胶束电动色谱联用新型多光子激发荧光检测法分离与测定氨基酸

Separation and determination of amino acids by micellar electrokinetic chromatography coupling with novel multiphoton excited fluorescence detection.

作者信息

Chen Sheng, Xu Youzhi, Xu Fei, Feng Xiaojun, Du Wei, Luo Qingming, Liu Bi-Feng

机构信息

The Key Laboratory of Biomedical Photonics of MOE - Hubei Bioinformatics and Molecular Imaging Key Laboratory, Department of Systems Biology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.

出版信息

J Chromatogr A. 2007 Aug 31;1162(2):149-53. doi: 10.1016/j.chroma.2007.05.046. Epub 2007 May 18.

DOI:10.1016/j.chroma.2007.05.046

PMID:17560588

Abstract

In this article, it was demonstrated that separation and determination of 20 amino acids were accomplished by micellar electrokinetic chromatography (MEKC) coupling with novel multiphoton excited fluorescence (MPEF) detection method. Different from MPEF achieved by expensive fs laser, continuous wave (CW) diode laser of ultra-low cost was uniquely employed in our MPEF system. Amino acids were fluorescently labeled with fluorescein isothiocyanate (FITC), and were subjected to sodium dodecyl sulfate (SDS)-based MEKC separation and CW-based MPEF detection. The result was compared with that by single photon excited fluorescence (SPEF), which indicated that MPEF had the advantages of better mass detectability and higher separation selectivity over SPEF. Quantitative analysis was performed and revealed linear dynamic range of over 2 orders of magnitude, with mass detection limit down to ymole level. To evaluate the reliability, this method was successfully applied for analyzing a commercial nutrition supplement liquid.

摘要

在本文中，通过胶束电动色谱（MEKC）与新型多光子激发荧光（MPEF）检测方法联用实现了20种氨基酸的分离与测定。与使用昂贵的飞秒激光实现的多光子激发荧光不同，我们的多光子激发荧光系统独特地采用了超低成本的连续波（CW）二极管激光器。氨基酸用异硫氰酸荧光素（FITC）进行荧光标记，然后进行基于十二烷基硫酸钠（SDS）的胶束电动色谱分离和基于连续波的多光子激发荧光检测。将结果与单光子激发荧光（SPEF）的结果进行比较，结果表明多光子激发荧光在质量检测能力和分离选择性方面优于单光子激发荧光。进行了定量分析，结果显示线性动态范围超过2个数量级，质量检测限低至飞摩尔水平。为评估该方法的可靠性，将其成功应用于分析一种商业营养补充液。