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。
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 Aconi-tum pendulum Busch with semi-preparative high-speed counter-current chromatography（ HSC-CC）. 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 com-posed 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 estab-lished method is simple,highly efficient and suitable for large scale separation of ICABE from radix of Aconitum pendulum Busch.
出处
《色谱》
CAS
CSCD
北大核心
2014年第5期543-546,共4页
Chinese Journal of Chromatography
基金
中国科学院知识创新工程重要方向项目(KSCX2-EW-J-26)
关键词
半制备
高速逆流色谱
咪唑生物碱
铁棒锤
semi-preparation
high-speed counter-current chromatography （HSCCC）
imidaz-ole alkaloid
Aconitum pendulum Busch
