Tsai H C, Whang C W
Department of Chemistry, Tunghai University, Taichung, Taiwan.
Electrophoresis. 1999 Sep;20(12):2533-8. doi: 10.1002/(SICI)1522-2683(19990801)20:12<2533::AID-ELPS2533>3.0.CO;2-#.
A capillary electrophoresis (CE)/indirect chemiluminescence (CL) detection method is described for monoamines, viz., serotonin (5-HT), dopamine (DA), epinephrine (EP), and norepinephrine (NE) and for catechol (CA). Optimal separation and detection were obtained with an electrophoretic buffer of 10 mM sodium borate (pH 9.5) containing 5 mM luminol and 25 mM H2O2, and a catalyst solution of 30 microM CuSO4 in 30 mM borate buffer (pH 10.0). Complete separation of 5-HT, DA, EP, NE and CA was achieved in less than 5 min. The Cu(II)-catalyzed luminol CL reaction was employed to provide the high and constant background. Since monoamines and catechol can form stable complexes with Cu(II), inverted analyte peaks due to decreased catalytic activity of Cu(II) can be detected. The degree of CL suppression is proportional to the analyte concentrations. Linearity (r> or =20.99) over two orders of magnitude was generally obtained. The concentration limits of detection (CLODs) for the monoamines and catechol studied were between 0.5 and 3.1 uM. The relative standard deviation (RSD) values on peak size and migration time were in the ranges 3.2-4.4% and 0.4-0.5%, respectively. The applicability of the method for the analysis of pharmaceutical and biological samples was examined.
本文描述了一种用于单胺类物质（即血清素（5-羟色胺，5-HT）、多巴胺（DA）、肾上腺素（EP）和去甲肾上腺素（NE））以及儿茶酚（CA）的毛细管电泳（CE）/间接化学发光（CL）检测方法。使用含有5 mM鲁米诺和25 mM过氧化氢的10 mM硼酸钠（pH 9.5）电泳缓冲液以及30 mM硼酸盐缓冲液（pH 10.0）中30 microM硫酸铜的催化剂溶液，可实现最佳分离和检测。5-HT、DA、EP、NE和CA在不到5分钟内实现了完全分离。采用铜（II）催化的鲁米诺CL反应提供高且恒定的背景。由于单胺类物质和儿茶酚可与铜（II）形成稳定络合物，因此可检测到因铜（II）催化活性降低而出现的反向分析物峰。CL抑制程度与分析物浓度成正比。通常在两个数量级上获得线性关系（r≥0.99）。所研究的单胺类物质和儿茶酚的检测浓度限（CLODs）在0.5至3.1 μM之间。峰面积和迁移时间的相对标准偏差（RSD）值分别在3.2 - 4.4%和0.4 - 0.5%范围内。考察了该方法在药物和生物样品分析中的适用性。
