title: Screening of Potential Xanthine Oxidase Inhibitors in Gnaphalium hypoleucum DC. by Immobilized Metal Affinity Chromatography and Ultrafiltration-Ultra Performance Liquid Chromatography-Mass Spectrometry.
abstract: In this study, a new method based on immobilized metal affinity chromatography (IMAC) combined with ultrafiltration-ultra performance liquid chromatography-mass spectrometry (UF-UPLC-MS) was developed for discovering ligands for xanthine oxidase (XO) in Gnaphalium hypoleucum DC., a folk medicine used in China for the treatment of gout. By IMAC, the high flavonoid content of G. hypoleucum could be determined rapidly and efficiently. UF-UPLC-MS was used to select the bound xanthine oxidase ligands in the mixture and identify them. Finally, two flavonoids, luteolin-4'-O-glucoside and luteolin, were successfully screened and identified as the candidate XO inhibitors of G. hypoleucum. They were evaluated in vitro for XO inhibitory activity and their interaction mechanism was studied coupled with molecular simulations. The results were in favor of the hypothesis that the flavonoids of G. hypoleucum might be the active content for gout treatment by inhibiting XO.
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. jian871211@sina.com.
authors.forename: Hong-Jian
authors.initials: HJ
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. huyijuan604@163.com.
authors.forename: Yi-Juan
authors.initials: YJ
authors.lastname: Hu
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. xpan840520@163.com.
authors.forename: Pan
authors.initials: P
authors.lastname: Xu
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. jxlwq22@163.com.
authors.forename: Wei-Qing
authors.initials: WQ
authors.lastname: Liang
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. Kenzei@163.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. lpgzaas@163.com.
authors.forename: Pei-Gang
authors.initials: PG
authors.lastname: Liu
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. Lemon2193@163.com.
authors.forename: Lin
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, 132 Tianmushan Road, Hangzhou 310007, China. pjb0225@163.com.
authors.forename: Jin-Bao
authors.initials: JB
authors.lastname: Pu
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000302:isolation & purification*; D005419:Flavonoids / Q000737:chemistry* / Q000302:isolation & purification*; D036622:Gnaphalium / Q000737:chemistry*; D013058:Mass Spectrometry; D014462:Ultrafiltration; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors* / Q000737:chemistry
chemical_list: D004791:Enzyme Inhibitors; D005419:Flavonoids; D014969:Xanthine Oxidase
keywords: Gnaphalium hypoleucum DC.; XO inhibitor; immobilized metal affinity chromatography; luteolin; luteolin-4′-O-glucoside