title: Ultrahigh performance liquid chromatography-triple quadrupole mass spectrometry inhibitors fishing assay: a novel method for simultaneously screening of xanthine oxidase inhibitor and superoxide anion scavenger in a single analysis.
abstract: Xanthine oxidase (XOD) inhibitors and superoxide anion scavengers play an important role in the treatment of gout and the inhibition of many diseases related to superoxide anion. The respective quantitation of uric acid and superoxide anion by traditional spectroscopic methods is routine in XOD inhibitors and superoxide anion scavengers screening at laboratories worldwide. In the present study, we established an ultrahigh performance liquid chromatography and triple quadrupole mass spectrometry (UHPLC-TQ-MS) method of higher accuracy and speed that combines screening of superoxide anion scavenger and XOD inhibitor in a single analysis by adding WST-1 (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium sodium salt) to the enzymatic reaction. We applied the established method to determine the XOD inhibitory activities and superoxide scavenging activities of some herbal extracts and compounds from natural products, which could be classified into six groups based on the results of the assay. Our innovative protocol is fast, accurate and robust. Moreover, it can eliminate false positive and false negative results which may occur in the traditional spectroscopic methods.
authors.affiliation: Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun Center of Mass Spectrometry, Changchun, PR China.
authors.forename: Shu
authors.initials: S
authors.lastname: Liu
authors.forename: Junpeng
authors.initials: J
authors.lastname: Xing
authors.forename: Zhong
authors.initials: Z
authors.lastname: Zheng
authors.forename: Fengrui
authors.initials: F
authors.lastname: Song
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Liu
authors.forename: Shuying
authors.initials: S
authors.lastname: Liu
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000494:pharmacology; D005562:Formazans / Q000032:analysis; D016166:Free Radical Scavengers / Q000032:analysis* / Q000494:pharmacology; D057230:Limit of Detection; D013481:Superoxides / Q000037:antagonists & inhibitors*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013778:Tetrazolium Salts / Q000032:analysis; D014527:Uric Acid / Q000032:analysis; D019820:Xanthine / Q000032:analysis; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors*
chemical_list: C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D004791:Enzyme Inhibitors; D005562:Formazans; D016166:Free Radical Scavengers; D013778:Tetrazolium Salts; D013481:Superoxides; D019820:Xanthine; D014527:Uric Acid; D014969:Xanthine Oxidase