茶類是全球最為流行的飲品之一，因此，對於了解其中成份也是相當重要的。本研究提出以毛細管電泳結合電激發化學放光偵測法(CE-ECL)，並以緩衝溶液為分離基礎，以進行偵測其胺類化合物。電激發化學放光偵測法使用tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)32+)為放光試劑，並於自製的化學反應槽中進行化學偵測。本研究在具有電滲流(electroosmotic flow, EOF)之電泳環境下，使用緩衝溶液成功地分離出較多茶葉的胺類化合物訊號，於優化的分離條件下，最佳分離效果之緩衝溶液為100 mM Boric acid (pH 9.0)。然而，在人體體液中脯胺酸(proline)的含量，對於腎功能不全或慢性尿毒症患者是一個重要的生物參數。因此，本研究以毛細管電泳結合電激發化學放光偵測法(CE-ECL)，並以緩衝溶液為分離基礎，偵測人體血漿中的脯胺酸，並進行定量。於優化的偵測條件下，使用電壓進樣並利用樣品堆積(sample stacking)的方式，使內標準品TPA的最低濃度可偵測至1 μM，於真實樣品血漿中加入TPA，可偵測到的TPA濃度為10 μM。而標準品脯胺酸的最低濃度可偵測至1 μM，其偵測極限為0.17 μM。因此，本實驗相較於雷射誘發螢光、發光二極體誘發螢光，提供了實驗裝置簡單、實驗成本低、樣品消耗量少及高解析度等優點的分離方法。 Tea is one of the most popular beverages in the world, therefore, to understand which ingredients are also very important. This study proposed an optimized buffer solution for the separation of amine compounds by capillary electrophoresis with electrogenerated chemiluminescence(ECL)detection. The tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)32+) was ultilized for ECL on the surface of indium tin oxide (ITO) electrode that embeded in a home-made PDMS detection cell. The electrophoretic separation of amines was performed by 100 mM boric acid (pH 9.0) in the presence of electroosmotic flow (EOF).The detection of ECL was improved by electrokinetic sample stacking under optimal condition. The limit of detection of standard tripropylamine (TPA) could be reached to 1 μM while the plasma sample that spiked with TPA was riased to 10 μM, and the minimum detectable concentration of proline to 1 μM, which detection limit of 0.17 μmol/l. In this study, we demonstrated the CE-ECL is useful for the determination of amine compounds from tea and human plasma. Moreover, our data also shown the CE-ECL have enough sensitity for the determination of secondary amine from body fluid. Therefore, the CE-ECL may potentially be a powerful tool for the determination of proline in the clinical diagnosis of cancer ?
