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Published on: March 15, 2017
A HPLC fluorescence-based method for glutathione derivatives quantification in must and wine
Stéphanie Marchand1, Gilles de Revel
1UMR 1219 Oenologie, ISVV, Université de Bordeaux, 210, Chemin de Leysotte, CS 50008, 33 882 Villenave d'Ornon Cedex, France. stephanie.marchand@u-bordeaux2.fr
Analytica Chimica Acta
|January 28, 2010
Summary
A new HPLC method accurately measures gamma-glutamylcysteine and glutathione (GSH) in wine. It also quantifies oxidized glutathione (GSSG) for the first time, offering a validated assay for labs with limited equipment.
Area of Science:
- Analytical Chemistry
- Food Chemistry
- Biochemistry
Background:
- Glutathione (GSH) and its oxidized form (GSSG) are crucial antioxidants.
- Wine quality and health benefits are linked to its antioxidant content.
- Accurate quantification of these compounds in wine is challenging.
Purpose of the Study:
- To develop a simple, automated HPLC method for quantifying gamma-glutamylcysteine and reduced glutathione (GSH) in wines.
- To establish a method for quantifying oxidized glutathione (GSSG) in wines, a first for this matrix.
- To validate the developed method for laboratory use, including those with limited resources.
Main Methods:
- High-performance liquid chromatography (HPLC) with pre-column derivatization using 2,3-naphthalenedialdehyde (NDA).
- Isocratic separation enhanced by beta-cyclodextrine and fluorimetric detection.
- A novel pre-derivatization reduction step using glutathione reductase for GSSG quantification.
Main Results:
- Successful separation and quantification of gamma-glutamylcysteine and GSH.
- First-time quantification of GSSG in wine samples.
- The method demonstrated excellent sensitivity, selectivity, linearity, and accuracy.
- Validation confirmed reliability for low-resource laboratories.
Conclusions:
- A robust and validated HPLC method is available for comprehensive glutathione analysis in wine.
- This method facilitates better understanding of wine's antioxidant profile.
- The assay is suitable for routine quality control and research in diverse laboratory settings.
