Analysis of targeted phenolic ageing markers in Syrah red wines during bottle ageing: Influence of cork oxygen transfer rate.

A Syrah red wine ageing experiment was set up during 24-months and the influence of four micro-agglomerated corks were investigated. Specific phenolic ageing markers were selected and hemi-synthesized: vitisin B, malvidin-ethyl-catechin, and epicatechin-sulfonate. A targeted quantification method of these markers was then developed and validated by using ultra-high performance liquid chromatography - triple quadrupole mass spectrometry (UHPLC-QqQ-MS) operating in MRM (Multiple Reaction Monitoring). Results showed a significant decline in native grape polyphenol levels (anthocyanins, flavanols) as ageing progresses while pyranoanthocyanins, ethyl-linked pigments, and flavanol-sulfonates content increased. The cork oxygen transfer rate emerged as a pivotal factor and had significant effects on polyphenolic concentration evolution but had no significant impact on flavanol-sulfonate formation. These results provide valuable insights into the chemical evolution ongoing during wine ageing, accentuating the pivotal role of cork stopper selection in preserving wine quality over time.

Oxidation of Wine Polyphenols by Electrochemical Means in the Presence of Glutathione.

The oxidation of wine may be beneficial or harmful to its quality. On the one hand, controlled oxidation can lead to the development of desirable sensory characteristics for red wine, such as enhanced color stability. Alternatively, oxidation can lead to white wine browning and a decrease in fruity aromas, and the development of an off flavor and wine polyphenols are also involved. The presence of glutathione (GSH) can help mitigate the negative effects of oxidation by acting as a protective antioxidant. In order to better understand the antioxidant role played by GSH, wine polyphenols oxidation experiments by electrochemical means in the presence of GSH were carried out. The oxidation behavior of polyphenols representing different phenolic classes commonly found in wines, including protocatechuic acid (PCA), caffeic acid (CAF), epicatechin (EC), and rutin (Ru), was investigated using cyclic voltammetry and bulk electrolysis. We identified the oxidation products and reaction pathways of these polyphenols using ultra-high-performance liquid chromatography coupled with mass spectrometry (UPLC-MS), in both the absence and the presence of glutathione (GSH). UPLC-MS was utilized to demonstrate that, in the presence of glutathione (GSH), the four molecules were subjected to electrochemical oxidation, resulting in the formation of mono- and bi-glutathione conjugates. A two-electron oxidation process combined with the removal of two protons is the first step in transforming polyphenol molecules. As a result, the corresponding quinone is formed. The quinone can then be reduced back to its original form by glutathione (GSH), or it can interact further with GSH to produce mono- and bi-glutathione conjugates. These results contribute to understanding and predicting the oxidative degradation pathway of polyphenols in wine. Understanding this process seems important for winemakers to control and optimize the sensory characteristics of their wines.

Impact of phenolic composition and antioxidant parameters on the ageing potential of Syrah red wines measured by accelerated ageing tests.

Fourteen Syrah red wines with different initial composition and antioxidant properties (polyphenols, antioxidant capacity, voltammetric behaviour, colour parameters and SO2) were selected. Three different accelerated ageing tests (AATs) were then performed on these wines: thermal test at 60 °C (60 °C-ATT), enzymatic test with laccase (Laccase-ATT) and chemical test with H2O2 (H2O2-ATT). The results showed high correlations between the initial phenolic composition and antioxidant properties of the samples. Partial least squares (PLS) regressions were used in order to establish some models that can predict the AATs test results based on their different initial composition and antioxidant properties. The PLS regression models had overall very good accuracy and involved different explaining variables for each test. The models taking into account all the measured parameters and the phenolic composition alone showed good predictive capacities with correlation coefficients (r2) > 0.89.

Impact of Acetaldehyde Addition on the Sensory Perception of Syrah Red Wines.

