 IVES 9 IVES Conference Series 9 Evaluating analytical methods for quantification of glutathione in grape juice and wine

Evaluating analytical methods for quantification of glutathione in grape juice and wine

Abstract

AIM: Glutathione (GSH) is a powerful natural antioxidant, considered as a promising molecule against oxidative damage of aroma during winemaking and storage. His concentration in the grape juice is variable. Enological practice in the cellar can promote the preservation of the GSH in the grape juice. Recently the prescription of OIV allows to add glutathione rich substances to the must. This new practice creates an increasing interest in the quantification of GSH. Several analytical methods were published to measure GSH[1,2,3,4] and his dimer (GSSG) separately[5] or together as total glutathione[6] content. In this work we compared two analytical methods for the analyses of grape juice and wine samples.

METHODS: The first method is an enzymatic assay (EA), based on the reaction of thiol with DTNB in the presence of glutathione reductase enzyme to measure the total glutathione content. This method was automatized to allow high through-put measurements in the concentration range of 5-100mg/l. The second method, using UPLC-MS/MS, is more sensitive (LOD = 0.5mg/l) and permits simultaneous quantification of GSH, GSSG and additionally the sulfonated form of glutathione (GSSO3H).

RESULTS: The best results were obtained with 2.5g/l ascorbic acid. Using the two analytical methods, we found a strong correlation (R2=0.98) between the total glutathione (EA) and the sum of GSH and GSSG (UPLC-MS/MS) in grape juice samples (n>100), where the GSSO3H concentration was low (0-8 mg/l) comparing to the GSH and GSSG (5-100mg/l). In wine samples the total glutathione concentration was low (2-7mg/l) and the GSSO3H was more important (5-9mg/l) due to the combination of SO2 with the glutathione. View the high reactivity of GSH, a special attention should be accorded to the preparation and the storage of grape juice samples. We compared the effect of different concentration of SO2 and ascorbic acid as additives and found that 2.5g/l ascorbic acid gave the best results.

CONCLUSION

Based on our results the enzymatic assay is an economic alternative to measure the total glutathione concentration of grape juice. However for wine the UPLC-MS/MS method is recommended, to reach the necessary sensitivity and to analyze all glutathione species.

DOI:

Publication date: September 7, 2021

Issue: Macrowine 2021

Type: Article

Authors

Ágnes Dienes-Nagy

Agroscope, Nyon, Switzerland,Frédéric VUICHARD, Agroscope, Nyon, Switzerland Marie BLACKFORD, Agroscope, Nyon, Switzerland Fabrice LORENZINI, Agroscope, Nyon, Switzerland

Contact the author

Keywords

glutathione, enzymatic assay, uplc-ms/ms

Citation

Response to powdery and downy mildew of varieties with disease resistance genes (PIWI)

Erysiphe necator and Plasmopara viticola are the causal agents of powdery and downy mildew on grapevines, leading to significant economic losses. Numerous chemical treatments are applied to control these diseases, leading to environmental problems and the appearance of resistance to these products. Therefore, the study of new strategies to achieve the objectives of sustainable development is a priority. In this sense, the use of new varieties resistant to these diseases may be an option of interest. The objective of this work was to analyze the degree of resistance of 9 varieties with downy mildew resistance genes (Rpv3 and/or Rpv12), four of which also carry a powdery mildew resistance gene (Ren 1) by in vitro inoculation assays.

Zoning, environment, and landscape: historic and perspective

Dans une approche globale, nous proposons la définition suivante du zonage : “représentation cartographique associée à une sectorisation du territoire en zones unitaires homogènes à partir de facteurs discriminants établis sur la base d’indicateurs quantifiables et d’avis d’experts”. La première application de cette méthode a porté sur la caractérisation du terroir en liaison avec les aspects qualitatifs des vins. Il est également possible d’envisager d’appliquer cette démarche dans les stratégies environnementales et paysagères liées aux approches territoriales et aux pratiques viticoles. Cette méthode peut servir de base dans la mise en œuvre des outils financiers associés aux mesures environnementales (CTE, aides spécifiques).

Maturation of Agiorgitiko (Vitis vinifera) red wine on its wine lees: Impact on its phenolic composition

Maturation of wine on lees (often referred as sur lie) is a common practice applied by many winemakers around the world. In the past this method was applied mainly on white and/or sparkling wine production but recently also to red wine production. In our experiment, we matured red wine on wine lees of two origins: a) Light wine lees, collected after the completion of the alcoholic fermentation, b) Heavy lees, collected after the completion of the malolactic fermentation. The lees were free of off-odors and were added in the red wine in percentage 3% and 8%, simulating common winemaking addition. The maturation lasted in total six months and samples were collected for analysis after one, three and six months. During storage the lees were stirred.

Measuring elemental sulfur in grape juice in relation to varietal thiol formation in Sauvignon blanc wines.

Aim: Sauvignon blanc displays a range of styles that can include prominent tropical and passionfruit aromas. Both sensory evaluation and chemical analysis have confirmed the above-average presence of ‘varietal thiols’ in the Sauvignon blanc wines from Marlborough, New Zealand.

First disclosure of eugenol precursors in Vitis genus: analytical development and quantification

The main aim of this work was to develop an analytical method to disclosure the
molecular form of eugenol precursor. Indeed eugenol is an important contributor to
Armagnac spirits typicity made with Baco blanc.