title: Selective fishing and analysis of xanthine oxidase binders from two Fabaceae species by coupling enzyme functionalized core-shell magnetic nanoparticles with HPLC-MS.
abstract: Xanthine oxidase (XOD) immobilized core-shell magnetic silica (Fe3O4@SiO2-XOD) nanoparticles coupled with high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed to fish out and analyze XOD binders from two Fabaceae species, Puerariae lobata flower and Glycyrrhiza uralensis root. The prepared Fe3O4@SiO2-XOD nanoparticles exhibited good specificity for XOD binders, better dispersion in aqueous solution and reusability than those of Fe3O4-XOD nanoparticles. The amount of XOD immobilized onto Fe3O4@SiO2 nanoparticles was 339.9μg/mg and the activity of Fe3O4@SiO2-XOD nanoparticles remained 95% after ten times usage. The optimum conditions of selective fishing were optimized, and finally incubating pH was set at 7, incubating temperature at 25°C and adsorption time at 30min. Twelve XOD binders were successfully identified from ethyl acetate extract of P. lobata flower and G. uralensis root. The developed method provides a rapid, purposeful and effective way to identify active compounds from natural complex mixtures.
authors.affiliation: School of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China; Key Laboratory of Resources Chemistry of Nonferrous Metals, Central South University, Changsha 410083, China.
authors.forename: Liangliang
authors.initials: L
authors.lastname: Liu
authors.affiliation: School of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Shuyun
authors.initials: S
authors.lastname: Shi
authors.affiliation: School of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Huading
authors.initials: H
authors.lastname: Zhao
authors.affiliation: School of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Jingang
authors.initials: J
authors.lastname: Yu
authors.affiliation: School of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.
authors.forename: Xinyu
authors.initials: X
authors.lastname: Jiang
authors.affiliation: School of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China; Key Laboratory of Resources Chemistry of Nonferrous Metals, Central South University, Changsha 410083, China. Electronic address: xqchen@csu.edu.cn.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Chen
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D055808:Drug Discovery; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D007887:Fabaceae / Q000737:chemistry*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D008892:Milk / Q000201:enzymology; D010936:Plant Extracts / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D014969:Xanthine Oxidase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D010936:Plant Extracts; D014969:Xanthine Oxidase
keywords: Glycyrrhiza uralensis root; HPLC–MS; Magnetic nanoparticles; Puerariae lobata flower; Selective fishing; Xanthine oxidase