Zhao Shulin, Huang Yong, Shi Ming, Liu Yi-Ming
College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 51004, China.
J Chromatogr A. 2009 Jun 26;1216(26):5155-9. doi: 10.1016/j.chroma.2009.04.081. Epub 2009 May 3.
A highly sensitive microchip electrophoresis (MCE) method with chemiluminescence (CL) detection was developed for the determination of biogenic amines including agmatine (Agm), epinephrine (E), dopamine (DA), tyramine, and histamine in human urine samples. To achieve a high assay sensitivity, the targeted analytes were pre-column labeled by a CL tagging reagent, N-(4-aminobutyl)-N-ethylisoluminol (ABEI). ABEI-tagged biogenic amines after MCE separation reacted with hydrogen peroxide in the presence of horseradish peroxidase (HRP), producing CL emission. Since no CL reagent was added to the running buffer, the background of the CL detection was extremely low, resulting in a significant improvement in detection sensitivity. Detection limits (S/N=3) were in the range from 5.9x10(-8) to 7.7x10(-8) M for the biogenic amines tested, which were at least 10 times lower than those of the MCE-CL methods previously reported. Separation of a urine sample on a 7 cm glass/poly(dimethylsiloxane) (PDMS) microchip channel was completed within 3 min. Analysis of human urine samples found that the levels of Agm, E and DA were in the ranges of 2.61x10(-7) to 4.30x10(-7) M, 0.81x10(-7) to 1.12x10(-7) M, and 8.76x10(-7) to 11.21x10(-7) M (n=4), respectively.
开发了一种具有化学发光（CL）检测功能的高灵敏度微芯片电泳（MCE）方法，用于测定人尿液样本中的生物胺，包括胍丁胺（Agm）、肾上腺素（E）、多巴胺（DA）、酪胺和组胺。为了实现高检测灵敏度，目标分析物通过CL标记试剂N-(4-氨丁基)-N-乙基异鲁米诺（ABEI）进行柱前标记。MCE分离后的ABEI标记生物胺在辣根过氧化物酶（HRP）存在下与过氧化氢反应，产生CL发射。由于运行缓冲液中未添加CL试剂，CL检测的背景极低，从而显著提高了检测灵敏度。所测试生物胺的检测限（S/N=3）在5.9×10⁻⁸至7.7×10⁻⁸M范围内，至少比先前报道的MCE-CL方法低10倍。在7 cm玻璃/聚二甲基硅氧烷（PDMS）微芯片通道上对尿液样本的分离在3分钟内完成。对人尿液样本的分析发现，Agm、E和DA的水平分别在2.61×10⁻⁷至4.30×10⁻⁷M、0.81×10⁻⁷至1.12×10⁻⁷M和8.76×10⁻⁷至11.21×10⁻⁷M范围内（n=4）。
