Assessment of the volatile and non-volatile profile of Savatiano PGI wines as affected by various terroirs in Attica, Greece.

Establishing the concept of terroir in wines, combined with the exploitation of native grape varieties, is considered a viable alternative to produce quality wines for increasing interest in the wine market. The aim of this study is the characterization of Protected Geographical Indication (PGI) Savatiano white wines from different regions of Attica (Greece), through the chemical and sensory description of the aroma of wines and the determination of their phenolic profiles. The wines produced with the same vinification protocol were evaluated using the descriptive sensory analysis method while they underwent profiling of volatile and phenolic compounds by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) respectively. The presence of regional differences within the wines was also explored, elaborated with variables that contribute to their differentiation, such as soil type, monthly average temperature, and rainfall. Volatile and sensory profiles of the wines separated the regions and confirmed that sub-regional differences attributed mainly to soil characteristics affect wine aroma. The wines from the east part of Attica were higher in esters, terpenes and higher alcohols with high scores in fruity and blossoms attributes while wines from the north part of Attica presented higher intensity of mineral, nutty and herbaceous attributes. The separations based solely on the phenolic compounds concentration were less clear but a relationship was found between the content of phenolic acids and flavonoids and the studied regions. To our knowledge this is the first characterization of Savatiano PGI wines of Attica generating a fingerprint including chemical composition and sensory aroma characteristics to differentiate wines, combining this pattern with particular sub-regions.

Benzothiadiazole Affects Grape Polyphenol Metabolism and Wine Quality in Two Greek Cultivars: Effects during Ripening Period over Two Years.

Grape berries are one of the most important sources of phenolic compounds, either consumed fresh or as wine. A pioneer practice aiming to enrich grape phenolic content has been developed based on the application of biostimulants such as agrochemicals initially designed to induce resistance against plant pathogens. A field experiment was conducted in two growing seasons (2019-2020) to investigate the effect of benzothiadiazole on polyphenol biosynthesis during grape ripening in Mouhtaro (red-colored) and Savvatiano (white-colored) varieties. Grapevines were treated at the stage of veraison with 0.3 mM and 0.6 mM benzothiadiazole. The phenolic content of grapes, as well as the expression level of genes involved in the phenylpropanoid pathway were evaluated and showed an induction of genes specifically engaged in anthocyanins and stilbenoids biosynthesis. Experimental wines deriving from benzothiadiazole-treated grapes exhibited increased amounts of phenolic compounds in both varietal wines, as well as an enhancement in anthocyanin content of Mouhtaro wines. Taken together, benzothiadiazole can be utilized to induce the biosynthesis of secondary metabolites with oenological interest and to improve the quality characteristics of grapes produced under organic conditions.

Abscisic Acid and Chitosan Modulate Polyphenol Metabolism and Berry Qualities in the Domestic White-Colored Cultivar Savvatiano.

During the last decade, several studies demonstrated the effect of biostimulants on the transcriptional and metabolic profile of grape berries, suggesting their application as a useful viticultural practice to improve grape and wine quality. Herein, we investigated the impact of two biostimulants-abscisic acid (0.04% w/v and 0.08% w/v) and chitosan (0.3% w/v and 0.6% w/v)-on the polyphenol metabolism of the Greek grapevine cultivar, Savvatiano, in order to determine the impact of biostimulants' application in the concentration of phenolic compounds. The applications were performed at the veraison stage and the impact on yield, berry quality traits, metabolome and gene expression was examined at three phenological stages (veraison, middle veraison and harvest) during the 2019 and 2020 vintages. Results showed that anthocyanins increased during veraison after treatment with chitosan and abscisic acid. Additionally, stilbenoids were recorded in higher amount following the chitosan and abscisic acid treatments at harvest. Both of the abscisic acid and chitosan applications induced the expression of genes involved in stilbenoids and anthocyanin biosynthesis and resulted in increased accumulation, regardless of the vintage. Alterations in other phenylpropanoid gene expression profiles and phenolic compound concentrations were observed as well. Nevertheless, they were mostly restricted to the first vintage. Therefore, the application of abscisic acid and chitosan on the Greek cultivar Savvatiano showed promising results to induce stilbenoid metabolism and potentially increase grape defense and quality traits.

