Efficient and fast screening and separation based on computer‐aided screening and complex chromatography methods for lipoxygenase inhibitors from Ganoderma lucidum

Abstract Introduction Accurate screening and targeted preparative isolation of active substances from natural medicines have long been technical challenges in natural medicine research. Objectives This study outlines a new approach for improving the efficiency of natural product preparation, focusing on the rapid and accurate screening of potential active ingredients in Ganoderma lucidum and efficient preparation of lipoxidase inhibitors, with the aim of providing new ideas for the treatment of Alzheimer's disease with G. lucidum . Methods The medicinal plant G. lucidum was selected through ultrafiltration coupled with liquid chromatography and mass spectrometry (UF–LC–MS) and computer‐assisted screening for lipoxygenase (LOX) inhibitors. In addition, the inhibitory effect of the active compounds on LOX was studied using enzymatic reaction kinetics, and the underlying mechanism is discussed. Finally, based on the earlier activity screening guidelines, the identified ligands were isolated and purified through complex chromatography (high‐speed countercurrent chromatography and semi‐preparative high‐performance liquid chromatography). Results Five active ingredients, ganoderic acids A, B, C 2 , D 2 , and F, were identified and isolated from G. lucidum . We improved the efficiency and purity of active compound preparation using virtual computer screening and enzyme inhibition assays combined with complex chromatography. Conclusion The innovative methods of UF–LC–MS, computer‐aided screening, and complex chromatography provide powerful tools for screening and separating LOX inhibitors from complex matrices and provide a favourable platform for the large‐scale production of bioactive substances and nutrients.