Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki, Greece.
Anal Chim Acta. 2013 Sep 17;795:75-81. doi: 10.1016/j.aca.2013.07.015. Epub 2013 Jul 12.
In this study the development, validation and application of a new chromatographic method for the determination of glutathione (GSH) in wine samples is presented. The separation of the GSH was carried out using a sulfobetaine-based hydrophilic interaction chromatography (HILIC) analytical column whereas its detection was carried out spectrofluorimetrically (λext/λem=340/455 nm) after post-column derivatization with o-phthalaldehyde. GSH was separated efficiently from matrix endogenous compounds of wines by using a mobile phase of 15 mmol L(-1) CH3COONH4 (pH=2.5)/CH3CN, 35/65% (v/v). The parameters of the post-column reaction (pH, amount concentration of the reagent and buffer solution, flow rate, length of the reaction coil) were investigated. The linear determination range for GSH was 0.25-5.0 μmol L(-1) and the LOD was 19 nmol L(-1). No matrix effect was observed, while the accuracy was evaluated with recovery experiments and was ranged between 89% and 108%.
本研究提出了一种新的色谱方法，用于测定葡萄酒样品中的谷胱甘肽（GSH）。采用基于磺丁基甜菜碱的亲水相互作用色谱（HILIC）分析柱分离 GSH，然后在柱后衍生化后用邻苯二醛进行荧光检测（λext/λem=340/455nm）。通过使用 15 mmol L(-1) CH3COONH4（pH=2.5）/CH3CN，35/65%（v/v）作为流动相，GSH 可以有效地从葡萄酒的基质内源性化合物中分离出来。考察了柱后反应的参数（pH、试剂和缓冲溶液的浓度、流速、反应盘管的长度）。GSH 的线性测定范围为 0.25-5.0 μmol L(-1)，LOD 为 19 nmol L(-1)。未观察到基质效应，而准确性则通过回收实验进行评估，回收率在 89%至 108%之间。
