夏天无生物碱的高速逆流色谱分离纯_临床医学论文.docVIP

夏天无生物碱的高速逆流色谱分离纯_临床医学论文 夏天无生物碱的高速逆流色谱分离纯_临床医学论文 作者：王岱杰 刘建华 耿岩玲 王晓 段文娟 傅茂润 【摘要】 　　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5 mmol/L三乙胺，下相加5 mmol/L HCl)为溶剂系统，上样量3.0 g，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(375 mg)、苏元胡碱甲(362 mg)和比枯枯灵(246 mg)， 其纯度分别为97.5%，95.6%和97.1%。所得混合物850 mg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(105 mg)和四氢巴马亭(470 mg)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。 【关键词】 夏天无; pH区带精制逆流色谱; 高速逆流色谱; 生物碱 　　Abstract A method was established for the separation and purification of alkaloids from the crude extract of Corydalis decumbens(Thunb.) Pers. using high speed counterrmed 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. 　　Keywords Corydalis decumbens(Thunb.) Pers.; pHZone refining countercurrent chromatography; High speed countercurrent chromatography; Alkaloids 　　1 引 言 　　夏天无为罂粟科植物伏生紫堇 (Corydalis decumbens (Thunb.) Pers.) 的干燥块茎，具有活血通络，行气止痛的功效，主要用于治疗中风偏瘫、跌扑损伤、风湿性关节炎、坐骨神经痛等[1]。夏天无的主要活性成分为生物碱类化合物，如原阿片碱、比枯枯灵和四氢巴马亭等[2,3]。建立主要活性成分的分离制备方法，对夏天无药材及其产品的质量控制十分必要。传统的柱层析方法存在耗时长、容易造成样品的不可逆吸附、变性及样品回收率低等缺点。高速逆流色谱(HSCCC)是一种不用固态载体的液液分配色谱，它根据样品在两相溶剂中分配