Red Wine Phenolics Research Articles

Inhibition of ALDH2 by quercetin glucuronide suggests a new hypothesis to explain red wine headaches

The consumption of red wine induces headaches in some subjects who can drink other alcoholic beverages without suffering. The cause for this effect has been attributed to a number of components, often the high level of phenolics in red wine, but a mechanism has been elusive. Some alcohol consumers exhibit flushing and experience headaches, and this is attributed to a dysfunctional ALDH2 variant, the enzyme that metabolizes acetaldehyde, allowing it to accumulate. Red wine contains much higher levels of quercetin and its glycosides than white wine or other alcoholic beverages. We show that quercetin-3-glucuronide, a typical circulating quercetin metabolite, inhibits ALDH2 with an IC50 of 9.6 µM. Consumption of red wine has been reported to result in comparable levels in circulation. Thus, we propose that quercetin-3-glucoronide, derived from the various forms of quercetin in red wines inhibits ALDH2, resulting in elevated acetaldehyde levels, and the subsequent appearance of headaches in susceptible subjects. Human-subject testing is needed to test this hypothesis.

White wine phenolics: current methods of analysis.

White wine phenolic analyses are less common in the literature than analyses of red wine phenolics. Analytical techniques for white wine phenolic analyses using spectrophotometric, chromatographic, spectroscopic, and electrochemical methods are reported. The interest of research in this area combined with the advances in technology aimed at the winemaking industry are promoting the establishment of novel approaches for identifying, quantifying, and classifying phenolic compounds in white wine. This review article provides an overview of the current research into white wine phenolics through a critical discussion of the analytical methods employed. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effects of grape phenolics, myricetin and piceatannol, on bovine granulosa and theca cell proliferation and steroid production in vitro

Myricetin (a flavonol) and piceatannol (a stilbenoid) are naturally occurring phenolic compounds in red wine with cardio-protective and anti-carcinogenic effects, but their potential reproductive effects have not been investigated. Thus, the present study was designed to determine if myricetin and piceatannol can directly affect ovarian function using bovine granulosa cells (GC) and theca cells (TC) as in vitro model systems to evaluate effects on cell proliferation and steroid production. In Experiment 1 and 2, myricetin and piceatannol at 30 μM blocked insulin-like growth factor 1 (IGF1)-induced progesterone production by GC without affecting GC numbers. In contrast, myricetin stimulated IGF1-induced estradiol production, whereas piceatannol at 30 μM inhibited IGF1-induced estradiol production by 90% in GC. In Experiment 3 and 4, TC androstenedione and progesterone production and TC proliferation was inhibited by myricetin and piceatannol at 30 μM. In Experiment 5, piceatannol (30 μM) reduced the Fusarium mycotoxin, beauvericin (6 μM)-induced inhibition on progesterone production and cell proliferation. Myricetin (30 μM) reduced the inhibitory effect of beauvericin on estradiol but not progesterone production or cell proliferation. In conclusion, the red wine phenols, myricetin and piceatannol, directly affected GC and TC steroidogenesis, and were able to reduce some of the inhibitory effects of beauvericin on GC function.

The macromolecular diversity of Italian monovarietal red wines

While red wine phenolics have been extensively studied, polysaccharides and proteins have not received the same level of attention, especially when considering Italian wines. In this study, for the first time, quantitative and qualitative data on the macromolecular (proteins and polysaccharides) and tannin composition of 110 monovarietal red wines from 11 of the most important Italian grape varieties are reported. The winemaking did not include any filtration, oak contact, fining treatments, or ageing on yeast lees. Results highlighted a great inter- and intra- varietal diversity. The protein content ranged between 0 and 159 mg/L, polysaccharides between 211 and 1081 mg/L and total tannins between 171 and 3746 mg/L, with averages of 41 mg/L, 497 mg/L and 1687 mg/L, respectively. Six varieties with protein content representative of the variability observed were selected and submitted to electrophoresis. Within each variety, the SDS-PAGE mobility of protein-tannin complexes was similar but showed two distinct patterns for wines of different varieties (higher mobility for Corvina, Teroldego and Raboso Piave, lower mobility for Nebbiolo, Aglianico, Cannonau), suggesting that the Italian monovarietal wines can be diverse also in their colloidal-forming structures. This can be explained by looking at the different percentages of protein-reactive tannins (TBSA) on the total tannin content (TMCP), which is a varietal characteristic.

