微芯片电泳-激光诱导荧光检测法分析生物样品中的 D-酪氨酸。

Trace analysis of D-tyrosine in biological samples by microchip electrophoresis with laser induced fluorescence detection.

机构信息

College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Nov 1;879(29):3203-7. doi: 10.1016/j.jchromb.2011.01.033. Epub 2011 Feb 4.

DOI:10.1016/j.jchromb.2011.01.033

PMID:21342793

Abstract

A rapid and sensitive microchip electrophoresis (MCE) method with laser induced fluorescence (LIF) detection has been developed for the quantification of D-tyrosine (Tyr) in biological samples. The assay was performed using a MCE-LIF system with glass/poly(dimethylsiloxane) (PDMS) hybrid microchip after pre-column derivatization of amino acids with fluorescein isothiocyanate (FITC). Chiral separation of the derivatives was achieved by cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) using γ-CD as chiral selector in the running buffer. D/L-Tyr enantiomer was well separated in less than 140s. The limit of detection (S/N=3) was 3.3 × 10(-8) M. Using the present method, D-Tyr level in human plasma was found to vary significantly from normal humans to patients suffering from renal failure.

摘要

建立了一种快速灵敏的微芯片电泳（MCE）-激光诱导荧光（LIF）检测方法，用于生物样品中 D-酪氨酸（Tyr）的定量分析。该方法采用 MCE-LIF 系统，在玻璃/聚二甲基硅氧烷（PDMS）混合微芯片上，在氨基酸与荧光素异硫氰酸酯（FITC）预柱衍生后进行。手性分离通过环糊精修饰的胶束电动色谱（CD-MEKC）实现，在运行缓冲液中使用γ-CD 作为手性选择剂。D/L-Tyr 对映体在不到 140s 的时间内得到很好的分离。检测限（S/N=3）为 3.3×10(-8) M。使用该方法，发现肾功能衰竭患者的人血浆中 D-Tyr 水平与正常人有显著差异。