基于 HPLC 荧光法测定葡萄酒中谷胱甘肽衍生物的含量。

A HPLC fluorescence-based method for glutathione derivatives quantification in must and wine.

机构信息

UMR 1219 Oenologie, ISVV, Université de Bordeaux, 210, Chemin de Leysotte, CS 50008, 33 882 Villenave d'Ornon Cedex, France.

出版信息

Anal Chim Acta. 2010 Feb 15;660(1-2):158-63. doi: 10.1016/j.aca.2009.09.042. Epub 2009 Oct 2.

DOI:10.1016/j.aca.2009.09.042

PMID:20103157

Abstract

A simple and automated high performance liquid chromatography (HPLC) method for the separation and quantitative determination of gamma-glutamylcysteine and reduced glutathione (GSH) in wines is reported. This technique involves the use of a pre-column derivatization with 2,3-naphthalenedialdehyde (NDA), an isocratic separation in presence of beta-cyclodextrine and a fluorimetric detection. The quantification of oxidized glutathione (GSSG) has also been studied, for the first time in wines, using an additive pre-derivatization step for reduction using glutathione reductase. The method has been designed for use in laboratories with limited equipment. The assay has been optimized and presents very good performances in terms of sensitivity and selectivity. Then, it has been validated for linearity, LOD, LOQ, precision and accuracy.

摘要

本文报道了一种简单、自动化的高效液相色谱（HPLC）方法，用于分离和定量测定葡萄酒中的γ-谷氨酰半胱氨酸和还原型谷胱甘肽（GSH）。该技术涉及使用 2,3-萘二醛（NDA）进行柱前衍生化，在β-环糊精存在下进行等度分离，并进行荧光检测。还首次研究了葡萄酒中氧化型谷胱甘肽（GSSG）的定量，使用谷胱甘肽还原酶进行还原的添加剂预衍生化步骤。该方法专为设备有限的实验室设计。该测定法已进行了优化，在灵敏度和选择性方面具有非常好的性能。然后，对其线性、LOD、LOQ、精密度和准确度进行了验证。