title: Screening inhibitors of xanthine oxidase from natural products using enzyme immobilized magnetic beads by high-performance liquid chromatography coupled with tandem mass spectrometry.
abstract: In this study, high-performance liquid chromatography coupled with tandem mass spectrometry was used to assess the results of bioactive compound screening from natural products using immobilized enzyme magnetic beads. We compared three commercial magnetic beads with modified amino, carboxy, and N-hydroxysuccinimide groups, respectively. Amino magnetic beads performed best for immobilization and were selected for further experiments. Xanthine oxidase was immobilized on amino magnetic beads and applied to screen potential inhibitors in fresh Zingiber officinale Roscoe, extracts of Scutellaria baicalensis Georgi, and Pueraria lobata Ohwi. In total, 12 potential xanthine oxidase ligands were identified from fresh Zingiber root and Scutellaria root extracts, of which eight were characterized and the concentration required for 50% inhibition was determined. Preliminary structure-function relationships were discussed based on these results. A convenient and effective method was therefore developed for the identification of active compounds from complex natural product mixtures.
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, China Pharmaceutical University, Nanjing, P.R. China.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, China Pharmaceutical University, Nanjing, P.R. China.
authors.forename: Dapeng
authors.initials: D
authors.lastname: Li
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, China Pharmaceutical University, Nanjing, P.R. China.|State Key Laboratory of Natural Medicines, China Pharmaceutical University, Jiangsu, Nanjing, P.R. China.
authors.forename: Boyang
authors.identifier: http://orcid.org/0000-0002-1401-9250
authors.initials: B
authors.lastname: Yu
authors.affiliation: Jiangsu Key Laboratory of TCM Evaluation and Translational Research, China Pharmaceutical University, Nanjing, P.R. China.|State Key Laboratory of Natural Medicines, China Pharmaceutical University, Jiangsu, Nanjing, P.R. China.
authors.forename: Jin
authors.initials: J
authors.lastname: Qi
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004791:Enzyme Inhibitors / Q000302:isolation & purification*; D010936:Plant Extracts / Q000737:chemistry; D029908:Pueraria / Q000737:chemistry*; D031364:Scutellaria baicalensis / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors*; D030521:Zingiberaceae / Q000737:chemistry*
chemical_list: D004791:Enzyme Inhibitors; D010936:Plant Extracts; D014969:Xanthine Oxidase
keywords: inhibitors; magnetic beads; natural products; xanthine oxidase