目的 建立对荷叶生物碱进行分离的pH区带逆流色谱方法。方法 溶剂系统为叔丁基甲醚-甲醇-水(4∶1∶5),上相添加三乙胺(10 mmol/L)作为固定相,下相添加盐酸(5 mmol/L)为流动相进行洗脱,在主机转速800 r/min、体积流量1.5 mL/min、检测波长254 nm条件下进行分离制备。结果 从2.18 g荷叶提取物中一次分离得到N-去甲荷叶碱110 mg、O-去甲荷叶碱420 mg、荷叶碱310 mg和莲碱190 mg 4种生物碱,质量分数均大于98%;各化合物通过HPLC、MS和1H-NMR进行了化学结构鉴定。结论 pH区带逆流色谱法是一种快速高效的分离纯化荷叶生物碱的方法。

Objective A novel method for the separation of alkaloids from Nelumbinis Folium by pH-zone-refining counter-current chromatography (pH-ZRCCC). Methods The two-phase solvent system composed of MtBE-methanol-water (4:1:5), 10 mmol/L triethylamine (TEA) in organic stationary phase and 5 mmol/L HCl in aqueous mobile phase. The upper phase was used as the stationary phase, while the lower phase was used as the mobile phase. In one-step elution under the conditions of a flow rate of 1.5 mL/min, 800 r/min and the effluent was detected at 254 nm. Results From 2.18 g of the crude extract, 110 mg N-nornuciferin, 420 mg O-nornuciferin, 310 mg nuciferine, and 190 mg roemerine were obtained and each with a purity of over 98% as determined by HPLC. The structures of the isolated Compounds were identified by HPLC, MS, and 1H-NMR. Conclusion The present pH-ZRCCC method may be applied to the purification of other various alkaloids from natural products.

国家自然科学基金资助项目(20872083);中国中医科学院中药研究所自然基金资助项目(ZZ20090107);山东省科技攻关项目(2010GSF10287);济南市高等院所自主创新计划项目(201004010)