高速逆流色谱结合制备液相色谱法分离制备葡萄籽中的多酚

采用高速逆流色谱结合制备液相色谱法从葡萄籽乙醇提取物中分离得到了8种多酚。高速逆流色谱以上相为固定相，下相为流动相，主机转速为900 r/min，流速为2 mL/min，分离温度为25℃，检测波长为280 nm，利用正向和反向洗脱相结合的模式，在正丁醇-乙酸乙酯-水（1∶14∶15，v/v/v）和正己烷-乙酸乙酯-水（1∶10∶10，v/v/v）溶剂系统下从葡萄籽提取物中分离得到了5种多酚。原花青素B₁、原花青素B₂、没食子酸、表儿茶素没食子酸酯和儿茶素的纯度分别为98.5%、97.2%、98.3%、98.9%和96.7%。利用制备液相色谱法对高速逆流色谱分离成分进一步分离纯化，获得了表儿茶素、表没食子儿茶素没食子酸酯和没食子儿茶素没食子酸酯，纯度分别99.2%、99.3%和99.2%。该方法单次制备量均达到毫克级，简便、快速、分离纯度高，适合于葡萄籽中多酚的分离制备。

Isolation and preparation of polyphenols from grape seed by high-speed counter-current chromatography combined with preparative high performance liquid chromatography

Eight polyphenols were separated from grape seed by high-speed counter-current chromatography (HSCCC). The separation was performed using a two-phase solvent system composed of 1-butanol-ethyl-water (1:14:15, v/v/v) and n-hexane-ethyl-water (1:10:10, v/v/v). The upper and lower phases were used as the stationary and mobile phases, respectively. A flow rate of 2.0 mL/min was employed for the separation. The apparatus was rotated at 900 r/min, and the detection wavelength was set at 280 nm. Under the selected conditions, procyanidins B₁, procyanidins B₂, gallic acid, epicatechin gallate, and catechin were obtained from the grape seed extracts with a purity of 98.5%, 97.2%, 98.3%, 98.9%, and 96.7%, respectively. Epicatechin, epicatechin gallate, and gallocatechin gallate were obtained by preparative high performance liquid chromatography with the purity of 99.2%, 99.3%, and 99.2%, respectively. The proposed method is simple and shows high separation efficiency, and will be of great importance for the comprehensive utilization of grape seed.