Sulfide-bound copper removal from red and white wine using membrane and depth filters: Impacts of oxygen, H2S-to-Cu ratios, diatomaceous earth and wine volume.

Sulfide-bound Cu in wine is a potential contributor to the reductive development of wine. This study examines the effectiveness of filtration for the adsorptive removal of this Cu fraction. The copper concentration in wine before and after filtration was determined by atomic spectroscopy (total) and by stripping potentiometry and/or adsorptive methodologies (Cu fractions). Membrane filters (4.7 cm2) removed significant amounts of sulfide-bound Cu from 10 mL of wine, including 60-80 % removal using nylon membranes, but they could not efficiently remove Cu from larger volumes. Dissolved oxygen concentration in the wine immediately prior to membrane filtration did not impact Cu removal, while a high sulfide-to-Cu(II) ratio did enhance removal. Depth filters incorporating diatomaceous earth with cellulose (45 mm-diameter, 3.5 mm-thickness) showed the most efficient removal of sulfide-bound Cu from wines even after treatment of 3.0 L. The relevance of these laboratory scale filtrations to winery scale filtration is discussed.

The removal of Cu from wine by copolymer PVI/PVP: Impact on Cu fractions and binding agents.

Specific forms of Cu in wine can influence wine flavour and development. The co-polymer polyvinylimidazole/polyvinylpyrrolidone (PVI/PVP) is known to remove Cu from wine, but its impact on different Cu forms is uncertain. In this study, three Cu fractions in white wine were determined by colorimetry and two Cu fractions in red wine were determined by diatomaceous earth depth filtration and atomic spectroscopy. PVI/PVP, with either silica or chitosan formulations, reduced all three fractions of Cu measured in white wines, and sulfide-bound Cu in red wines. The inefficient removal of organic acid-bound Cu in red wines was linked to the higher pH of red wines. After PVI/PVP treatment, wines showed lower concentrations of hydrogen sulfide, but minimal changes in weaker Cu binding agents. These results demonstrate that PVI/PVP efficiently removes the least desirable form of Cu present in wine, along with its detrimental binding agent (i.e., hydrogen sulfide).

Sulfide-binding to Cu(II) in wine: Impact on oxygen consumption rates.

This study investigates the relationship between the two main forms of Cu in wine and their impact on the rate of oxygen consumption. The Cu forms were differentiated by medium exchange constant current stripping potentiometry, which classified the Cu as either bound to sulfide or not. Oxygen consumption rates were determined in red, white and model wines after saturation with oxygen. The results for white wines showed that the oxygen consumption was sensitive to the non-sulfide-bound Cu concentration when ascorbic acid was present, and the first order rates ranged from 0.02 to 0.11 h-1. However, the same was not true for wines without added ascorbic acid that showed little influence of Cu form on oxygen decay rates. Cu forms were also found to significantly change in some wines during the oxygen decay experiment. Ascorbic acid is critical in enabling the form of Cu to significantly influence the oxygen reaction rate in wine.

Changes in Red Wine Composition during Bottle Aging: Impacts of Grape Variety, Vineyard Location, Maturity, and Oxygen Availability during Aging.

This work investigated the influence of grape variety, vineyard location, and grape harvest maturity, combined with different oxygen availability treatments, on red wine composition during bottle aging. Chemometric analysis of wine compositional data (i.e., wine color parameters, SO2, metals, and volatile compounds) demonstrated that the wine samples could be differentiated according to the different viticultural or bottle-aging factors. Grape variety, vineyard location, and grape maturity showed greater influence on wine composition than bottle-aging conditions. For most measured wine compositional variables, the evolution patterns adopted from the viticultural factors were not altered by oxygen availability treatment. However, contrasting evolution patterns for some variables were observed according to specific viticultural factors, with examples including dimethyl sulfide, phenylacetaldehyde, maltol, and ß-damascenone for vineyard locations, 2-methylbutanal, 1,4-cineole, and linalool for grape variety, and methanethiol, methional, and homofuraneol for grape maturity.

