 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

ANALYZING THE ROLE OF ELEMENTAL SULFUR IN GRAPE JUICE ON THE DEVELOPMENT OF POLYFUNCTIONAL MERCAPTANS IN SAUVIGNON BLANC WINES

Sauvignon blanc is characterized by distinctive aromas, both fruity and herbaceous. The “green” character has been attributed to the methoxypyrazines, while the “fruity” character is associated with polyfunctional mercaptans . Polyfunctional mercaptans are of great significance due to their high impact on wines and associated low perception thresholds.
Elemental sulfur (S⁰) is widely used to protect grapevines from powdery mildew.

Cascading effects of spring weather conditions into grape berry ripening

The effects of climate change on viticulture are complex due to interactions among factors and cascading effects.

Apoplastic pH influences Vitis vinifera Barbera recovery responses to short and prolonged drought

Alteration of sap pH is one of the first chemical changes that occurs within the xylem vessels of plants exposed to drought. Xylem sap acidification accompanied by the accumulation of soluble sugars has been recently documented in several species (Sharp and Davis, 2009; Secchi and Zwieniecki, 2016). Here, Vitis vinifera plants of the anysohydric cultivar Barbera were exposed to either short (no irrigation; SD) or to prolonged drought (continual reduction of 10% water; PD). When comparable severe stress was reached, the potted grapes were re-watered. SD was characterized by fast (2–3 days) stomatal closure and high abscisic acid (ABA) accumulation in xylem sap (>400 μg L−1) and in leaf. In PD plants, the rise in ABA levels was considerably diminished.

Antioxidant activity of grape seed and skin extract during ripening

Reactive oxygen species (ROS) play an important physiological role in the body’s defense and being involved in numerous signaling pathways 1, 2. When the balance between oxidant and antioxidant species is altered in favor of ROS, oxidative stress is generated. In this condition the cells are damaged as the ROS oxidize important cellular components, such as proteins, lipids, nucleic acids and

The moment of preharvest elicitor application influence its final effect on winegrapes quality

Phenolic compounds are secondary metabolites of grapes. Plants produce a wide variety of this type of metabolites through diverse biosynthesis pathways and their production is sometimes a response to external stimuli, either environmental or biotic stresses. Some of them may act as chemical defenses against pathogens or herbivores and their synthesis is increased when the attack exists. However, it is remarkable that the synthesis of these interesting compounds can be activated even when the stimulus is not present, with the use of elicitors. These are substances that when applied exogenously trigger the biosynthetic pathways conducting to the synthesis of these defense compounds.