Quantification of biogenic amines by microchip electrophoresis with chemiluminescence detection.
Quantification of biogenic amines by microchip electrophoresis with chemiluminescence detection.
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DOI：
10.1016/j.chroma.2009.04.081
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发表时间：
2009-06-26
期刊：
影响因子：
--
通讯作者：
Liu YM
中科院分区：
文献类型：
--
作者：
Zhao S;Huang Y;Shi M;Liu YM
建立了一种高灵敏度的微芯片电泳-化学发光检测法（MCE-CL），用于人尿样中胍丁胺、肾上腺素、多巴胺、酪胺和组胺等生物胺的测定。为了实现高测定灵敏度，目标分析物通过CL标记试剂N-（4-氨基丁基）-N-乙基异鲁米诺（ABEI）进行柱前标记。ABEI标记的生物胺MCE分离后与过氧化氢在辣根过氧化物酶（HRP）的存在下反应，产生CL发射。由于运行缓冲液中未加入CL试剂，CL检测的背景极低，导致检测灵敏度显著提高。检测限（S/N =3）范围为5.9 × 10−8至7.7 × 10−8 M的生物胺测试，这是至少10倍低于先前报道的MCE-CL方法。尿液样本在7 cm玻璃/聚二甲基硅氧烷（PDMS）微芯片通道上的分离在3分钟内完成。分析人类尿液样本发现，Agm、E和DA的水平范围分别为2.61 ×10−7至4.30×10−7 M、0.81×10−7至1.12×10−7 M和8.76×−7至11.21×10−7 M（n=4）。
A highly sensitive microchip electrophoresis (MCE) method with chemiluminescence (CL) detection was developed for the determination of biogenic amines including agmatine, epinephrine, dopamine, 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 5.9 × 10−8 to 7.7 × 10−8 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 2.61 ×10−7 to 4.30×10−7 M, 0.81×10−7 to 1.12×10−7 M, and 8.76×−7 to 11.21×10−7 M (n=4), respectively.
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