Beard Nigel P, de Mello Andrew J
AstraZeneca/SmithKline Beecham Centre for Analytical Sciences, Imperial College of Science, Technology and Medicine, Department of Chemistry, South Kensington, London, UK.
Electrophoresis. 2002 Jun;23(11):1722-30. doi: 10.1002/1522-2683(200206)23:11<1722::AID-ELPS1722>3.0.CO;2-W.
A polydimethylsiloxane-glass capillary microchip is fabricated for the rapid analysis of a mixture of common biogenic amines using indirect fluorescence detection. Using a running buffer of phosphate and 2-propanol, and Rhodamine 110 as a background fluorophore, both co-ionic and counter-ionic systems are explored. Studies demonstrate the separation and analysis of cations using indirect fluorescence detection for the first time in a chip-based system. Resulting electrophoretic separations are achieved within a few tens of seconds with detection limits of approximately 6 microM. The reduced sample handling and rapid separations afforded by the coupling of indirect fluorescence detection with chip-based capillary electrophoresis provide a highly efficient method for the analysis and detection of molecules not possessing a chromophore or fluorophore. Furthermore, limits of detection are on a par with reported chip-based protocols that incorporate precolumn derivatisation with fluorescence detection. The current device circumvents lengthy sample preparation stages and therefore provides an attractive alternative technique for the analysis biogenic amines.
制备了一种聚二甲基硅氧烷-玻璃毛细管微芯片,用于使用间接荧光检测法快速分析常见生物胺混合物。使用磷酸盐和2-丙醇的运行缓冲液以及罗丹明110作为背景荧光团,研究了共离子和反离子系统。研究首次在基于芯片的系统中使用间接荧光检测法对阳离子进行了分离和分析。几十秒内即可实现电泳分离,检测限约为6 microM。间接荧光检测与基于芯片的毛细管电泳相结合,减少了样品处理量并实现了快速分离,为分析和检测不具有发色团或荧光团的分子提供了一种高效方法。此外,检测限与报道的采用柱前衍生化荧光检测的基于芯片的方法相当。当前的装置避免了冗长的样品制备阶段,因此为生物胺分析提供了一种有吸引力的替代技术。