色谱

色谱 ›› 2014, Vol. 32 ›› Issue (5): 543-546.DOI: 10.3724/SP.J.1123.2013.12007

• 技术与应用 • 上一篇    下一篇

半制备型高速逆流色谱分离制备铁棒锤根中的一种咪唑生物碱

刘永玲1,2, 陈涛1, 陈晨1, 邹登朗1,2, 李玉林1   

  1. 1. 中国科学院西北高原生物研究所, 青海 西宁 810008;
    2. 中国科学院大学, 北京 100049
  • 收稿日期:2013-12-10 修回日期:2014-01-23 出版日期:2014-05-08 发布日期:2014-04-26
  • 通讯作者: 李玉林
  • 基金资助:

    中国科学院知识创新工程重要方向项目（KSCX2-EW-J-26）.

Isolation and preparation of an imidazole alkaloid from radix of Aconitum pendulum Busch by semi-preparative high-speed counter-current chromatography

LIU Yongling1,2, CHEN Tao1, CHEN Chen1, ZOU Denglang1,2, LI Yulin1   

  1. 1. Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, China;
    2. University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2013-12-10 Revised:2014-01-23 Online:2014-05-08 Published:2014-04-26

PDF

302

被引次数

13

摘要：

采用高速逆流色谱（HSCCC）技术从铁棒锤根氯仿提取物中分离制备了一种高纯度咪唑类生物碱1H-imidazole-2-carboxylic acid，butyl ester （ICABE）。采用高效液相色谱（HPLC）测定目标化合物在两相溶剂中的分配系数，优化HSCCC分离ICABE的溶剂体系，确定了以正己烷-氯仿-乙醇-水（10：1：13：2，v/v/v/v）为HSCCC的两相溶剂系统，以上相为固定相，下相为流动相，流动相流速为1.8 mL/min，主机转速850 r/min，检测波长为230 nm条件下进行分离制备，在350 min内从100 mg粗样品中一步分离得到7.5 mg ICABE，经HPLC检测其纯度达98%以上（峰面积归一化法），结构由UV、1H-NMR和13C-NMR得以鉴定。该方法简便、快速，所得产物纯度高，适合于铁棒锤中ICABE的制备分离。

关键词: 半制备, 高速逆流色谱, 咪唑生物碱, 铁棒锤

Abstract:

Aconitum pendulum Busch is rich C19 diterpenoid alkaloids, but there is no report of imidazole alkaloid in Aconitum pendulum Busch. In this study, an imidazole alkaloid named 1H-imidazole-2-carboxylic acid, butyl ester (ICABE) was successfully separated from Aconitum pendulum Busch with semi-preparative high-speed counter-current chromatography (HSCCC). The partition coefficient was measured by HPLC to select the solvent systems for ICABE separation by HSCCC. The separation was performed with a two-phase solvent system composed of n-hexane-chloroform-ethanol-water (10:1:13:2, v/v/v/v). The upper phase was used as the stationary phase and the lower phase as the mobile phase. It was operated at a flow rate of 1.8 mL/min. The apparatus was rotated at 850 r/min, and the detection wavelength was set at 230 nm. Under the selected conditions, a high efficiency separation of HSCCC was achieved, and 7.5 mg of ICABE was obtained from 100 mg of the crude sample of Aconitum pendulum in one-step separation within 350 min. The HPLC analysis showed that the purity of the compound was over 98%. The chemical structure was confirmed by UV, 1H-NMR and 13C-NMR. The established method is simple, highly efficient and suitable for large scale separation of ICABE from radix of Aconitum pendulum Busch.

Key words: Aconitum pendulum Busch, high-speed counter-current chromatography (HSCCC), imidazole alkaloid, semi-preparation

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