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Development of High Performance Micellar Electroosmotic Chromatography-Laser Excited Fluorescence Detection Scheme.

高性能胶束电渗色谱-激光激发荧光检测方案的开发。

基本信息

批准号：
62850137
负责人：
YOTSUYANAGI Takao
金额：
$ 5.12万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

项目摘要

A highly sensitive method has been established with a combined use of miceller electroosmotic chromatographyt (MEC) with a laser-excited fluorescence detection. The method is capable of detecting attomol level of metal chelates in a few nanoliter volume. This reseach project has been conducted to two parts; (1) the provision of requisite conditions for the chelating reagents which ought to be satisfied in the MEC separation, (2) a preliminary evaluation of the argon-ion laser excited fluorescence detction system for some porphyrin chelates.(1) The MEC retention and resolution studies for some dihydroxyazo dyes and hydrazones clearly indicated that the relatively weak interaction of chalate solute with micellar pseudo phase is crucial for the delicate MEC resolution. The tight binding to the micelles, thouht strengtens the solute retention, is unfavorable for the MEC. Hence, the chelates of the same charge type as that of the micelles emplyed, and/or the chelates bearing hydrophilic functions which lower the patition constants into the micelles should be preferred.(2) The argon-ion laser - monochrometer assembly has been made to assess the betectability for some fluorescers such as porphyrin chelates. Using a 0.5 ml flow cell, the betection limit (S/H = 2) of Mg(II)- , , , -tetrakis(I-methylpyridy1-4-y1) porphine chelate is determined to be 3.4 x 10^<-12> M. This concentration is translated into the absolute amount of the chelate, 3.6 x 10^<-19> mol in a 'laser-irradiated' volume, ca. 107 nL. This data clearly show that the coupled MEC scheme with the laser-excited fluorescence detection ensures the attomol detection in the ultramicro sample quantities such as a 6 nL injection. Consequently, the system is promising for biological study of tranport and fate of metal ions and/or metal complexes.

建立了胶束电渗透色谱（MEC）与激光激发荧光相结合的高灵敏度检测方法。该方法能够检测几纳升体积内金属螯合物的原子浓度。本研究项目共分为两部分；(1)提供了MEC分离中应满足的螯合试剂的必要条件；(2)氩离子激光激发荧光检测系统对部分卟啉螯合物的初步评价。(1)对一些二羟基偶氮染料和腙的MEC保留和分解研究清楚地表明，chalate溶质与胶束伪相相对弱的相互作用是精细MEC分解的关键。与胶束的紧密结合虽然加强了溶质的保留，但对MEC是不利的。因此，与所使用的胶束具有相同电荷类型的螯合物和/或具有降低胶束分配常数的亲水性功能的螯合物应该是首选的。(2)利用氩离子激光单色仪组件对某些荧光剂（如卟啉螯合物）的可选择性进行了评价。使用0.5 ml流动池，确定Mg(II)-,,， -四（i -甲基吡啶-1 -4-y1）卟啉螯合物的检测极限（S/H = 2）为3.4 × 10^<-12> mol .该浓度转化为螯合物的绝对量，3.6 × 10^<-19> mol在“激光照射”的体积中，约为107 nL。这一数据清楚地表明，耦合MEC方案与激光激发荧光检测保证了在6 nL注射等超微样品量下的原子醇检测。因此，该系统有望用于金属离子和/或金属配合物的运输和命运的生物学研究。