Macrowine 2021
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

Related articles…

Towards a regional mapping of vine water status based on crowdsourcing observations

Monitoring vine water status is a major challenge for vineyard management because it influences both yield and harvest quality. It is also a challenge at the territorial scale for identifying periods of high water restriction or zones regularly impacted by water stress. This information is of major importance for defining collective strategies, anticipating harvest logistic or applying for irrigation authorisation. At this spatial scale, existing tools and methods for monitoring vine water status are few and often require strong assumptions (e.g. water balance model). This paper proposes to consider a collaborative collection of observations by winegrowers and wine industry stakeholders (crowdsourcing) as an interesting alternative. Indeed, it allows the collection of a large number of field observations while pooling the collection effort. However, the feasibility of such a project and its interest in monitoring vine water status at regional scale has never been tested.

The objective of this article is to explore the possibility of making a regional map of vine water status based on crowdsourcing observations. It is based on the study of the free mobile application ApeX-Vigne, which allows the collection of observations about vine shoot growth. This information is easy to collect and can be considered, under certain conditions, as a proxy for vine water status. This article presents the first results obtained from the nearly 18,000 observations collected by winegrowers and wine industry stakeholders during 2019, 2020 and 2021 seasons. It presents the vine shoot growth maps obtained at regional scale and their evolution over the three vintages studied. It also proposes an analysis of the factors that favoured the number of observations collected and those that favoured their quality. These results open up new perspectives for monitoring vine water status at a regional scale but above they provide references for other crowdsourcing projects in viticulture.

Long-term drought resilience of traditional red grapevine varieties from a semi-arid region

In recent decades, the scarcity of water resources in agriculture in certain areas has been aggravated by climate change, which has caused an increase in temperatures, changes in rainfall patterns, as well as an increase in the frequency of extreme phenomena such as droughts and heat waves. Although the vine is considered a drought-tolerant specie, it has to satisfy important water requirements to complete its cycle, which coincides with the hottest and driest months. Achieving sustainable viticulture in this scenario requires high levels of efficiency in the use of water, a scarce resource whose use is expected to be severely restricted in the near future. In this regard, the use of drought-tolerant varieties that are able to maintain grape yield and quality could be an effective strategy to face this change. During three consecutive seasons (2018-2020) the behavior in rainfed regime of 13 traditional red grapevine varieties of the Spain central region was studied. These varieties were cultivated in a collection at Centro de Investigación de la Vid y el Vino de Castilla-La Mancha (IVICAM-IRIAF) located in Tomelloso (Castilla-La Mancha, Spain). Yield components (yield, mean bunch and berry weight, pruning weight), physicochemical parameters of the musts (brix degree, total acidity, pH) and some physiological parameters related with water stress during ripening period (δ13C, δ18O) were analysed. The application of different statistical techniques to the results showed the existence of significant differences between varieties in their response to stressful conditions. A few varieties highlighted for their high ability to adapt to drought, being able to maintain high yields due to their efficiency in the use of water. In addition, it was possible quantify to what extent climate can be a determinant in the δ18O of musts under severe water stress conditions.

Grape berry size is a key factor in determining New Zealand Pinot noir wine composition

Making high quality but affordable Pinot noir (PN) wine is challenging in most terroirs and New Zealand’s (NZ) situation is no exception. To increase the probability of making highly typical PN wines producers choose to grow grapes in cool climates on lower fertility soils while adopting labour intensive practices. Stringent yield targets and higher input costs necessarily mean that PN wine cost is high, and profitability lower, in line-priced varietal wine ranges. To understand the reasons why higher yielding vines are perceived to produce wines of lower quality we have undertaken an extensive study of PN in NZ. Since 2018, we established a network of twelve trial sites in three NZ regions to find individual vines that produced acceptable commercial yields (above 2.5kg per vine) and wines of composition comparable to “Icon” labels. Approximately 20% of 660 grape lots (N = 135) were selected from within a narrow juice Total Soluble Solids (TSS) range and made into single vine wines under controlled conditions. Principal Component Analysis of the vine, berry, juice and wine parameters from three vintages found grape berry mass to be most effective clustering variable. As berry mass category decreased there was a systematic increase in the probability of higher berry red colour and total phenolics with a parallel increase in wine phenolics, changed aroma fraction and decreased juice amino acids. The influence of berry size on wine composition would appear stronger than the individual effects of vintage, region, vineyard or vine yield. Our observations support the hypothesis that it is possible to produce PN wines that fall within an “Icon” benchmark composition range at yields above 2.5kg per vine provided that the Leaf Area:Fruit Weight ratio is above 12cm2 per g, mean berry mass is below 1.2g and juice TSS is above 22°Brix.

Phenological characterization of a wide range of Vitis Vinifera varieties

In order to study the impact of climate change on Bordeaux grape varieties and to assess the adaptation capacities of candidates to the grape varieties of this wine region to the new climatic conditions, an experimental block design composed of 52 grape varieties was set up in 2009 at the INRAE Bordeaux Aquitaine center. Among the many parameters studied, the three main phenological stages of the vine (budburst, flowering and veraison) have been closely monitored since 2012. Observations for each year, stage and variety were carried out on four independent replicates. Precocity indices have been calculated from the data obtained over the 2012-2021 period (Barbeau et al. 1998). This work allowed to group the phenological behaviour of the grapevine varieties, not only based on the timing of the subsequent developmental stages, but also on the overall precocity of the cycle and the total length of the cycle between budburst and veraison. Results regarding the variability observed among the different grape varieties for these phenological stages are presented as heat maps.

Biodiversity in the vineyard agroecosystem: exploring systemic approaches

Biodiversity conservation and restoration are essential for guarantee the provision of ecosystem services associated to vineyard agroecosystem such as climate regulation trough carbon sequestration and control of pests and diseases. Most of published research dealing with the complexity of the vineyard agroecosystems emphasizes the necessity of innovative approaches, including the integration of information at different temporal and spatial scales and development of systemic analysis based on modelling. A biodiversity survey was conducted in the Franciacorta wine-growing area (Lombardy, Italy), one of the most important Italian wine-growing regions for sparkling wine production, considering a portion of the territory of 112 ha. The area was divided into several Environmental Units (EUs), defined as a whole vineyard or portion of vineyard homogenous in terms of four agronomic characteristics: planting year, planting density, cultivar, and training system. In each EU a set of compartments was identified and characterised by specific variables. The compartments are meteorology, morphology (altitude, slope, aspect, row orientation, and solar irradiance), ecological infrastructures and management. The landscape surrounding EU was also characterised in terms of land-use in a buffer zone of 500 m. For each component a specific methodology was identified and applied. Different statistical approaches were used to evaluate the method to integrate the information related to different compartments within the EU and related to the buffer zone. These approaches were also preliminarily evaluated for their ability to describe the contribution of biodiversity and landscape components to ecosystem services. This methodological exploration provides useful indication for the development of a fully systemic approach to structural and functional biodiversity in vineyard agroecosystems, contributing to promote a multifunctional perspective for the all wine-growing sector.