To identify compounds derived from kudzu root (Pueraria lobata) with inhibitory activity against xanthine oxidase (XOD), this study first prepared core-shell magnetic silica nanoparticles immobilized with XOD (Fe3O4@SiO2-XOD).By integrating high-performance liquid chromatography mass spectrometry and molecular docking simulations, compounds isolated from the screened kudzu root extract were further separated, and their binding characteristics with XOD were analyzed.Results indicated that the synthesized Fe3O4@SiO2 XOD nanoparticles exhibited excellent specificity for XOD-binding ligands and proved highly effective for ligand screening applications.A total of 15 compounds were successfully identified from the kudzu root extract, including 9 isoflavones, 3 phenolic acids, 2 phytosterols, and 1 fatty acid.Molecular docking simulations were performed involving xanthine, hypoxanthine, allopurinol, and the screened compounds against XOD.Results demonstrated that all 15 compounds exhibited stronger binding affinity (lower binding energy) to XOD compared to the substrates (xanthine and hypoxanthine) and the reference inhibitor allopurinol.Among these, isoflavones showed the highest potency, forming the first tier of inhibitors.Phytosterols, which primarily rely on hydrophobic interactions, constituted the second tier.Phenolic acids and fatty acids comprised the third tier.All 15 compounds were identified as effective XOD inhibitors.This study establishes an efficient and targeted methodology for the rapid identification of bioactive compounds from complex natural mixtures.
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