Screening, Isolation, and Mass Spectrometric Analysis for Xanthine Oxidase Inhibitors from Polyporus umbellatus
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Abstract:
A new method was developed for the rapid screening of xanthine oxidase inhibitors in Polyporus umbellatus. The novel rapid and sensitive method uses an integrated approach combining a genetic algorithm-back propagation neural network mathematical model, chromatography, and mass spectrometry. Extraction of ergosterol from P. umbellatus was optimized using the genetic algorithm-back propagation neural network mathematical model. Secondly, using xanthine oxidase as the biological target molecule, ultrafiltration mass spectrometry was applied to screen for potential xanthine oxidase inhibitors in P. umbellatus. Molecular docking technology was used to validate the accuracy of the ultrafiltration experiment. Finally, with the guidance of the activity screening, high-speed counter-current chromatography was used to separate the xanthine oxidase inhibitors from P. umbellatus. The optimal extraction conditions for P. umbellatus ergosterol were 80% ethanol by volume, liquid-solid concentration of 16 mL/g, extraction time of 1.4 hours, and three extraction cycles. The optimized yield of ergosterol was 2.31%. Four xanthine oxidase inhibitors with purity >90% were identified and isolated from P. umbellatus. Their inhibitory activity ranked in order of polyporusterone A> polyporusterone B> ergosta-4,6,8(14),22-tetraen-3-one> ergosta-7,22-dien-3-one. The findings providing a theoretical basis for further studies on the anti-gout effects of P. umbellatus.
