薄层色谱图像分析定量-高速逆流色谱法快速分离制备吴茱萸中的三个生物碱类化合物  

Separation and purification of three alkaloids from Evodia rutaecarpa( Juss. ) benth by high-speed counter-current chromatography and their quantification by thin layer chromatography using image analysis

作　　者：韦玮[1] 周盈利[1] 唐铁鑫[1] 李雯婷[1] 杨得坡[1,2] 朱龙平[1,2] 王冬梅[1] 

机构地区：[1]中山大学药学院天然药物与中药研究所,广州510006 [2]广东省现代中药工程技术研究开发中心,广州510006

出　　处：《分析试验室》2014年第1期109-112,共4页Chinese Journal of Analysis Laboratory

摘　　要：建立了薄层色谱图像分析定量-高速逆流色谱法从吴茱萸甲醇提取物中快速分离制备吴茱萸碱、吴茱萸次碱、吴茱萸卡品碱3个化合物的方法。采用数码相机对HSCCC溶剂系统上下相中的目标化合物进行TLC图像采集,使用积分软件OPSIA计算各溶剂系统对应的K值,筛选出最佳溶剂系统正己烷-乙酸乙酯-甲醇-水(3:2.5:3.5:2,V/V)。取上相为固定相,下相为流动相,检测波长为254 nm,在主机转速860 r/min、流动相流速1.5 mL/min的条件下,一次性从300 mg样品中分离制备得到经1H-NMR和13C-NMR确证的吴茱萸碱(15.3mg)、吴茱萸次碱(10.1 mg)、吴茱萸卡品碱(20.7 mg),纯度分别为93.6%,97.3%和95.1%(HPLC法)。A method by high-speed counter-current chromatography （HSCCC） combined with thin layer chroma- tography quantification using image analysis （TLCQIA） method was established to isolate and determine three al- kaloids from the methanol extract of Evodia rutaecarpa （ Juss. ） Benth （ Wuzhuyu in Chinese） with a two phase solvent system composed of n-hexane-ethyl acetate-methanol-water （3：2. 5：3.5：2, V/V）. The developed solvent system for TLC analysis was optimized and the digital images of targeted compounds on the TLC plate at 254 nm were acquired by a digital camera. Quantitative analysis of the targeted compounds in the upper and lower phases was conducted with an OPSIA software in computer and the K values of solvent systems could be obtained from the ratio of integral area. The HSCCC conditions were selected as follows： the upper phase was used as the sta- tionary phase and the lower phase as the mobile phase, the flow rate of the mobile phase was 1.5 mL/min, theapparatus rotated at 860 r/min, and the detection wavelength was 254 nm. Under these optimized conditions, evodiamine （ 15.3 mg）, rutaecarpine （ 10. 1 mg） and evocarprine （20. 7 rag） were isolated efficiently from 300 mg of the methanolic extract of E. rutaecarpa by onetime HSCCC separation with purities of 93.6%, 97.3%, 95. 1%, respectively, and their structures were confirmed by NMR. The TLCQIA-HSCCC method established was proved to be as effective as the HPLC-I-ISCCC method but with the advantages of more convenient, short time-consuming and low costs.

关 键 词：薄层色谱图像分析定量法 高速逆流色谱法 吴茱萸 生物碱 

分 类 号：O657.7[理学—分析化学]