Comprehensive high-throughput screening and efficient preparation of active components of Ganoderma resinaceum and in vitro evaluation of therapeutic effect on gout.

This study focused on Ganoderma resinaceum, a widely utilized substance in the fields of medicine and food. Xanthine oxidase (XOD) was employed as a biological activity indicator, while receptor-ligand UF-LC/MS (ultrafiltration affinity liquid chromatography-mass spectrometry) technology was applied to screen and identify compounds with potential anti-gout properties. Additionally, molecular docking and dynamic simulations were performed with a common gout receptor (PDB ID: 1FIQ) to investigate the mechanism of action and assess the stability of the interactions. The optimal extraction method was determined through response surface methodology, and target compounds were continuously separated using UNIFAC (universal quasichemical functional group activity coefficient-based) coupled gradient-CCC (gradient countercurrent chromatography). Nine bioactive compounds were successfully identified in G. resinaceum, with the purity of the target compounds exceeding 85.0 %. Enzymatic reaction kinetics were thoroughly analyzed to rule out any potential false positives in the experimental results. This approach is highly efficient and environmentally friendly, reducing the consumption of organic reagents while eliminating false positives. It is well-suited for high-throughput screening and separation of enzyme inhibitors in complex natural medicine systems, offering valuable insights into the application of G. resinaceum in the food and pharmaceutical industries.