Two experimental Syrah red wines with different polyphenol contents were used to study the impact of acetaldehyde addition on olfactory perception. Free acetaldehyde levels were measured in red wine by Head Space-Gas Chromatography-Mass Spectrometry (HS-GC-MS) to determine the acetaldehyde combination levels for those wines. Significant differences were observed for both sensory threshold and acetaldehyde combination for the wines. A descriptive sensory analysis of the wines was then performed by using a trained panel and a Hierarchical-Check-All-That-Apply (HCATA) analysis of the wines with or without acetaldehyde addition. The results show that classical cited sensory descriptors for acetaldehyde (overripe apple and oxidized apple) varied significantly between the control wines and those with acetaldehyde addition. Non-acetaldehyde related descriptors (fresh vegetable, fresh flowers, cocoa, and meat juice) were also significantly impacted in the samples with increasing acetaldehyde additions. This suggests possible interactions between acetaldehyde and other volatile compounds that can create antagonistic or synergistic effects between the molecules or at the olfactory receptor level.

UHPLC-Q-Orbitrap /MS2 identification of (+)-Catechin oxidation reaction dimeric products in red wines and grape seed extracts.

B-type procyanidin dimers and (+)-catechin dimeric oxidation products were analyzed in grape seed extracts and red wines (UHPLC-Q-Orbitrap MS). The different dimers had different fragmentation patterns according to their interflavan linkage position. Oxidation dimeric compounds had a specific fragment ion at m/z 393, missing for B-Type dimers fragmentations. A fragment ion at m/z 291 occurred and was specific for oxidation dimeric compounds with a COC linkage. Higher level oxidation products had abundant specific fragments: m/z 425, 397 and 245. These fragmentations were useful to identify them in complex samples such as grape seed extracts and wines. Three grape varieties and three ripening stages were selected and the corresponding seed extracts were obtained. The analyses revealed an increasing trend for the oxidation markers during grape ripening. The analysis of Syrah wines (2018, 2014, 2010) showed a decreasing trend of these molecules during wine ageing which might be due to further oxidation.

Study of the oxidative evolution of tannins during Syrah red wines ageing by tandem mass spectrometry.

Red wine is a very complex medium in which condensed tannins undergo many modifications during winemaking and bottle ageing. These reactions have an impact on the organoleptic properties. This work aimed to highlight tannins evolution related to wine evolution by studying three vintages of Syrah wines. An accelerated oxidation was also undertaken in order to evaluate the ability of this oxidation to imitate natural evolution. After chemical depolymerization of the tannins, the monitoring of 6 types of markers at two oxidation levels was investigated. An evolution of the tannin oxidation state during ageing evidenced by the increase of the markers of the second oxidation level was observed. In the 2018 oxidized wine sample, the first oxidation level markers were similar to the 2014 vintage but the second oxidation level markers were higher than other vintages, indicating a more advanced state of tannin oxidation.

Identification of adducts between oxidized rosmarinic acid and glutathione compounds by electrochemistry, liquid chromatography and mass spectrometry.

Natural polyphenols are omnipresent and are an integral part of the human diet as well as quinones. Glutathione (GSH) is present in a significant amount inside cells and consequently, GSH conjugates of polyphenols will be encountered in the body. In the current work, voltammetry and liquid chromatography-mass spectrometry were carried out to characterize the reaction mechanism of the electrochemical oxidation of polyphenolic rosmarinic acid (RA) with GSH nucleophiles in aqueous solution. Electrochemical investigation of RA revealed that two consecutive transfer steps (which depend on pH) of two electrons and protons occur during the reversible oxidation of RA. Moreover, it was found that the first oxidation step is associated with the 3,4-dihydroxyphenyl lactic acid moiety, whereas the second oxidation step corresponds to the oxidation of the caffeic acid one. By using ultrahigh-performance liquid chromatography-diode array detection-mass spectrometry (UPLC-DAD-MS) in the negative ion mode, the oxidation pathways of RA in the presence of GSH were analyzed, and a total of four RA-GSH conjugates were identified. The oxidative degradation pathway of RA can be better apprehended and forecasted by the acquired results in this study.

Red Wine Oxidation Characterization by Accelerated Ageing Tests and Cyclic Voltammetry.

In order to obtain information on the oxidative behavior of red wines, oxygen consumption rates and electrochemical changes (cyclic voltammetry) were measured for nine red wines subject to three different accelerated ageing tests: chemical (with hydrogen peroxide), enzymatic (with laccase from Trametes versicolor), and temperature (at 60 °C). Oxidative behavior depended both on the wine sample and accelerated ageing test type. A good correlation was observed between electrochemical parameters of charges for reference/non-oxidized wines, in accordance with their antioxidant capacity, and the variation of charges after enzymatic and temperature tests, meaning that cyclic voltammetry could be used in order to predict these two oxidation tests and reflect the wine sensitivity towards respective oxidation targets. However, it was not possible to predict wine chemical oxidation test based on hydrogen peroxide from the electrochemical measurements.

