The Development of Novel Immunoassay System using fluorescent Rareearth Metal Complex Labels and Transparent Polymer Films.

这项工作是通过结合使用长寿命稀土配合物标签和透明聚合物薄膜来开发时间分辨荧光免疫测定系统。这项工作的中心目标是；(1)建立具有长寿命“能量转移荧光”的螯合配体的分子设计标准；(2)研究可塑聚氯乙烯薄膜作为分离和荧光增强的介质。主要研究结果如下：(1)利用脉冲氮激光辐射源（337.1 nm）完成了时间分辨荧光测量仪的研制。用乙醇中苯甲酰三氟丙酮铕配合物的模型发光体系验证了该系统的准确性和时间分辨能力。(2)八齿酸多胺羧酸配体已被商业化用于生物系统中钙离子的检测，作为镧系离子的能量转移荧光标记的有希望的候选者。它们具有喹啉环或苯基环的“天线基团”以吸收近紫外光，并具有N， o给体集，足以满足镧系离子的配位数。(3)用氯仿从pH为10的碱性溶液中提取出强荧光螯合离子对Q[Eu(tta)_4] （tta：三氟乙酸乙酯，Q^+：季铵离子），其荧光强度比母体配合物[Eu(tta)_3]的荧光强度大100倍。(4)用二辛基膦酸酯增塑PVC薄膜对Q[Eu(tta)_4]的荧光增强作用不大，相反，它增强了三苯基甲烷染料乙基紫的荧光强度。这一观察结果表明，荧光孔与薄膜介质之间的匹配是设计这类化学系统时考虑的一个关键因素。

This work has been conducted for the exploitation of time-resolved fluoimetric immunoassay systems by the combined use of long-lived rare-earth complex labels and transparent polymer films. The central aims of this work are ; (1) to establish the molecular designing criteria for the chelating ligands which give the long-lived 'energy transfer fluorescence' and (2) to examine plasticized polyvinyl chloride films as the media for the separation and the fluorescence enhancement. Main results are outlined as :(1) Time-resolved fluorescence measurement apparatus was accomplished wiht a pulsed nitrogen laser radiation source (337.1 nm). The accuracy and time-resolution capability of the builtup system was confirmed using the model luminescence system of benzoyltrifluoroacetonato Eu complex in ethanol.(2) The octadentate polyminocaboxylate ligands which have been Commercially available for calcium ion detection in biological systems have been proposed as promising candidates for the energy transfer fluorescence labels for the lanthanide ions. They have the 'antenna groups' of quinolyl or phenyl rings for absorption of near uv light and posses the N, O-donor set sa suffices for the coordination number of the lanthanide ions.(3) The strongly fluorescent chelate ion-pair Q[Eu(tta)_4] has been found to extracted in chloroform from alkaline solution at pH 10 (tta : thenoyltrifluoroacetonate, Q^+ : quaternary ammonium ion), whose fluorescence intensity was by a factor of 100 greater than that of the parent complex, [Eu(tta)_3].(4) A PVC film plasticized with dioctylphhalate was unfortunately not so beneficial to the fluorescence enhancement of Q[Eu(tta)_4], by contrast, it enhances the intensity of a triphenylmethane dye, ethyl violet. This observation indicates that the matching between fluorescent folutes and the film medium is a crucial factor to be conceived on the designing of such chemical systems for the luminescence in somewhat ambiguous nature.