pH区带逆流色谱法分离纯化吴茱萸中的吴茱萸碱和去氢吴茱萸碱

Separation and Purification of Evodiamine and Dehydroevodiamine from Evodia rutaecarpa by pH Zone Countercurrent Chromatography

目的分离纯化吴茱萸中的活性成分。方法应用p H区带逆流色谱,以氯仿-乙酸乙酯-甲醇-水(3:1:3:3,v/v)为两相溶剂体系,上相加入10 mmol/L盐酸作为固定相;下相加入5 mmol/L三乙胺作为流动相,转速850 r/min,流速2.0 m L/min,检测波长254 nm,所得馏分经HPLC检测,并经电喷雾电离(ESI)质谱和核磁共振谱(NMR)鉴定化合物的结构。结果从2 g吴茱萸粗提物中一次性分离得101 mg吴茱萸碱和304 mg去氢吴茱萸碱,其纯度为90.0%和93.1%。结论此法简便、快捷、重现性好、上样量大,适于制备性分离吴茱萸碱和去氢吴茱萸碱。

Objective To separate and purify the active ingredients from Evodia rutaecarpa. Methods A pH zone countercurrent chromatography was used with chloroform-ethyl acetate-methanol-water （3：1：3：3, v/v） as a two- phase solvent system; the upper phase was used as the stationary phase by adding 10 mmol/L hydrochloric acid, the lower phase was used as the mobile phase by adding 5 mmol/L triethylamine, with the rotary speed of 850 r/min and a flow rate of 2.0 mL/min. The detection wavelength was 254 nm. The separated fractions were detected by HPLC, and the structures of the compounds were identified by ESI-MS and NMR. Results Evodiamine （101 mg） and dehydroevodiamine （304 mg） were isolated from 2 g crude extract of Evodia rutaecarpa, with the purity of 90.0 % and 93.1%, respectively. Conclusion The method is simple, rapid, reproducible and of large loading capacity, and suitable for the preparative separation of evodiamine and dehydroevodiamine.