Efficient combination of ionic-liquid-based ultrasound-assisted extraction, complex chromatography, and molecular docking for screening of acetylcholinesterase inhibitors from Ganoderma atrum

Compounds extracted from Ganoderma atrum ( G. atrum ) is significantly beneficial to human health, have been incorporated in numerous remedies owing to their brain and kidney health benefits and antiinflammatory and diuretic properties. G. atrum a pilot of homology of medicine and food, has an important development value of functional food and health food. In this study, to develop a new ionic-liquid-based ultrasound-assisted extraction method and bio-affinity ultrafiltration coupled with LC and MS. Moreover, molecular docking was used to evaluate the activity of the AChE inhibitors, which were subsequently isolated and purified using consecutive counter-current chromatography (CCC) and semi-preparative high-performance liquid chromatography (HPLC). Lastly, the active compounds were identified using MS and nuclear magnetic resonance. Four AChE inhibitors, viz . ergosterol, ergosta-7,22-dien-3-one, ergosta-7-en-3 β -ol, and ergosta-4,6,8(14),22-tetraen-3-one with purities of 92.23%, 90.19%, 90.52% and 96.72%, respectively, were obtained from G. atrum using consecutive CCC (ethyl acetate/methanol/water, 4.0:1.0:7.0, v/v/v ) coupled with semi-preparative HPLC (4% A (0–30 min), 4–100% A (30–40 min)). Our results suggested that G. atrum extracts can be used in the potential prevention and treatment of neurodegenerative diseases. It will help to elucidate the mechanisms of action of various medicinal plants at the molecular level and provide more opportunities to discover and develop new potential therapeutic remedies from health care food resources.