Extraction and enrichment technique of acetylcholinesterase inhibitor from Douchi

Extraction of acetylcholinesterase ( AchE) inhibitor from Douchi was optimized through single factor experiment.Crude extract was extracted by petroleum ether,methylene chloride,ethyl acetate and n butanol extract in turn,and the petroleum ether fraction was enriched with macroporous resin.Ellman’s method was used to determine AchE inhibitory activity in vitro.The results showed that the optimum extraction condition were with the extract time of 60min,ethanol concentration of 80%,extraction temperature of 70℃,ratio of liquid /solid of 1∶10. 1.0mg /mL of Douchi extract showed 52.33% AchE inhibition rate.Crude Douchi extract was further extracted.The AchE inhibition rate of 1.0mg /mL extract by following solvent was as follows: 75.52% by petroleum ether,61.00% by methylene chloride,70.84% by ethyl acetate and 26.73% by n-butanol.HZ-801 macroporous resin was selected to enrich acetylcholinesterase inhibitor.The optimum enrichment conditions were absorbed time of 3.0h,elution solvent of 80% ethanol,and 1.0mL/min elution rate.After enrichment,the AchE inhibition rate could reach 83.67%.