粉防己中轮环藤酚碱的分离、纯化及鉴定

Isolation,purification and identification of cyclanoline from Stephaniae tetrandrae Radix

目的分离纯化粉防己中的轮环藤酚碱。方法采用乙醇回流提取粉防己总生物碱,利用酸溶碱沉法、正丁醇萃取法分离其水溶性生物碱;采用TLC、HPLC法跟踪鉴定生物碱;采用中性氧化铝柱层析色谱,分离水溶性生物碱,合并相同组分并重结晶;利用HPLC、质谱、核磁共振波谱对分离纯化所得轮环藤酚碱进行结构鉴定。结果粉防己中的轮环藤酚碱主要分布于正丁醇萃取部位的二氯甲烷-甲醇10∶1、10∶2洗脱部位中,HPLC纯度为98.17%;该析晶物紫外光谱存在轮环藤酚碱苯环的E、B带吸收峰,其分子离子峰m/z 342[M+H]^(+)与轮环藤酚碱分子式C_(20)H_(24)NO_(4)一致;该析晶物的^(1)HNMR及^(13)CNMR化学位移特征与文献中轮环藤酚碱结构信息一致。结论所用方法简便易行,可用于粉防己药材中轮环藤酚碱的分离纯化。

OBJECTIVE To separate and purifiyed the water-soluble alkaloid cyclanoline from Stephaniae tetrandrae Radix.METHODS The total alkaloids of S.tetrandrae Radix were extracted by ethanol reflux,and futher separated by the acid-dissolution and alkali-precipitation method and n-butanol extraction.After that,the alkaloids were tracted and identified by TLC and HPLC.The water-soluble alkaloids were further separated by nuetral alumina column chromatography,and the eluted fractions containing the same components were combined and recrystallized.The structure of cyclanoline separated from the water-soluble alkaloids was identified by HPLC,mass spectrometry and NMR spectroscopy.RESULTS Cyclanoline was mainly distributed in the n-butanol extraction part.The separation results of alumina column chromatography showed that cyclanoline was mainly distributed in the elution fractions of dichloromethane-methanol 10∶1 and 10∶2,and the purity was 98.17%by HPLC.The UV spectrum of the crystallite has E and B band absorption peaks of the benzene ring of cyclanoline,with its molecular ion peak m/z 342[M+H]^(+),consistent with the molecular formula C_(20)H_(24)NO_(4) of cyclanoline.In addition,the ^(1)HNMR and ^(13)CNMR chemical shift characteristics of the crystallite are consistent with the structure information of cyclanoline reported in the literature.CONCLUSION The method is simple and can be used for the separation and purification of cyclanoline in S.tetrandrae Radix.