Kostal Vratislav, Zeisbergerova Marta, Hrotekova Zuzana, Slais Karel, Kahle Vladislav
Department of Analytical Chemistry, Faculty of Science, Palacky University, Olomouc, Czech Republic.
Electrophoresis. 2006 Dec;27(23):4658-65. doi: 10.1002/elps.200600222.
A miniaturized post-column fluorimetric detection cell for capillary separation methods based on optical fibers and liquid core waveguides (LCWs) is described. The main part of the detection cell is a fused-silica capillary coated with Teflon AF serving as an LCW. The optical fibers are used both for coupling the excitation source with the detection domain in the LCW and for the axial fluorescence collection from the LCW end. The latter fiber is connected with a compact CCD spectrometer that serves for the rejection of the scattered excitation light and for the fluorescence signal detection. The proposed design offers a compact fluorescence detector for various microcolumn separation techniques without optical elements such as filters or objectives. Moreover, its construction and optical adjustment are very simple and the whole system is highly miniaturized. The function of the detection cell is demonstrated by CE of amino acids labelled by fluorescein-based tags. Separations of different standard amino acid mixtures and plasma samples are presented. The comparison of plasma amino acid levels of individuals being in good health with those of patients with inherited metabolic disorders is also shown.
描述了一种基于光纤和液芯波导(LCW)的用于毛细管分离方法的小型化柱后荧光检测池。检测池的主要部分是一根涂有特氟龙AF的熔融石英毛细管,用作液芯波导。光纤既用于将激发源与液芯波导中的检测区域耦合,也用于从液芯波导末端轴向收集荧光。后一根光纤与一台紧凑型电荷耦合器件(CCD)光谱仪相连,该光谱仪用于滤除散射的激发光并检测荧光信号。所提出的设计为各种微柱分离技术提供了一种无需滤光片或物镜等光学元件的紧凑型荧光检测器。此外,其结构和光学调整非常简单,整个系统高度小型化。通过对基于荧光素标记的氨基酸进行毛细管电泳(CE)来证明检测池的功能。展示了不同标准氨基酸混合物和血浆样品的分离情况。还展示了健康个体与遗传性代谢紊乱患者血浆氨基酸水平的比较。