Unambiguous NMR Structural Determination of (+)-Catechin-Laccase Dimeric Reaction Products as Potential Markers of Grape and Wine Oxidation.

(+)-Catechin-laccase oxidation dimeric standards were hemi-synthesized using laccase from Trametes versicolor in a water-ethanol solution at pH 3.6. Eight fractions corresponding to eight potential oxidation dimeric products were detected. The fractions profiles were compared with profiles obtained with two other oxidoreductases: polyphenoloxidase extracted from grapes and laccase from Botrytis cinerea. The profiles were very similar, although some minor differences suggested possible dissimilarities in the reactivity of these enzymes. Five fractions were then isolated and analyzed by 1D and 2D NMR spectroscopy. The addition of traces of cadmium nitrate in the samples solubilized in acetone-d6 led to fully resolved NMR signals of phenolic protons, allowing the unambiguous structural determination of six reaction products, one of the fractions containing two enantiomers. These products can further be used as oxidation markers to investigate their presence and evolution in wine during winemaking and wine ageing.

Polyphenolic Characterization of Merlot, Tannat and Syrah Skin Extracts at Different Degrees of Maturity and Anti-Inflammatory Potential in RAW 264.7 Cells.

(1) Background: Both sensory quality and healthy attributes of Vitis vinifera grapes used for winemaking are closely related with the polyphenolic composition of their skins. (2) Methods: In this study, the polyphenolic characterization (flavan-3-ols, procyanidins, flavonols, stilbenes, anthocyanins) was investigated by ultra performance liquid chromatography coupled to a triple quadrupole mass spectrometer (UPLC-QqQ-MS). Skins from Vitis vinifera Merlot, Tannat, and Syrah red grape varieties cultivated in the south of France at different stages of ripening in 2018 were used. The anti-inflammatory and the antioxidant potential of the extracts were evaluated by the measure of nitric oxide (NO) and the intracellular reactive oxygen species production (ROS) in lipopolysaccharide (LPS)-stimulated macrophages. (3) Results: 41 polyphenols were quantified in all samples. Generally, the flavan-3-ol and procyanidin content decreased during ripening whereas the anthocyanins and stilbenes increased. In addition, as a novelty of this work, a wide identification and characterization of monomeric and oligomeric stilbenes was assessed by using authentic standards isolated in our laboratory, some of them (parthenocissin A and miyabenol C) reported for the first time in Merlot, Tannat and Syrah cultivars. The before-veraison skin extracts of all studied varieties, exhibited higher NO and ROS productions inhibition (>50%) proving both antioxidant and anti-inflammatory properties.

Voltammetric Behavior, Flavanol and Anthocyanin Contents, and Antioxidant Capacity of Grape Skins and Seeds during Ripening (Vitis vinifera var. Merlot, Tannat, and Syrah).

Skin and seed grape extracts of three red varieties (Merlot, Tannat, and Syrah) at different stages of ripening were studied for their total phenolic content (TPC) by using the Folin-Ciocalteu assay and for their total antioxidant capacity (TAC) by using spectrophotometric and electrochemical assays. Flavanol and anthocyanin compositions were also investigated using Ultra Performance Liquid Chromatography coupled with Mass Spectrometry (UPLC-MS). Results showed that seeds had the highest phenolic content and the highest antioxidant potential compared to skins at all stages of ripening. The highest TPC and TAC values were measured in seeds at close to veraison and veraison ripening stages. In skins, the highest values were found at the green stage, it was in accordance with the flavanols content. The voltammetric measurements were carried out using disposable single walled carbon nanotubes modified screen-printed carbon electrodes (SWCNT-SPCE). Three peaks on voltammograms were obtained at different oxidation potentials. The first anodic peak that oxidized at a low potential describes the oxidation of ortho-dihydroxy phenols and gallate groups, the second peak corresponds to the malvidin anthocyanins oxidation and the second oxidation of flavonoids. The third voltammetric peak could be due to phenolic acids such as p-coumaric acid and ferulic acid or the second oxidation of malvidin anthocyanins. The high linear correlation was observed between antioxidant tests and flavanols in skins (0.86 ≤ r ≤ 0.94), while in seeds, 'r' was higher between electrochemical parameters and flavanols (0.64 ≤ r ≤ 0.8).

