Eric Wilkes

Effects of Five Metals on the Evolution of Hydrogen Sulfide, Methanethiol, and Dimethyl Sulfide during Anaerobic Storage of Chardonnay and Shiraz Wines

The synergistic effects of Cu, Fe, Mn, Zn, and Al on the evolution of different volatile sulfur compounds (VSCs) in a Chardonnay and a Shiraz wine have been investigated. The evolution of H2S, MeSH, and DMS were influenced by metal addition, and in some instances, a combination of metals was responsible for the largest variation in the concentration of VSCs. The metals and metal combinations associated with significant changes in VSC concentrations in both Chardonnay and Shiraz samples after anaerobic storage were Cu, Fe, Zn, Al, Cu*Fe, Cu*Mn*Al, and Cu*Zn*Al for H2S; Cu, Zn, Fe*Mn, and Cu*Fe*Mn for MeSH; and Al and Zn*Al for DMS. The effect of Cu addition on the evolution of VSCs has previously been shown; however, this investigation has demonstrated that metals other than Cu could also be involved in the catalytic release of VSCs and that the interactions and combinations of metals are important. In some instances, the metal effect was reversed, associated with significant decreases during high oxygen conditions and with significant increases during low oxygen conditions.

The effects of pH and copper on the formation of volatile sulfur compounds in Chardonnay and Shiraz wines post-bottling.

The effects of pH and Cu(2+) treatment on the formation of volatile sulfur compounds (VSCs) were investigated in Chardonnay and Shiraz wine samples. Four VSCs were significantly affected by pH, with lower wine pH associated with decreased hydrogen sulfide (H2S), methanethiol, dimethyl sulfide, and carbon disulfide concentrations. The effects of pH and Cu(2+) on H2S formation from known precursor compounds were subsequently studied in a model wine system. In samples treated with cysteine and glutathione lower pH produced less H2S. Nanoparticle tracking analysis was used to study the effects of variable pH concentrations in a model system containing Cu(2+), tartaric acid, and H2S. Differences in Cu(2)(+)-tartrate complexes particle size and concentration were measured as a function of pH and H2S addition, suggesting the type of complexes formed may affect the binding sites of Cu(2+) available to catalyse the formation of VSCs such as H2S.

The impact of wine components on fractionation of Cu and Fe in model wine systems: Macromolecules, phenolic and sulfur compounds

A variety of techniques have been developed with the ability to measure different forms of metals in wine with the ultimate aim of providing a more accurate indicator of metal induced spoilage of wine. This study was conducted in order to identify which wine components influence the measurement of Cu and Fe in their fractionated and/or electrochemically active forms. The measurement techniques involved detection of labile Cu by stripping potentiometry and fractionation of Cu and Fe by sequential solid phase extraction, with detection by inductively coupled plasma-optical emission spectroscopy. The wine components assessed included those extracted from wine (red wine tannin, white wine protein, white wine polysaccharide, red wine polyphenol, white wine polyphenol), and commercially available monomeric compounds, including phenolic compounds and sulfur-containing compounds. For Cu, only hydrogen sulfide, which is known to induce the formation of Cu(I) sulfide, showed any appreciable influence on the fractionation and electrochemical detection of Cu. This form of Cu was also identified as the major component of red and white wines. For Fe, the fractionation was different for red versus white wine, and influenced significantly by extracted red wine polyphenol, (-)-epicatechin, gallic acid and tartaric acid. The wine components showed more influence on Fe at pH4.00 compared to pH3.25. These results enable a targeted use of these techniques in the assessment of metal-induced spoilage of wine.

Formation of Hydrogen Sulfide in Wine: Interactions between Copper and Sulfur Dioxide.

The combined synergistic effects of copper (Cu2+) and sulfur dioxide (SO2) on the formation of hydrogen sulfide (H2S) in Verdelho and Shiraz wine samples post-bottling was studied over a 12-month period. The combined treatment of Cu2+ and SO2 significantly increased H2S formation in Verdelho wines samples that were not previously treated with either Cu2+ or SO2. The formation of H2S produced through Cu2+ mediated reactions was likely either: (a) directly through the interaction of SO2 with either Cu2+ or H2S; or (b) indirectly through the interaction of SO2 with other wine matrix compounds. To gain better understanding of the mechanisms responsible for the significant increases in H2S concentration in the Verdelho samples, the interaction between Cu2+ and SO2 was studied in a model wine matrix with and without the presence of a representative thiol quenching compound (4-methylbenzoquinone, 4MBQ). In these model studies, the importance of naturally occurring wine compounds and wine additives, such as quinones, SO2, and metal ions, in modulating the formation of H2S post-bottling was demonstrated. When present in equimolar concentrations a 1:1 ratio of H2S- and SO2-catechol adducts were produced. At wine relevant concentrations, however, only SO2-adducts were produced, reinforcing that the competition reactions of sulfur nucleophiles, such as H2S and SO2, with wine matrix compounds play a critical role in modulating final H2S concentrations in wines.