Rapid Quantitation of 12 Volatile Aldehyde Compounds in Wine by LC-QQQ-MS: A Combined Measure of Free and Hydrogen-Sulfite-Bound Forms.

This work outlines a rapid novel methodology for the direct quantitation of 12 volatile aldehyde compounds related to oxidative off-flavors in wine, by measuring the combined free and hydrogen-sulfite-bound forms of each aldehyde compound, consisting of four general aldehydes, four Strecker aldehydes, and four (E)-2-alkenals. The methodology requires minimal preparation of wine samples: the addition of internal standards and 6 g/L sulfur dioxide, and filtration prior to quantitation by liquid chromatography-triple quadrupole-mass spectrometery. Overall, the limit of detection, limit of quantification, accuracy (recovery, 97-114%), and precision (repeatability and reproducibility, RSD ≤ 10%) were satisfactory to enable routine measurement of the 12 aldehyde compounds in wine. The methodology was applied to 20 commercial white and red wines from various varieties and vintages. A general trend of higher concentrations of the aldehyde compounds in white wines compared to red wines was observed.

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.

Biological interactions of a calcium silicate based cement (Biodentine™) with Stem Cells from Human Exfoliated Deciduous teeth.

OBJECTIVE: To investigate the biological interactions of a calcium silicate based cement (Biodentine™) with Stem Cells from Human Exfoliated Deciduous teeth (SHED), focusing on viability/proliferation, odontogenic differentiation, biomineralization and elemental release/exchange. METHODS: Biodentine™ specimens were used directly or for eluate preparation at serial dilutions (1:1-1:64). SHED cultures were established from deciduous teeth of healthy children. Viability/proliferation and morphological characteristics were evaluated by live/dead fluorescent staining, MTT assay and Scanning Electron Microscopy. Odontogenic differentiation by qRT-PCR, biomineralization by Alizarin red S staining, while ion elution by Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES). RESULTS: SHED effectively attached within the crystalline surface of Biodentine™ specimens acquiring a spindle-shaped phenotype. Statistically significant stimulation of cell proliferation was induced at day 3 by eluates in dilutions from 1:16 to 1:64. Differential, concentration- and time-dependent expression patterns of odontogenic genes were observed under non-inductive and inductive (osteogenic) conditions, with significant up-regulation of DSPP and Runx2 at higher dilutions and a peak in expression of BMP-2, BGLAP and MSX-2 at 1:8 dilution on day 7. Progressive increase in mineralized tissue formation was observed with increasing dilutions of Biodentine™ eluates. ICP-OES indicated that Biodentine™ absorbed Ca, Mg and P ions from culture medium, while releasing Si and Sr ions from its backbone. SIGNIFICANCE: Biodentine™ interacts through elemental release/uptake with the cellular microenvironment, triggering odontogenic differentiation and biomineralization in a concentration-dependent manner. These results reveal a promising strategy for application of the calcium silicate based cement (Biodentine™) for vital pulp therapies of deciduous teeth in Paediatric Dentistry.

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.

The impact of aging wine in high and low oxygen conditions on the fractionation of Cu and Fe in Chardonnay wine.

Solid-phase extraction has previously been used to fractionate copper and iron into hydrophobic, cationic and residual forms. This study showed the change in fractionated copper and iron in Chardonnay wines with 1-year of bottle aging under variable oxygen and protein concentrations. Wines containing protein in low oxygen conditions induced a decrease (20-50%) in total copper and increased the proportion of the hydrophobic copper fraction, associated with copper(I) sulfide. In contrast, protein stabilised wines showed a lower proportion of the hydrophobic copper fraction after 1-year of aging. In oxidative storage conditions, the total iron decreased by 60% when at high concentration, and the concentration of the residual fraction of both copper and iron increased. The results show that oxidative storage increases the most oxidative catalytic form of the metal, whilst changes during reductive storage depend on the extent of protein stabilisation of the wine.

