高速逆流色谱分离制备甜瓜蒂中葫芦素类化合物

Preparative Isolation and Purification of Cucurbitacins from Pedicellus Melo by High-Speed Counter-corrent Chromatography

采用高速逆流色谱法分离制备了甜瓜蒂中3种葫芦素类化合物。以正己烷-乙酸乙酯-甲醇-水(1.2∶0.8∶0.8∶1.2,V/V)为两相溶剂系统,上相为固定相,下相为流动相。在主机转速800 r/min、流动相流速2.0 mL/min、分离温度25℃、检测波长254 nm条件下进行分离,所得产物纯度经HPLC检测,结构经MS、1 H-NMR和13 C-NMR鉴定。4 h内从200 mg经XAD-16大孔树脂预处理的粗提物中一步分离制备得到33.5mg葫芦素B、21.7 mg葫芦素E和14.9 mg葫芦素Ⅰ,纯度分别为99.1%、98.3%和97.5%,结构经鉴定与文献相符。

A preparative high-speed counter-current chromatography（HSCCC） was used to isolate and separate cucurbitacins from Pedicellus Melo.A successful isolation and purification was achieved at the following conditions： the two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water at a volume ratio of 1.2∶0.8∶0.8∶1.2 was selected and the lower phase of the system was used as the mobile phase at the flow rate of 2.0 mL/min,the isolation temperature and revolution speed were set at 25 ℃ and 800 r/min,respectively.In 4.0 h,the isolation produced a total of 33.5 mg cucurbitacin B,21.7 mg cucurbitacin E,and 14.9 mg cucurbitacin I with purities of 99.1%,98.3% and 97.5%,respectively,determined by high performance liquid chromatography from 200 mg crude extract after cleaning-up by XAD-16 macroporous resin,which was necessary for the excellent purification.The chemical structure identification was carried out by 1H-NMR and 13C-NMR.