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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.

本文建立了胶束电渗色谱-激光荧光检测法。该方法能够在几纳升体积中检测阿托摩尔水平的金属螯合物。本研究工作分两部分进行：（1）提供了在MEC分离中所需的螯合试剂的条件;（2）对某些卟啉螯合物的氩离子激光激发荧光检测体系进行了初步评价。(1)一些二羟基偶氮染料和腙的MEC保留和分辨率的研究清楚地表明，相对较弱的相互作用的chalate溶质与胶束假相是至关重要的微妙的MEC分辨率。与胶束的紧密结合虽然会降低溶质的保留，但对MEC不利。因此，应优选与所用胶束具有相同电荷类型的螯合物，和/或具有降低胶束中分配常数的亲水性官能团的螯合物。(2)氩离子激光单色仪装置已被用来评估某些荧光剂如卟啉螯合物的可探测性。使用0.5 ml流动池，测得Mg（II）-，，-四（1-甲基吡啶-4-基）卟吩螯合物的检测限（S/H = 2）为3.4 x 10 μ <-12>M。该浓度被转化为螯合物的绝对量，<-19>在“激光照射”体积中为3.6 x 10 μ mol，约为1.5 μ mol。107 nL。该数据清楚地表明，与激光激发荧光检测耦合的MEC方案确保了在超微量样品量（例如6 nL进样）中的阿托摩尔检测。因此，该系统是有前途的生物研究的转运和命运的金属离子和/或金属配合物。