Abundant Existence of Bovine Serum Albumin Non-precipitable Tannins in Red Wines

Phenolics in red wines made from three cultivars, Muscat Bailey A (MBA), Merlot (MR), and Cabernet Sauvignon (CS), were separated into four fractions using the bovine serum albumin (BSA) precipitat...

Direct quantification of red wine phenolics using fluorescence spectroscopy with chemometrics

Phenolic compounds are secondary metabolites known to play crucial roles in important chemical reactions impacting the mouthfeel, colour and ageing potential of red wine. Their complexity has resulted in a number of advanced analytical methods, which often prevent routine phenolic analysis in winemaking. Fluorescence spectroscopy could be an alternative to current spectrophotometric techniques and its combination with chemometrics was investigated for its suitability in directly quantifying phenolic content of unaltered red wine and fermenting samples. Front-face fluorescence was optimised and used to build predictive models for total phenols, total condensed tannins, total anthocyanins, colour density and polymeric pigments. Machine learning algorithms were used for model development. The most successful models were built for total phenols, total condensed tannins and total anthocyanins with coefficient of determination (R2cal) and RMSECV of 0.81, 0.89, 0.80 and 5.71, 104.03 mg/L, 60.67 mg/L, respectively. The validation results showed R2val values of 0.77, 0.8 and 0.77, and RMSEP values of 7.6, 172.37 mg/L and 76.57 mg/L, respectively. A novel approach for the classification of South African red wine cultivars based on unique fluorescent fingerprints was also successful with an overall cross validation score of 0.8. The best classification ability (validation score = 0.93) was shown for the data set containing only fermenting wines for the most widely represented cultivars (>20 samples). This approach may provide a useful tool for authentication and quality control by regulatory bodies.

How acetaldehyde reacts with low molecular weight phenolics in white and red wines

Acetaldehyde is a key compound in determining wine color evolution and sensory properties. Major wine metabolites reactive to acetaldehyde are phenolic compounds, mainly flavan-3-ols and anthocyanins. Many studies have been conducted with the purpose of investigating acetaldehyde reactivity in model solutions, but very poor are the reports of its fate in real wines. By means of LC-HRESIMS and UV/Vis HPLC, red and white wines exposed to an excess of acetaldehyde were analyzed with a specific focus on low molecular weight phenolics. The chemical behavior of acetaldehyde turned out to be different in white and red wines. In white wines, it mainly mediated the formation of vinyl-flavan-3-ol derivatives, while in red wines it led to the formation of ethylidene-bridged red pigments. These latter positively enhanced the color properties of red wines. Conversely, in white wines, the formation of compounds, such as xanthylium ions, causing the undesired browning effects were not detected.

Mathematical modelling to enhance winemaking efficiency: a review of red wine colour and polyphenol extraction and evolution

Red wine quality is determined greatly by phenolic substances whose presence relies on extraction of grape skins and seeds during fermentation. Monitoring phenolic behaviour is beneficial for fermentation management because of the importance of maceration during red winemaking and its influence on phenolics that contribute to predominant red wine characteristics, such as colour, flavour and mouthfeel. Properly quantifying the kinetics associated, however, with red grape and wine phenolics is an intricate process. This review focuses on the extraction and ensuing reactions of principal phenolic substances during various stages of red winemaking as described by dynamic and spatial mathematical models. The extraction and reaction phenomena are concurrently influenced by several environmental and technological factors during fermentation, such as temperature, oxidation and mixing regimes, resulting in complex phenolic behaviour over time. As such, employing mathematical modelling is extremely useful for ascertaining the kinetic and spatial basis for phenolic behaviour, and the models significantly enhance the existing understanding of the extraction and reaction mechanisms of phenolic substances that are critical to red wine quality. Ultimately, this knowledge will allow for the development of predictive tools for phenolic extraction based on fruit composition and can potentially contribute to increased wine quality with optimised and sustainable production practices.

