夏天无生物碱的高速逆流色谱分离纯化

Separation of Alkaloids from Corydalis Decumbens (Thunb.) Pers.by High Speed Counter-Current Chromatography

采用pH-区带精制逆流色谱与常规高速逆流色谱相结合的方法快速分离纯化夏天无总生物碱。利用pH-区带精制逆流色谱对夏天无总生物碱进行分离,以正己烷-乙酸乙酯-甲醇-水(5∶5∶2∶8,V/V,上相加5mmol/L三乙胺,下相加5mmol/LHCl)为溶剂系统,上样量3.0g,分离得到1个混合物和3个高纯度的生物碱单体:原阿片碱(375mg)、苏元胡碱甲(362mg)和比枯枯灵(246mg),其纯度分别为97.5%,95.6%和97.1%。所得混合物850mg经常规高速逆流色谱二次分离,以正己烷-乙酸乙酯-甲醇-水(1∶0.8∶1.1∶1,V/V)为溶剂系统,得到比枯枯灵(105mg)和四氢巴马亭(470mg)单体,纯度分别为99.1%和99.7%。从该药材分得苏元胡碱甲,两种制备分离模式相结合,提高了夏天无生物碱的分离效率。

A method was established for the separation and purification of alkaloids from the crude extract of Corydalis decumbens（Thunb.） Pers. using high speed counter-current chromatography（HSCCC） with two modes. First, pH-zone-refining counter-current chromatography was successfully performed with a two-phase solvent system, n-hexane-ethyl acetate-methanol-water（5∶5∶2∶8, V/V）, in which 5 mmol/L triethylamine in the organic stationary phase and 5 mmol/L hydrochloric acid in the aqueous mobile phase. From 3.0 g of crude extract, a mixture and three compounds, with 375 mg protopine（1）, 362 mg humosine A（2） and 246 mg bicuculline（3） were obtained with purities of 97.5%, 95.6% and 97.1%, respectively. And then, the mixture was separated by HSCCC with the two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water（1∶0.8∶1.1∶1, V/V）. Bicuculline（105 mg） and tetrahydropalmatine（470 mg） were obtained from 850 mg mixture. Among them, humosine A was isolated from Corydalis decumbens（Thunb.） Pers. for the first time. pH-Zone refining counter-current chromatography combined with routine HSCCC improved the separation efficiency of alkaloids from crude Corydalis decumbens（Thunb.） Pers.