Red Wine Oxidation: Accelerated Ageing Tests, Possible Reaction Mechanisms and Application to Syrah Red Wines.

Wine oxidation and ageing involve many complex chemical pathways and reaction mechanisms. The purpose of this study is to set up new and reproducible accelerated red wine ageing tests and identify chemical oxidation or ageing molecular markers. Three accelerated and reproducible ageing tests were developed: a heat test (60 °C); an enzymatic test (laccase test; a chemical test (hydrogen peroxide test). Depending on the test, oxygen consumption was significantly different. For a young wine (2018), the oxygen consumption rate moved from 2.40 ppm.h-1 for the heat test to 3.33 ppm.h-1 for the enzymatic test and 2.86 ppm.h-1 for the chemical test. Once applied to two other vintages (2010 and 2014) from the same winery, the tests revealed different comportments corresponding to wine natural evolution. High resolution UPLC-MS was performed on forced ageing samples and compared to naturally aged red wines. Specific oxidation or ageing ion markers were found with significant differences between tests, revealing the specificity of each test and different possible molecular pathways involved. The hydrogen peroxide test seems to be closer to natural oxidation with an important decrease in absorbance at 520 nm and similar molecular ion variations for [M+H]+ = 291, 331, 347, 493, 535, 581, 639 Da.

Multimethod Approach for Extensive Characterization of Gallnut Tannin Extracts.

Gallotannins extracted from gallnuts are commonly added to wine to improve its properties. They consist of mixtures of galloylester derivatives of glucose. However, their composition and properties are not well-established. In this study, methods based on liquid chromatography coupled to ultraviolet-visible detection and mass spectrometry, size-exclusion chromatography, and one-dimensional (31P) and two-dimensional (1H diffusion ordered spectroscopy, 31P total correlated spectroscopy, and 1H/13C heteronuclear single-quantum correlation and heteronuclear multiple-bond correlation) nuclear magnetic resonance spectroscopies have been implemented for extensive chemical characterization of three commercial gallnut tannin extracts. Differences in the proportions of the different constituents (gallic, digallic, and trigallic acids and galloylglucose derivatives) and in the structure and molecular weight distributions of gallotannins were demonstrated between the three extracts, with chains containing 8.5, 12.2, and 12.4 galloyl groups on average for TAN A, TAN B1, and TAN B2, respectively. The antioxidant capacities of the extracts, evaluated using the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) method, were similar and related mostly to their total tannin content, with only a limited impact of the tannin composition.

[Estimation of urea distribution volume in hemodialysis patients]. / Estimation du volume de distribution de l'urée chez le patient hémodialysé.

BACKGROUND: On-line urea clearance estimation, currently available on some dialysis monitors, makes it possible to calculate the dialysis dose Kt and thus allows to estimate Kt/V for each session, providing an estimation of urea distribution volume (V) at equilibrium assumed equal to total body water. METHODS: Three methods suitable for routinely estimating V, using the anthropometric Watson formula (V(Wat)), the body composition monitor (BCM) device (Fresenius Medical Care) based on bio-impedance analysis (V(imp)) and the indirect estimation (V(Daug)) obtained from measurement of Kt/(Kt/V)(sp) ratio respectively are compared during 25 dialysis sessions in 15 patients to a direct estimation (V(DDQ)) obtained by direct quantification of dialysis (DDQ) considered as the gold standard in hemodialysis patient.. RESULTS: V(Watson) overestimates V(DDQ) by about 20%. The values of V(imp) (29.1±5.6 L) and V(Daug) (29.5±4.6 L) are in agreement with V(DDQ) (29.9±5.2 L). Correlation coefficient with V(DDQ) is better for V(imp) (r=0.94) than for V(Daug) (r=0.85). CONCLUSION: Bio-impedancemetry using BCM and indirect method using the second generation Daugirdas equation are two methods of clinical interest for estimating V. Bio-impedancemetry does not require blood sample, but it needs to have a specific device at disposal.