高速逆流色谱分离纯化钩吻中钩吻素甲

Separation and Purification of Gelsemine from Gelsemium Elegans by High-speed Counter-current Chromatography

采用高速逆流色谱法,分别以正己烷-乙酸乙酯-无水乙醇-水(3∶3∶2∶3 V/V)和氯仿-甲醇-0.2 mol/L盐酸(4∶3∶1.5 V/V)为溶剂体系,从300 mg钩吻总碱中分离纯化出一种钩吻生物碱单体30.78 mg,高效液相色谱技术分析其质量分数为97.76%,核磁共振谱、质谱分析确证其为钩吻素甲;通过小鼠醋酸扭体法检测,表明钩吻总碱和钩吻素甲对小鼠均具有显著的镇痛活性。高速逆流色谱技术可高效分离纯化具有镇痛活性的钩吻素甲。

30.78 mg of gelsemine was separated from 300 mg of crude extract of Gelsemium elegans by high-speed count- er-current chromatography （HSCCC） twice with the two-phase solvent system composed of hexane-ethyl acetate-ethanol- water （3： 3： 2：3 in volume ratio） and chloroform-methanol-O. 2 mol/L hydrochloric acid （4： 3：1.5 in volume ratio） re- spectively. The purity of the compound was 97.76% based on high performance liquid chromatography （HPLC） analy- sis. The chemical identity of the compound was confirmed by ESI-MS,＇ H NMR and 13 C NMR. Both the total alkaloids and the isolated gelsemine had obvious analgesic effects in the acetic acid writhing tests on mice. The result indicated that the HSCCC method developed in this study separated and purified gelsemine high-efficiently.