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

建立了一种结合胶束电渗色谱(MEC)和激光激发荧光检测的高灵敏方法。该方法能够检测几纳升体积内金属络合物的浓度。本研究分为两个部分：(1)提供了用于MEC分离所需的螯合剂的必要条件；(2)对某些卟啉络合物的Ar~+激光激发荧光检测体系进行了初步评价。(1)对某些二羟基偶氮染料和肼类化合物的MEC保留和拆分研究表明，相对较弱的螯合物溶质与胶束拟相的相互作用对MEC的精细拆分至关重要。与胶束的紧密结合虽然加强了溶质的保留，但对MEC不利。因此，应首选电荷类型与所用胶束相同的络合物，和/或具有亲水功能的络合物，以降低向胶束中的分配常数。(2)制作了Ar-离子激光单色仪组件，以评估某些荧光剂的性能，如卟啉螯合物。使用0.5ml流动池，四(i-甲基吡啶1-4-y1)卟啉镁(II)-，，，-四(i-甲基吡啶1-4-y1)螯合物的最佳检测下限(S/H=2)被测定为3.4×10^-12；M。这个浓度换算成‘激光照射’体积中的螯合物的绝对量3.6×10^；-19；摩尔，约为107毫微升。这一数据清楚地表明，MEC与激光激发荧光检测的耦合方案确保了在6NL注射等超微量样品量下阿托莫尔的检测。因此，该系统在研究金属离子和金属络合物的迁移和去向方面具有很好的应用前景。

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.