title: High-throughput superoxide anion radical scavenging capacity assay.
abstract: A high-throughput superoxide anion radical (O2(•-)) scavenging capacity assay based on the xanthine oxidase/xanthine reaction system was developed and validated in the present study. The reaction conditions including detection wavelength, concentrations of reactant components, reaction temperature, reaction time, pH, terminator reagent, and sample dissolving solvents were optimized. The accuracy and reliability of the assay were assessed by evaluation of linearity (r(2) = 0 .9513-0.9957), precision (intraday RSD 1.13-4.05% and interday RSD 2.13-5.62%), accuracy (95.64-97.42% recovery), and stability (RSD 2.62-6.19%), as well as comparison with the conventional colorimetric method. The EC50 values obtained by the current method and the conventional assay were highly correlated (r > 0.99). This high-throughput O2(•-) scavenging assay may be used for screening and estimating potential superoxide anion radical (O2(•-)) scavengers, especially food extracts and natural products with a very small amount of test material.
authors.affiliation: Key Laboratory of Standardization of Chinese Medicines of Ministry of Education, The Shanghai Key Laboratory for Compound Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine , Shanghai 201203, China.
authors.forename: Hongxun
authors.initials: H
authors.lastname: Tao
authors.forename: Jinge
authors.initials: J
authors.lastname: Zhou
authors.forename: Tao
authors.initials: T
authors.lastname: Wu
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Cheng
mesh_terms: D000838:Anions / Q000737:chemistry*; D016166:Free Radical Scavengers / Q000737:chemistry*; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D013481:Superoxides / Q000737:chemistry*; D014969:Xanthine Oxidase / Q000737:chemistry
chemical_list: D000838:Anions; D016166:Free Radical Scavengers; D013481:Superoxides; D014969:Xanthine Oxidase
keywords: antioxidant activity; high throughput; superoxide anion radical; xanthine oxidase