How must pH affects the level of red wine phenols

Slight variations of pH affect the sensory properties and stability of wines with important implications for the winemaking practices. Preliminary studies in wine-like solutions with pH values ranging from 3 to 4 showed that anthocyanin solubility decreased as the pH increased. A significant drop of solubility was observed at pH 3.3. In order to verify these results in real wines, an experimental protocol was designed with the purpose of investigating the pH influence on the anthocyanin extractability in three musts differing from each other only by the pH levels adjusted to 3.2, 3.5 and 3.9, respectively. The NMR-based analysis of the wines, deriving from the three musts, provided data consistent with those observed in wine-like solutions. The extraction of anthocyanins turned out to be increasingly lower as the pH increased even if the effect of pH was more contained than expected on the basis of the data obtained from model solutions. Other molecules of enological interest, mainly catechins and cinnamates, were proven to be affected by the must pH. Additionally, tannins slightly increased as the pH increased but the reactivity of tannins towards saliva decreased confirming the decisive role of pH also on wine astringency.

Processes and purposes of extraction of grape components during winemaking: current state and perspectives.

The quality of red wine is dependent on the presence not only of volatile compounds that influence wine aroma but also on phenolic compounds due to their impact on mouthfeel, colour, flavour and ageing potential. Whereas wine aroma arises from the grapes, action of microorganisms and maturation phenomena, red wine phenolics are predominantly found in the grape berry skins and seeds and are extracted during maceration into the fermenting must (i.e. grape solids and juice). Thus, although every step in winemaking has a significant impact on the outcome of the finished product, it can be considered that fermentation and the ensuing extraction of grape components are the most critical value adding step in the process. This mini-review examines such aspects, and against this backdrop, considers the need for improvements to fermentation control during wine production, to optimise the outcome of extraction from grape solids. Specifically, there is interest in modulating and forecasting wine composition based on grape specifications, and with this, the ability to regulate winery production practices to achieve target quality specifications. Such predictions and measures are proposed to help winemakers adapt to emerging issues associated with climate change. KEY POINTS: • Definition and description of important grape metabolites extracted during winemaking. • Description of processes influencing development and extraction of grape components. • Includes key aspects related to vineyard, harvesting, winemaking, and ageing. • Covers future trends regarding extrac Mini-Reviewtion, climate change and production efficiency.

Modification of the organic acid profile of grapes due to climate changes alters the stability of red wine phenolics during controlled oxidation

The effect of the main grape organic acids (tartaric, malic and citric) on the degradative oxidation of red wine was investigated by NMR, HPLC and spectrophotometry. Wines featuring the same pH value of 3.2 with different combinations of organic acids were prepared. Results showed that tartaric acid preserved native anthocyanins from oxidative degradation more than malic and citric acids, with malic acid being the one favoring oxidations the most and, consequently, acetaldehyde production. Wines richer in malic acids showed the highest reactivity towards saliva proteins and a potential higher astringency. Given the wide changes in tartaric/malic acid ratio with climate, these results can help to act in vineyard, as well as in winery, to manage the malic/tartaric acid ratio with the aim of improving red wine longevity.

New molecularly imprinted polymers for reducing negative volatile phenols in red wine with low impact on wine colour.

4-Ethylphenol (4-EP) and 4-ethylguaiacol (4-EG) formation in red wines by Dekkera/Brettanomyces yeasts reduce significantly wine consumer's acceptability. Polymers with specific adsorption for volatile phenols (VPs) could be a valuable tool for wine producers for removing this negative sensory defect. In this work, a new molecularly imprinted polymer (MIP) was synthesised using ethylene glycol dimethacrylate (EDMA) as cross-linker and ethylene glycol methyl ether acrylate as functional monomers. Although there was observed a competitive binding of the more abundant structurally related phenolic compounds of the wine matrix, it was still able to reduce 38 to 63% the wine VPs, depending on the wine VPs levels, presenting higher performance than the respective non-imprinted polymers (NIP). Sensory analysis of the MIP treated wine resulted in a significant decrease in the phenolic attribute and significant increase of the fruity and floral attributes, with no significant differences in the wine colour perceived by the expert panel. The sensory improvement of the MIP was significantly higher than that observed for the correspondent NIP.

He Impact of Different Tannin to Anthocyanin Ratios and of Oxygen on the Phenolic Polymerisation Over Time in a Wine-like Solution

Colour and phenolic stability during ageing are influenced by the levels of distinct classes of phenolics in young red wines. The ratios between different classes of phenolic compounds also determine the colour and phenolic development of red wines. The present study evaluated the impact of forced oxidation on different anthocyanin/tannin (A/T) extracts and its consequent effect on the colour and phenolic evolution over time. The results showed that higher contents of seed tannins could enhance phenolic polymer formation, especially in the presence of oxygen. The addition of oxygen seemed to favour certain polymerisation reactions between tannins, leading to higher concentrations of monomeric anthocyanins in solution. A slower oxygen consumption was also observed as the phenolic composition of the wine-like extract evolved over time.

