title: Quick identification of xanthine oxidase inhibitor and antioxidant from Erycibe obtusifolia by a drug discovery platform composed of multiple mass spectrometric platforms and thin-layer chromatography bioautography.
abstract: As a final step of the purine metabolism process, xanthine oxidase catalyzes the oxidation of hypoxanthine and xanthine into uric acid. Our research has demonstrated that Erycibe obtusifolia has xanthine oxidase inhibitory properties. The purpose of this paper is to describe a new strategy based on a combination of multiple mass spectrometric platforms and thin-layer chromatography bioautography for effectively screening the xanthine oxidase inhibitory and antioxidant properties of E. obtusifolia. This strategy was accomplished through the following steps. (i) Separate the extract of E. obtusifolia into fractions by an autopurification system controlled by liquid chromatography with mass spectrometry. (ii) Determine the active fractions of E. obtusifolia by thin-layer chromatography bioautography. (iii) Identify the structure of the main active compounds with the information provided by direct analysis in real time mass spectrometry. (iv) Calculate the IC50 value of each compound against xanthine oxidase using high-performance liquid chromatography. Using the caulis of E. obtusifolia as the experimental material, seven target peaks were screened out as xanthine oxidase inhibitors or antioxidants. Our screening strategy allows for rapid analysis of small molecules with almost no sample preparation and can be completed within a week, making it a useful assay to identify unstable compounds and provide the empirical foundation for E. obtusifolia as a natural remedy for gout and oxidative-stress-related diseases.
authors.affiliation: The MOE Key Laboratory for Standardization of Chinese Medicines and The Shanghai Key Laboratory for Compound Chinese medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, P. R. China; Department of Pharmacognosy, China Pharmaceutical University, Nanjing, P. R. China.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Chen
authors.forename: Hongxun
authors.initials: H
authors.lastname: Tao
authors.forename: Liping
authors.initials: L
authors.lastname: Liao
authors.forename: Zijia
authors.initials: Z
authors.lastname: Zhang
authors.forename: Zhengtao
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000975:Antioxidants / Q000737:chemistry*; D001331:Automation; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D002855:Chromatography, Thin Layer; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006073:Gout / Q000188:drug therapy; D020128:Inhibitory Concentration 50; D013058:Mass Spectrometry; D018384:Oxidative Stress; D010936:Plant Extracts / Q000737:chemistry; D014326:Tropanes / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors*
chemical_list: D000975:Antioxidants; D004791:Enzyme Inhibitors; D010936:Plant Extracts; D014326:Tropanes; C027863:bao gong teng A; D014969:Xanthine Oxidase
keywords: Direct analysis in real time; Erycibe obtusifolia; Thin-layer chromatography bioautography; Xanthine oxidase inhibitors