DETECTION OF BIOMOLECULES BY CHEMILUMINESCENCE

我们已经报道了一种还原的电化学发光反应 含三(联吡啶)Ru(III)(Ru(Bpy)3+3)的试剂在 范围，而不仅仅是看到为数不多的化合物，如肼或草酸盐。 特别是，单烷基胺、二烷基胺和三烷基胺可以在 以nmole到pmole级别表示的顺序。以便对此进行便捷的研究 快速(&lt；1秒)反应，我们已经改装了一台高效液相荧光仪，用于后 柱混合。它能在0.04Pmoles检测到三丙胺。 在紫外线中难以检测到的药物，如红霉素和 在ppb水平上很容易测定克林霉素-2-磷酸。三 在这些已完成项目的基础上，提出了研究领域。第一, 为了让更快的启动和更全面的操作，我们计划在- ESA库仑流法研究Ru(Bpy)3+2氧化成Ru(BYP)3+3 手机。流动相的电化学前处理以降低检测 限制也可以用另一个库仑保护电池来实现。为了增强 化学发光寿命、阳离子、阴离子、两性离子和非离子 表面活性剂也将用于产生胶束溶液。第二， Ru(Bpy)_(3+)化学发光机理的基础研究 与胺和硫化物的反应。大口径 毛细管气相色谱将用于对可疑产品进行定量。 最后，利用高效液相色谱检测技术在分析应用中的应用 生物分子，如氨基酸、多肽和蛋白质被拼写出来。 尤其是次生氨基酸，如脯氨酸和羟脯氨酸 胶原蛋白等蛋白质将成为重点研究对象。

We have reported that the electrochemiluminescence reaction of a reducing agent with tris (bipyridine) ruthenium (III) (Ru(bpy)3+3) is more broad in scope and not just seen for a few compounds such as hydrazines or oxalate. In particular, mono-, di-, and tri-alkyl amines can be detected in the order indicated at nmole to pmole levels. To allow facile study of this fast (<1 sec) reaction, we have modified an HPLC fluorometer for post- column mixing. It is capable of detecting tripropylamine at 0.04 pmoles. Pharmaceuticals difficult to detect in the UV such as erythromycin and clindamycin-2-phosphate have been assayed easily at the ppb level. Three areas of study are proposed building on these completed projects. First, to permit faster start-up and more versatile operation, we plan to do on- line oxidation of Ru(bpy)3+2 to Ru(byp)3+3 using the ESA coulometric flow cell. Electrochemical pretreatment of the mobile phase to lower detection limits can also be done with another coulometric guard cell. To enhance chemiluminescence lifetime, cationic, anionic, zwitterionic, and nonionic surfactants will also be used to produce a micellar solution. Secondly, fundamental studies of Ru(bpy)3+3 chemiluminescence involving the mechanism of this reaction with both amines and sulfur compounds. Wide bore capillary GC will be used to quantify the suspected products. Finally, analytical applications using HPLC for the determination of biomolecules such as amino acids, peptides, and proteins are spelled out. In particular, secondary amino acids such as proline and hydroxyproline in proteins such as collagen will be focused on.