Simple and rapid chemiluminescence (CL) assays for H2O2, •OH, •O2−and1O2using 5-amino-2,3-dihydro-1,4-phthalazinedione (luminol) or 2-methyl-6-phenyl-3,7-dihydroimidazo[1,2-α]pyrazin-3-one (CLA) as CL reagents were developed. The means of the intra-assay relative standard deviations of ten replicate measurements of H2O2(25-120 μM), •OH generated from Fe(II) ion (2.5-10 μM) in the presence of 980 μM H2O2, •O2−generated from hypoxanthine (HX) (7-50 μM) in the presence of 9 × 10−3units xanthine oxidase (XO) and1O2generated from NaOCl (3-12 mM) in the presence of 97.6 μM H2O2were found to be 4.0%, 2.8%, 2.4% and 8.7%, respectively. To validate the proposed methods, the scavenging abilities of three standard antioxidative compounds, such as L-ascorbic acid, (±)-α-tocopherol and superoxide dismutase (SOD) were examined for four active oxygen species and compared with those by anelectron spin resonance (ESR) spin-trapping method. In addition, the CL methods were also applied to establish the relationships between the decrease of CL intensity and the structures as well as redox characters of syringic acid, 3-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid. From the obtained results, the scavenging effects to H2O2, •OH, •O2−and1O2of other dihydroxybenzoic acids were also evaluated.