DEVELOPMENT OF CHEMICAL SYSTEMS FOR SEPARATION AND PURIFICATION BY MICROCHIP ELECTROPHORESIS

本项目的目的是开发一种化学系统，其中微芯片电泳（MCE）作为一种高性能的分离和纯化技术。主要结果如下：1。为了将SRW、SRMM和扫描等几种在线样品预富集技术应用于微芯片胶束电动色谱（MCMEKC）中，制备了一些微芯片，并设计了新的通道几何形状。通常，使用新设计的微芯片，SRW-MCMEKC.2的检测灵敏度提高了10到100倍。使用热透镜显微镜（TLM）作为MCMEKC的高灵敏度检测方案，与扫描相结合，观察到灵敏度提高了近500倍。采用液结系统和窄内径的喷雾芯片或短毛细管，建立了用于MCE-MS系统的电喷雾电离（ESI）界面，实现了稳定的电离，减小了谱带展宽。在MCE-MS系统中，成功地分离和检测了几种多肽，因此，该系统将用于分析生物源性化合物。对于微芯片电色谱（MCCEC）分离，几种材料，如不饱和聚酯及其共聚物和聚二甲基硅氧烷（PDMS），用于制备微芯片，其中一些功能和反应器在制造通道时加入。采用集成在PDMS微芯片中的LED作为光源，获得了良好的可探测性。成功制备了一种微反应器芯片，通过光化学反应固定化酶修饰微通道中的特定位点，并对其应用进行了研究。

The aim of this project was to develop a chemical system in which microchip electrophoresis (MCE) was used as a high performance separation and purification techniques. The major results obtained were as follows1. To applying several on-line sample preconcentration techniques, such as SRW, SRMM and sweeping, to microchip micellar electrokinetic chromatography (MCMEKC), some microchips, where a new channel geometry was newly designed, were fabricated. Typically, by using the new designed microchip, 10 to 100-fold increase in detection sensitivity was obtained in SRW-MCMEKC.2. The use of a thermal lens microscope (TLM) was investigated as a highly sensitive detection scheme in MCMEKC in combination with sweeping, Almost 500-fold improvement in sensitivity was observed.3. An electrospray ionization (ESI) interface for an MCE-mass spectrometry (MCE-MS) system was developed by using a liquid-junction system and a spray chip or a short capillary with narrow inner diameter, which enabled the stable ionization and reduced band broadening. In the MCE-MS system, several peptides were successfully separated and detected and hence, the system will be useful for analyzing the biogenic compounds.4. For microchip electrochromatography (MCCEC) separations, several materials, such as unsaturated polyesters and their copolymers and poly (dimethylsiloxane) (PDMS), were used for preparing microchips, where some functions and reactors were incorporated when fabricating channels. An LED incorporated in a PDMS microchip was used as a light source and good detectability was obtained. A micro-reactor chip, in which a specific site in the microchannel was modified by the immobilization of an enzyme by a photochemical reaction, was successfully prepared and its application was studied.