Survey of the Variation in Grape Marc Condensed Tannin Composition and Concentration and Analysis of Key Compositional Factors

Grape marc contains a number of compounds with potential antimethanogenic activity in ruminants, including condensed tannins (CTs). Using direct phloroglucinolysis, a survey of CT chemistry across 66 grape marc samples showed diversity in concentration (6.9 to 138.8 g/kg of dry matter). Concentration was found to be independent of CT composition, although all compositional variables were significantly correlated (P < 0.0001). Twenty samples diverse in CT were selected from a cluster analysis and analyzed for compounds relevant to ruminant digestion and methanogenesis, including metabolizable energy (6.6-12.0 MJ/kg DM), crude protein (3.2-14.4% DM), neutral detergent fiber (18.4-61.4% DM), and ethanol soluble carbohydrates (2.0-40.6% DM). Fatty acid concentrations varied throughout the 20 samples (5.2-184.5 g/kg DM), although fatty acid profile showed two distinct groups. Grape marc varies widely in nutritional value, and in compounds that have been linked with changes in ruminant digestion and methane emissions.

Evaluation of putative precursors of key ‘reductive’ compounds in wines post-bottling

Precursors to hydrogen sulfide (H2S), methanethiol (MeSH), ethanethiol (EtSH), and dimethyl sulfide (DMS) were assessed in wines post-bottling, and the percent yield of VSCs from each precursor determined. Cysteine (Cys) and glutathione (GSH) were associated with small increases in H2S concentrations, with a maximum yield of 0.18% and 1.3%, respectively. Greater yields of H2S were obtained from the combined Cys/GSH and copper treatments in white wine. Copper, acting on unknown precursors, was associated with large increases in H2S in Shiraz wines. Dimethyl disulfide and methyl thioacetate were important precursors to MeSH, and produced maximum yields of 72% and 33%, respectively. Ethyl thioacetate was a key precursor to EtSH, with a maximum yield of 39% obtained. Copper and pH were important in modulating MeSH and EtSH accumulation in wines. A maximum yield of 23% of DMS from S-methylmethionine was obtained, with dimethyl sulfoxide producing significantly less DMS with a maximum yield of only 9.4%.

Analytical strategies for the measurement of different forms of Cu and Fe in wine: Comparison between approaches in relation to wine composition

The speciation of Cu and Fe in wine was assessed by a number of methodologies and those with superior performance were utilised on 49 wines and compared to compositional data. The adopted analytical strategies were stripping potentiometry, HPLC and an extraction (solid-phase or liquid-liquid) followed by atomic absorption or optical emission spectroscopic measurement. Stripping potentiometry was specific for sulfide-bound Cu in wines, and showed that this was the predominant form of Cu. A solid phase extraction technique provided a hydrophobic fraction of Cu that was specific for sulfide-bound Cu in white wine but red wine matrix effects hindered isolation of sulfide-bound Cu. The other methods assessed either perturbed the sulfide-bound Cu or were not sufficiently sensitive. The solid phase extraction method enabled routine fractionation of Fe in red and white wines whilst the additional techniques surveyed either measured total Fe or suffered from background contamination.

Exploiting Compositionally Similar Grape Marc Samples to Achieve Gradients of Condensed Tannin and Fatty Acids for Modulating In Vitro Methanogenesis

Ruminants produce large amounts of the greenhouse gas, methane, which can be reduced by supplementing feed with products that contain anti-methanogenic compounds, such as the solid winemaking by-product, grape marc. The aim of this study was to exploit compositional differences in grape marc to better understand the roles of condensed tannin and fatty acids in altering methanogenesis in a ruminant system. Grape marc samples varying in tannin extractability, tannin size and subunit composition, and fatty acid or tannin concentrations were selected and incubated in rumen fluid using an in vitro batch fermentation approach with a concentrate-based control. Four distinct experiments were designed to investigate the effects on overall fermentation and methane production. Generally, fatty acid concentration in grape marc was associated with decreased total gas volumes and volatile fatty acid concentration, whereas increased condensed tannin concentration tended to decrease methane percentage. Smaller, extractable tannin was more effective at reducing methane production, without decreasing overall gas production. In conclusion, fatty acids and tannin concentration, and tannin structure in grape marc play a significant role in the anti-methanogenic effect of this by-product when studied in vitro. These results should be considered when developing strategies to reduce methane in ruminants by feeding grape marc.