目的 建立高速逆流色谱（HSCCC）技术高效快速地分离制备益母草中益母草碱的方法。方法 单因素实验优化益母草碱的提取工艺，而后考察不同的HSCCC溶剂体系，根据分配系数（K），确定溶剂体系为醋酸乙酯-正丁醇-水（3：2：5），以其上相为固定相，下相为流动相，转速850 r/min，体积流量2.2 mL/min，检测波长为277 nm，对益母草样品进行HSCCC分离。结果 益母草正丁醇萃取部位通过HSCCC可分离得到益母草碱，该方法成功应用于益母草粗提物中益母草碱的分离：1次HSCCC运行可从2.48 g益母草70%乙醇粗提物（益母草碱质量分数为3.01%）中分离得到68 mg益母草碱，质量分数为96.2%，得率为2.74%。结论 建立的方法可高效分离纯化益母草中的益母草碱，为得到高纯度的益母草碱提供了制备技术。

Objective To establish a simple and effective method for the isolation of leonurine from Leonurus japonicus by high speed countercurrent chromatography (HSCCC). Methods The extraction conditions of leonurine were optimized by one-factor experimental design. After comparing several different solvent systems, the two phase system of ethyl acetate-n-butanol-water (3:2:5) was finally chosen as operating solvent of HSCCC for the separation of leonurine, in which the lower phase was determined as the mobile phase and the upper phase as stationary. The detection of eluates was performed with an ultraviolet detector at 277 nm. The rotation speed was adjusted at 850 r/min, and the flow rate was 2.2 mL/min. Results Leonurine was successively isolated from n-butanol fraction by HSCCC, and the above established method was also successively applied to the crude extact of L.japonicus. Finally, 68 mg of leonurine with purity about 96.2% could be obtained from 2.48 g crude extract of L.japonicus in a single injection. Conclusion The described approaches actively promote efficient preparation strategy to obtain high purity of leonurine.

广东省省部产学研合作专项资金项目（2013B090500100）