目的 建立高速逆流色谱(HSCCC)法从裂叶独活根中分离制备高质量分数的香豆素类成分。方法 利用高效液相色谱-二极管阵列检测器分析优化溶剂体系,选择正己烷-醋酸乙酯-甲醇-水(1.5:2.5:2:1.5)为HSCCC溶剂系统,上相为固定相,下相为流动相,主机转速为850 r/min,体积流量为3 mL/min,检测波长为323 nm。所接收馏份在50 ℃减压浓缩,静置得到蛇床子素晶体、二氢欧山芹醇乙酸酯晶体和二氢欧山芹醇当归酸酯晶体,应用核磁共振氢谱、碳谱、质谱、红外光谱等进行鉴定,并用HPLC测定质量分数。结果 进样量为300 mg浸膏,得到二氢欧山芹醇乙酸酯6.3 mg、蛇床子素10.6 mg、二氢欧山芹醇当归酸酯6.4 mg,三者的质量分数均达到96.0%以上。结论 HSCCC法可用于裂叶独活中高质量分数香豆素类成分的快速分离制备。

Objective To separate and purify coumarins from Heracleum millefolium by high-speed counter-current chromatography (HSCCC). Methods HPLC was used to analyze and optimize the solvent system and hexane-ethyl acetate-water-methanol (1.5:2.5:2:1.5) was chosen as the two-phase solvent system of HSCCC, in which the upper phase was used as the stationary phase, while the lower phase was used as the mobile phase with flow rate of 3 mL/min, the revolution speed was set at 850 r/min, and detected at 323 nm. Three compounds were obtained as crystals by vacuum concentration at 50 ℃, and determined through HPLC, then their structures were identified by IR, ESI-MS, ¹H-NMR, and ¹³C-NMR. Results Columbianetin acetate (6.3 mg), osthole (10.6 mg), and columbianadin (6.4 mg) were obtained from crude sample (300 mg) extracts and their purity were above 96.0%. Conclusion HSCCC is a powerful technique for the rapid separation and purification of coumarins from crude extract of H. millefolium.

江西省科技支撑计划(20112BBG70021);江西中医药大学道地药材及特色中药资源研究专项;西藏自治区自然科学基金资助项目(2015ZR-14-65)