Improved method for the extraction and chromatographic analysis on a fused-core column of ellagitannins found in oak-aged wine.

A new method for the analysis of ellagitannins observed in oak-aged wine is proposed, exhibiting interesting advantages with regard to previously reported analytical methods. The necessary extraction of ellagitannins from wine was simplified to a single step of solid phase extraction (SPE) using size exclusion chromatography with Sephadex LH-20 without the need for any previous SPE of phenolic compounds using reversed-phase materials. The quantitative recovery of wine ellagitannins requires a combined elution with methanol and ethyl acetate, especially for increasing the recovery of the less polar acutissimins. The chromatographic method was performed using a fused-core C18 column, thereby avoiding the coelution of main ellagitannins, such as vescalagin and roburin E. However, the very polar ellagitannins, namely, the roburins A, B and C, still partially coeluted, and their quantification was assisted by the MS detector. This methodology also enabled the analysis of free gallic and ellagic acids in the same chromatographic run.

Production and Isomeric Distribution of Xanthylium Cation Pigments and Their Precursors in Wine-like Conditions: Impact of Cu(II), Fe(II), Fe(III), Mn(II), Zn(II), and Al(III).

This study establishes the influence of Cu(II), Fe(II), Fe(III), Zn(II), Al(III), and Mn(II) on the oxidative production of xanthylium cations from (+)-catechin and either tartaric acid or glyoxylic acid in model wine systems. The reaction was studied at 25 °C using UHPLC and LC-HRMS for the analysis of phenolic products and their isomeric distribution. In addition to the expected products, a colorless product, tentatively assigned as a lactone, was detected for the first time. The results show the importance of Fe ions and a synergistic influence of Mn(II) in degrading tartaric acid to glyoxylic acid, whereas the other metal ions had minimal activity in this mechanistic step. Fe(II) and Fe(III) were shown to mediate the (+)-catechin-glyoxylic acid addition reaction, a role previously attributed to only Cu(II). Importantly, the study demonstrates that C-8 addition products of (+)-catechin are promoted by Cu(II), whereas C-6 addition products are promoted by Fe ions.

The effect of supplementation with three commercial inactive dry yeasts on the colour, phenolic compounds, polysaccharides and astringency of a model wine solution and red wine.

BACKGROUND: Nowadays supplementing red wines with commercial inactive dry yeasts is a widespread practice in winemaking because it leads to better balanced wines through increased mouthfeel and smooth astringency. The aim of this article is to study, in a red wine and in a model wine solution, how supplementation with three commercial inactive dry yeasts affects chemical composition and astringency. This will give us a better understanding of the action mechanism involved. RESULTS: The results suggest that this action mechanism is related to two different phenomena. The first is that inactive yeasts release polysaccharides and oligosaccharides which can increase mouthfeel and inhibit interactions between salivary protein and tannins. The second is that they have a direct effect on the precipitation or absorption of proanthocyanidins, especially the larger polymers, which have been described as the most astringent. CONCLUSION: It can be concluded that supplementation with inactive yeasts is indeed a useful tool for smoothing the astringency of red wines. © 2016 Society of Chemical Industry.

Measurement of labile copper in wine by medium exchange stripping potentiometry utilising screen printed carbon electrodes.

The presence of copper in wine is known to impact the reductive, oxidative and colloidal stability of wine, and techniques enabling measurement of different forms of copper in wine are of particular interest in understanding these spoilage processes. Electrochemical stripping techniques developed to date require significant pretreatment of wine, potentially disturbing the copper binding equilibria. A thin mercury film on a screen printed carbon electrode was utilised in a flow system for the direct analysis of labile copper in red and white wine by constant current stripping potentiometry with medium exchange. Under the optimised conditions, including an enrichment time of 500s and constant current of 1.0µA, the response range was linear from 0.015 to 0.200mg/L. The analysis of 52 red and white wines showed that this technique generally provided lower labile copper concentrations than reported for batch measurement by related techniques. Studies in a model system and in finished wines showed that the copper sulfide was not measured as labile copper, and that loss of hydrogen sulfide via volatilisation induced an increase in labile copper within the model wine system.

