DEVELOPMENT OF CHEMICAL SYSTEMS FOR SEPARATION AND PURIFICATION BY MICROCHIP ELECTROPHORESIS

本项目的目的是开发一种将微芯片电泳(MCE)作为一种高效分离纯化技术的化学系统。取得的主要结果如下：1。将SRW、SRMM、SCOVING等在线样品预富集技术应用于微芯片胶束电动色谱(MCMEKC)，设计了一种新的通道构型，制作了一些微芯片。通常，通过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。研究了在MCMEKC中使用热透镜显微镜(TLM)作为一种高灵敏度的检测方案，并结合扫描技术，观察到灵敏度提高了近500倍。利用液结系统和喷雾芯片或内径较窄的短毛细管，研制了一种用于MCE-MS系统的电喷雾电离(ESI)接口，实现了稳定的电离和减小的谱带展宽。在MCE-MS系统中，成功地分离和检测到了多个多肽，为生物源化合物的分析奠定了基础。在微芯片电色谱分离中，不饱和聚酯及其共聚物和聚二甲基硅氧烷(PDMS)等材料被用来制备微芯片，在制备通道时加入了一些功能和反应器。采用集成在PDMS芯片中的LED作为光源，获得了良好的检测性能。通过光化学反应固定化酶，成功地制备了微通道中特定位置修饰的微反应器芯片，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.