STUDY OF ZONE BROADENING IN OPTICALLY GATED HIGH-SPEED CAPILLARY ELECTROPHORESIS
STUDY OF ZONE BROADENING IN OPTICALLY GATED HIGH-SPEED CAPILLARY ELECTROPHORESIS
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DOI：
10.1021/ac00072a004
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发表时间：
1993-12-15
影响因子：
7.4
通讯作者：
JORGENSON, JW
中科院分区：
文献类型：
--
作者：
MOORE, AW;JORGENSON, JW
异硫氰酸荧光素（FITC）标记的化合物通过毛细管区带电泳（CZE）在几秒而不是几分钟内分离，同时保持高分离效率。这些快速的分析时间是通过在短的分离距离上施加高电场和使用独特的光学门控进样系统来实现的。光选通注入是基于由氩离子激光束诱导的FITC的光分解。该相同激光束的一部分也用于通过柱上激光诱导荧光检测分析物区域。四个FITC标记的氨基酸和荧光素染料进行了分析，比较系统性能的理论预测。为此，通过实验测量每个分析物区的总方差。将其与进样和检测系统以及纵向扩散的理论预期方差贡献之和进行比较。由于纵向扩散的方差计算扩散常数实验测量与传统的CZE系统。在考虑了这三个已知的方差来源后，发现该方法在2-3 s的分析时间内实现了理论预测性能的80%以上。
Fluorescein isothiocyanate (FITC) labeled compounds are separated by capillary zone electrophoresis (CZE) in seconds rather than minutes while high separation efficiency is maintained. These rapid analysis times are achieved through high electric fields applied over short separation distances and the use of a unique optical-gating injection system. The optical-gating injection is based on photodecomposition of FITC induced by an argon ion laser beam. A fraction of this same laser beam is also used for detection of the analyte zones by on-column laser-induced fluorescence. Four FITC-labeled amino acids and fluorescein dye were analyzed to compare system performance to that predicted by theory. To do so, the total variance for each analyte zone was measured experimentally. This was compared to the sum of the theoretically expected variance contributions from the injection and detection systems and longitudinal diffusion. The variance due to longitudinal diffusion was calculated from diffusion constants measured experimentally with a traditional CZE system. After accounting for these three known sources of variance, this method is found to achieve better than 80% of the performance predicted by theory in analysis times as short as 2-3 s.