HPLC Analysis of Phenols in Negroamaro and Primitivo Red Wines from Salento.

Wine is probably the oldest and still most consumed alcoholic beverage in the world. Nevertheless, it contains several biomolecules with beneficial health effects. Phenols are among them and, in this article, we identified and quantified by HPLC catechin, gallic acid, hydroxytyrosol, quercetin, trans-resveratrol, and syringic acid in Primitivo and Negroamaro red wines from Salento, in Southeast Italy. The concentrations of the analyzed antioxidant molecules were quite high in all varieties. Gallic acid and catechin were the most abundant, but significant concentrations of quercetin, hydroxytyrosol, syringic acid, and trans-resveratrol were also found. Multivariate statistical analysis was also employed to discriminate between Negroamaro and Primitivo wines, suggesting the variables influencing their separation.

Cork powder as a new natural and sustainable fining agent to reduce negative volatile phenols in red wine

In red winemaking, especially those aged in wood barrels, the contamination and growth ofDekkera/Brettanomycesyeasts results in the formation of 4-ethylphenol (4-EP) and 4-ethylguaiacol (4-EG). These volatile phenols (VPs) are responsible for negative aromatic notes like horsy, barnyard, smoky and medicine, decreasing significantly red quality and its commercial value. In this work, cork powder waste was especially prepared and used to remove these negative volatile phenols (4-EP and 4-EG) from spiked red wine. The optimisation of cork powder performance by removal of dichloromethane and ethanol cork extractives, air removal and ethanol impregnation, allowed to obtain 41 to 62% of 4-ethylphenol and 50 to 53% of 4-ethylguaiacol removal from VPs spiked red wine applied at 250 g/hL. There was no significant impact on phenolic acids and monomeric anthocyanins, although being observed a decrease in the headspace aroma abundance (40%). This optimised cork powder allowed to decrease significantly the wine negative phenolic character, bitterness and astringency, recovering the positive fruity and floral sensory attributes. Results show that optimised cork powder can be a good solution for VPs removal, presenting a great potential to be a new oenological fining agent, contributing to the wine industry sustainability.

Spoilage potential of brettanomyces bruxellensis strains isolated from Italian wines

Brettanomyces bruxellensis is an important wine spoilage agent. In this study a population of Brettanomyces strains isolated from Italian wines was thoroughly investigated to evaluate adaptability to wine conditions and spoilage potential. The presumptive isolates of Brettanomyces were identified at species level with 26S rRNA gene sequencing and species-specific PCR, and subsequently subjected to analysis of intra-species variability through the study of intron splice sites (ISS-PCR). Although, some strains were tracked in wines from different regions, extensive genetic biodiversity was observed within the B. bruxellensis population investigated. All strains were evaluated for their growth ability in the presence of ethanol, high sugar content, low pH, different temperatures and sulphur dioxide, using optical density and flow cytometry measurement. The ability of yeasts to produce ethyl phenols in red wines with different chemical compositions was evaluated by means of high performance liquid chromatography with electrochemical detection (HPLC-ECD). The results highlighted wide variability in B. bruxellensis in response to wine limiting factors and in terms of the accumulation of ethyl phenols. As regards this last aspect, the differences found among strains were closely related to chemical composition of wine and strain resistance to environmental stress factors, making a priori evaluation of risk of wine alteration quite difficult. These results suggest that strategies for the control of Brettanomyces should be tailored on the basis of strain distribution and wine characteristics.

Reducing the negative sensory impact of volatile phenols in red wine with different chitosans: Effect of structure on efficiency

“Brett character” is a negative sensory attribute acquired by red wines when contaminating Dekkera/Brettanomyces yeasts produce 4-ethylphenol and 4-ethylguaiacol, known as volatile phenols (VPs), from cinnamic acid precursors. In this study, chitins and chitosans with different structural features, namely deacetylation degree (5–91%) and molecular weight (24–466kDa) were used for the reduction of this sensory defect. Chitins and chitosans decreased 7–26% of the headspace abundance of VPs without changing their amounts in wines. The efficiency of reduction increased with the deacetylation degree and applied dose. Reduction of headspace abundance of VPs by chitosans enabled significant decreases in the negative phenolic and bitterness attributes and increased positive fruity and floral attributes. Results show that chitosan with high deacetylation degrees, including fungal chitosan, which is already approved for use in wines, is an efficient approach for reducing the negative sensory impact of VPs in red wines.

