiluminescence reagents for selective detection of reactive oxygen species

In order to evaluate the chemiluminescence (CL) reagents for selective detection of reactive oxygen species (ROS), we comprehensively measured the CL responses of 20 CL reagents (three luminol derivatives, two imidazopyrazinone derivatives, eight lophine derivatives, six acridinium ester derivatives andlucigenin) against six types of ROS (superoxide anion: O-2(center dot-), hydroxyl radical: (OH)-O-center dot, hydrogen peroxide: H2O2, hypochlorite anion: ClO-, singlet oxygen: O-1(2), and nitric oxide: NO). As a result of the screening, it was found that nine CL reagents selectively detected O-2(center dot-) while one CL reagent selectively detected (OH)-O-center dot. However, no CL reagent had selectivity on the detection of H2O2, ClO-, O-1(2) and NO. Our screening results could help to select the most suitable CL reagent for selective determination of different ROS. As an application study, 4-methoxyphenyl-10-methylacridinium-9-carboxylate (MMAC), one of the acridinium ester derivatives, showed high selectivity on the detection of O-2(center dot-), and thus was applied to the assay of superoxide dismutase (SOD) activity. The dynamic range and detection limit of the developed CL assay were 0.1-10 and 0.06 U mL(-1), respectively. Significant correlation (r = 0.997) was observed between the results by the CL assay using MMAC and the spectrophotometric assay using 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt. Crown Copyright (C) 2010 Published by Elsevier B.V. All rights reserved.