高速逆流色谱法一次性分离制备藏药蓝玉簪龙胆中的三个化合物

Separation and preparation of three compounds from Gentiana veitchiorum by high-speed counter-current chromatography(HSCCC)

目的:通过高速逆流色谱法(HSCCC)从藏药材蓝玉簪龙胆中分离制备得到三个高纯度化合物.方法:选择乙酸乙酯-正丁醇-水(4:0.5:5,v/v/v)为二相溶剂系统,上下两相分别作为固定相、流动相,主机转速设置为900 r/min,流动相溶剂流速为2 m L/min,紫外检测波长为254 nm,柱温为20℃.所采集的组分减压浓缩后烘干得到龙胆苦苷、异荭草素、异牡荆苷,应用ESI-MS/MS分析鉴定,并用HPLC测定其质量分数.结果:在260 min内可一步纯化150 mg藏药材蓝玉簪龙胆水粗提物中的龙胆苦苷、异荭草素、异牡荆苷,三者完全分离,纯度分别达到98.2%、94.5%、96.0%.结论:利用高速逆流色谱法可快速、简单、高效地富集蓝玉簪龙胆中龙胆苦苷、异荭草素、异牡荆苷三种化合物.

Objective：To separate and purify three compounds from Gentiana veitchiorum Hemsl by high-speed counter-current chromatography （HSCCC）. Methods：The solvent system of I-ISCCC was composed of ethyl acetate, u-butyl alcohol and water,in which the upper was the stationary phase,and the lower was the mobile one. The flow rate of mobile phase was 2 mL/min, and the revolution speed was set at 900 r/min with the column temperature of 20 ℃ and detection wavelength of 254 nm. The three compounds of gentiopicroside, isoorientin and isovitexin were obtained after vacuum concentration and drying, followed by deter- mination through HPLC method, and their chemical structures were identified by ESI-MS/MS method. Results：High purity gen- tiopicroside （98.2%）, i soorientin （94. 5 % ）and isovitexin （96. 0% ）were obtained from 150 mg of water crude extract through one-step separation and enrichment during 260 rain. Conclusion ： HSCCC method could be used for the preparation of three dif- ferent compounds（ gentiopicroside,isoorientin and isovitexin）from C, entiana veitchiorum Hemsl, and the method was simple, quick and efficient.