高温二维液相色谱系统的构建及其评价

在二维液相色谱中, 第二维的分离速度是制约其发展的重要因素. 升高色谱柱温度可以有效降低流动相粘度, 加快溶质在两相间的传质速率, 有效加快分析速度. 以离子交换色谱法(WAX)为第一维分离模式和反相色谱法(RP)为第二维分离模式, 十通阀和两个捕集柱为接口, 通过将第二维色谱柱温度升高到80 ℃和提高流量到3 mL/min, 构建了高温WAX/RP二维液相色谱系统. 以4种标准蛋白的酶解物为样品评价系统的分离性能, 第一维共有33个馏分被捕集并导入到第二维分析, 高温二维液相色谱系统识别出187个色谱峰.

Construction and Evaluation of High Temperature Two-Dimensional Liquid Chromatography System

The separation speed is very important .to the development of two dimensional liquid chroma-tography. The viscosity of the mobile phase decreases and the analyte diffusivity increases at higher tem-perature. An ion exchange chromatography column was used as the first dimension mode and reversed phase chromatography as the second dimension mode, which were connected by the interface consisting of a ten-port valve and two trap columns. A high temperature two dimensional weak anion exchange (WAX)/ reversed phase (RP) liquid chromatography system was developed by the second dimensional column oper-ated at 80℃ and higher flow rate up to 3 mL/min. A tryptic digest of 4 proteins was separated on this sys-tem. 33 fractions from the first dimension were introduced to the second dimension by the interface. 187 peaks were separated by the high temperature two dimensional liquid chromatography system.