Sensitive detection of CZE and analysis of ultratrace biological substances by laser-induced capillary vibration method.

我们发现了激光诱导的毛细管振动（CVL）效应。CVL的产生机理是基于光热现象。试样对剪切光束的吸收引起的周期性热产生使毛细管像弦振动一样振动。CVL的振幅与样品的吸光度成正比，我们已经证明了使用CVL效应进行超微观分析。另一方面，毛细管区带电泳（CZE）被各个技术和科学领域期望成为一种强大的超微量样品从pl到nl水平的化学分离技术。然而，CZE抑制其实际应用的一个严重问题是缺乏对毛细管中分离的痕量化学物质的检测方法。我们提出将CVL应用于CZE的灵敏探测器，并取得了良好的效果。在此基础上，本课题的研究目标是开发CZE/CVL系统，并证明其优异的性能、超高的灵敏度和广泛的应用，这是初步实验所期望的。我们可以实现我们的三年研究计划。结果概要总结如下。1)设计并制作了CZE/CVL试验系统。该系统的灵敏度比传统的吸收检测器至少高2-3个数量级。2)阐明了光热现象在毛细管中产生的机理，获得了灵敏定量检测的最佳检测条件。这些条件对实际系统的设计是有用的。3)该系统可分离和定量检测氨醇水平的非衍生化氨基酸。4)分离检测吸附在聚苯乙烯微颗粒上的微量免疫球蛋白。通过优化反应条件和检测条件，有望实现亚氨基酚水平的免疫测定。

We have found a laser-induced capillary vibration (CVL) effect. The generation mechanism of CVL is based on photothermal phenomenon. Periodical heat generation caused by chopped beam absorption by the sample made the capillary vibrate like a string vibration. The amplitude of the CVL was proportional to the absorbance of the sample, and we have demonstrated ultra-micro analysis using the CVL effect. Onthe other hand, capillary zone electrophoresis (CZE) is expected from various technology and science fields to be a powerful chemical separation technique for ultra-micro quantity samples from pl to nl level. However, a severe problem with CZE inhibiting its practical uses is the lack of a detection method for trace chemical species separated in the capillary. The we have proposed to apply CVL to a sensitive detector of CZE, and shown promising results.On the basis of this course of study, the present research project aimed to develop CZE/CVL system, and to prove superior properties, ultrahigh sensitivity and wide use, which were expected from the preliminary experiments. We could attain our 3 years research plan. The outline of the results are summarized as follows.1) A trial system of CZE/CVL was designed and produced. The sensitivity of the system was at least 2-3 orders superior to conventional absorption detectors.2) The mechanisms of photothermal phenomena induced in a capillary were clarified, and an optimal detection conditions for sensitive and quantitative detection were obtained. These conditions were useful for design of a practical system.3) Non-derivatized amino acids in amol levels were separated and quantitatively detected by this system.4) Trace immunoglobulins in amol levels adsorbed on polystirene microparticles were separated and detected. By optimizing the reaction and detection conditions, sub-amol levels immunoassay are expected to be attained.