[关键词]
[摘要]
目的 建立高速逆流色谱分离纯化草豆蔻中山姜素和小豆蔻明的方法。方法 使用正己烷-醋酸乙酯-甲醇-水(5∶5∶7∶3)为两相溶剂系统,上相为固定相,下相为流动相,体积流量2.0 mL/min,转速800 r/min,温度25 ℃,固定相的保留率为50%,检测波长300 nm,对草豆蔻粗提物进行分离。结果 从100 mg草豆蔻粗提物中一步分离纯化得到17.2 mg山姜素和25.1 mg 小豆蔻明。经HPLC分析,质量分数分别为98.1%、99.2%,其化学结构由1H-NMR和13C-NMR鉴定。结论 与传统的、存在不可逆吸附现象的柱色谱法相比,高速逆流色谱分离纯化草豆蔻中山姜素和小豆蔻明的方法具有简单、高效、回收率高等优点。
[Key word]
[Abstract]
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.
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[基金项目]
聊城大学科研基金资助项目(X091008)