Impact of wine production on the fractionation of copper and iron in Chardonnay wine: Implications for oxygen consumption.

Copper and iron in wine can influence oxidative, reductive and colloidal stability. The current study utilises a solid phase extraction technique to fractionate these metals into hydrophobic, cationic and residual forms, with quantification by ICP-OES. The impact of aspects of wine production on the metal fractions was examined, along with the relationship between metal fractions and oxygen decay rates. Addition of copper and iron to juice, followed by fermentation, favoured an increase in all of their respective metal fractions in the wine, with the largest increase observed for the cationic form of iron. Bentonite fining of the protein-containing wines led to a significant reduction in the cationic fraction of copper and an increase in the cationic form of iron. Total copper correlated more closely with oxygen consumption in the wine compared to total iron, and the residual and cationic forms of copper provided the largest contribution to this impact.

Oxygen consumption by oak chips in a model wine solution; Influence of the botanical origin, toast level and ellagitannin content.

The botanical origin, toast level and ellagitannin content of oak chips in a model wine solution have been studied in terms of their influence on oxygen consumption. French oak chips released significantly higher amounts of ellagitannins than American oak chips at any toast level. The release of ellagitannins by oak chips decreased as the toast level increased in the French oak but this trend was not so clear in American oak. Oxygen consumption rate was clearly related to the level of released ellagitannins. Therefore, oak chips should be chosen for their potential to release ellagitannins release should be considered, not only because they can have a direct impact on the flavor and body of the wine, but also because they can protect against oxidation.

Influence of the botanical origin and toasting level on the ellagitannin content of wines aged in new and used oak barrels.

The influence of the botanical origin (French oak: Quercus petraea and American oak: Quercus alba) and different toasting levels (light, medium and heavy) on the ellagitannin content of wines aged in oak barrels has been studied. This took place in two consecutive vintages in order to study what happens after the barrels have been already used for one year with another wine. This study was carried out with two red wines (Cabernet Sauvignon) and with two decolored white wines (Macabeo) from vintages 2012 and 2013 in order to work with a simpler matrix which facilitates chemical analysis. The results show that the botanical origin, toasting level and the number of times that the barrels have been used exert a major influence on the final ellagitannin concentration. In general, the behavior of all the individual ellagitannins was very similar to that described for the total ellagitannins. Briefly, the levels of total ellagitannins concentration in the decolored white wine aged for 12months in new French barrels ranged between 31.2mg/L in the lightly toasted and 4.7mg/L in the heavy toasted. In contrast, these levels were quite lower in American new barrels ranging between 3.6mg/L and 0.9mg/L. Finally, the total ellagitannin concentration decreased an average of 63% in the wines aged in the one year used barrels.

Influence of grape maturity on the foaming properties of base wines and sparkling wines (Cava).

BACKGROUND: The aim of this paper is to determine the main factors that influence the foaming properties of base wines and sparkling wines and, in particular, the role played by the maturity of the grapes in the controlled designation of origin (CDO) Cava. Macabeo, Xarel.lo, Parellada and Chardonnay grapes were harvested at two different maturity levels in two vintages. The first harvest was selected because of its desirable titratable acidity and the second because of its optimal sugar concentration. The mono-varietal base wines and standard blends were transformed into sparkling wines and aged for 14 months. RESULTS: The results indicate that the main factors affecting foam properties are ethanol content, high molecular weight polysaccharides and gluconic acid (which have a negative effect), and proteins (which have a positive effect). CONCLUSION: The main conclusion of this study is that the foaming properties of wines can be improved if harvest dates are advanced.