Inhibitory Effects of Red Wine on Lipid Oxidation in Fish Oil Emulsion and Angiogenesis in Zebrafish Embryo.

The capabilities of red wine against lipid oxidation and angiogenesis were evaluated by using a fish oil emulsion system and an in vivo zebrafish embryos model, respectively. The red wine contained 12 different antioxidant phenolics which levels were led by anthocyanins (140.46 mg/L), catechin (55.08 mg/L), and gallic acid (46.76 mg/L). The diversity of the phenolics in red wine was greater than the tea, coffee, or white wine selected as a peer control in this study. The total phenolics concentration of red wine was 305.53 mg/L, although the levels of tea, coffee, and white wine were 85.59, 76.85, and 26.57 mg/L, respectively. The activity of red wine in scavenging DPPH (2,2-diphenyl-1-picrylhydrazyl) free radicals was approximately 4 times higher than the tea and 8 times than the coffee or white wine. The red wine showed the highest capability in preventing long chain PUFA oxidation in the fish oil emulsion. Because of the outstanding antioxidant activity of red wine, the red wine dried extract was used to monitor its inhibitory effect against angiogenesis by using transgenic zebrafish embryos (Tg[fli1:egfp]y1 ) with fluorescent blood vessels. After incubated in 100 μg/mL of the extract solution for 26 h pf, each of the embryos had a lower number of intersegmental vessel than the control embryo. The inhibition rate of red wine extract against growing of angiogenic blood vessel reached 100%.

Solid cation exchange phase to remove interfering anthocyanins in the analysis of other bioactive phenols in red wine.

Bioactive phenols (BPs) are often targets in red wine analysis. However, other compounds interfere in the liquid chromatography methods used for this analysis. Here, purification procedures were tested to eliminate anthocyanin interference during the determination of 19 red-wine BPs. Liquid chromatography, coupled to a diode array detector (HPLC-DAD) and a mass spectrometer (UPLC-MS), was used to compare the direct injection of the samples with solid-phase extractions: reversed-phase (C18) and strong cation-exchange (SCX). The HPLC-DAD method revealed that, out of 13BPs, only six are selectively analyzed with or without C18 treatment, whereas SCX enabled the detection of all BPs. The recovery with SCX was above 86.6% for eight BPs. Moreover, UPLC-MS demonstrated the potential of SCX sample preparation for the determination of 19BPs. The developed procedure may be extended to the analysis of other red wine molecules or to other analytical methods where anthocyanins may interfere.

A simple, cheap and reliable method for control of 4-ethylphenol and 4-ethylguaiacol in red wines. Screening of fining agents for reducing volatile phenols levels in red wines.

Brettanomyces/Dekkera produces 4-ethylphenol (4-EP) and 4-ethylguaiacol (4-EG) from hydroxycinnamic acids that affect the wine aroma and overall quality. A simple, cheap, fast and reliable quantitation method is needed for routine quality control of wines. In this work a simple method based on one simple liquid-liquid extraction with pentane/diethyl ether (2:1) and analysis by GC-MS allow to obtain very good recoveries (98-102%) and low quantification limits (24 and 11μg/L for 4-EP and 4-EG, respectively), well below the sensory threshold for these volatile phenols and with an adequate measurement uncertainty: 70, 1.75 and 78, 1.95 and 1.35μg/L for levels of 1000, 25μg/L for 4-EP and 1000, 25 and 10μg/L for 4-EG, respectively. In addition a screening of eight fining agents (mineral, protein and polysaccharide based) for reducing the levels of these volatile phenols in red wines was performed, and the impact on the physicochemical characteristics of red wines was evaluated. At the levels used, activated carbon was the most efficient fining agent in removing 4-ethylphenol and 4-ethylguaiacol from red wines (57%) resulting in a 75% decrease of headspace concentration of these volatile phenols. Lower reductions were observed when using egg albumin (19%) resulting in a 30% decrease in the headspace concentration. Other fining agents although not reducing the total amount of the volatile phenols present in wine decreased their concentrations in the headspace like isinglass (27%), carboxymethylcellulose (15%) and chitosan (27%). All of these fining agents could be a possibility for treating wine contaminated with 4-ethylphenol and 4-ethylguaiacol.