半制备型高效液相色谱法分离纯化吴茱萸中吴茱萸碱和吴茱萸次碱

Separation and Purification of Evodiamine and Rutaecarpine from Fructus Evodiae by Semi-preparative High Performance Liquid Chromatography

[目的]建立从吴茱萸中分离纯化吴茱萸碱和吴茱萸次碱的半制备型高效液相色谱法。[方法]采用C18柱(250 mm×25.4 mm I.D.,10μm),甲醇-水为流动相,考察了流动相的组成、流速和上样量对分离效果的影响,确定了适宜的色谱条件:流动相为甲醇-水(70∶30,V/V),流速为25 m L/min,上样量为60 mg,检测波长为225 nm。[结果]一次分离即可得到2种单体成分,经紫外光谱和核磁共振波谱分析得到鉴定为吴茱萸碱和吴茱萸次碱,经高效液相色谱法检测其纯度分别为99.2%和99.7%。[结论]与传统的分离方法相比,半制备型高效液相色谱法具有高效、简便、快速的特点,适用于吴茱萸中主要有效成分的快速制备。

[Objective] To establish a semi-preparative high performance liquid chromatography （SP-HPLC） method for separation and purifica-tion of evodiamine and rutaecarpine from Fructus Evodiae.[Method] A C18 column （250 mm ＆#215;25.4 mm I.D.,10 μm） was used as separation column and methanol-water as the mobile phase.The optimum separation conditions were obtained through investigating the effects caused by the composition and flow rate of the mobile phase and the sampling amount.The suitable conditions were listed as follows：methanol-water （70∶30, V/V） as mobile phase at the flow rate of 25 mL/min,the sampling amount was 60 mg,and the detection wavelength was set at 225 nm.[ Result] Under the above conditions,evodiamine and rutaecarpine were obtained through one-step separation.The purities of the two compounds were de-termined as 99.2% and 99.7% by high performance liquid chromatography （HPLC）.The chemical structures of evodiamine and rutaecarpine were identified by ultraviolet spectrum （UV） and nuclear magnetic resonance （NMR） analysis.[Conclusion] Comparing with the traditional sep-aration method,the SP-HPLC method developed in the study has many advantages such as high efficiency,facility,and high speed.And it is suit-able for fast preparation of active components from Fructus Evodiae.