Effects of pulsed electric fields on the extraction of phenolic compounds during the fermentation of must of Tempranillo grapes

The effects of a pre-treatment of the grape skin with electric field treatments (PEF) at 5 and 10 kV/cm on the evolution of colour, anthocyanin and index of total polyphenols during the Tempranillo grape vinification and on the wine characteristics after fermentation have been investigated.Results showed that the permeabilization of the grape skin by application of a PEF treatment at room temperature caused an increment of the colour intensity, anthocyanin content and of total polyphenolic index with respect to the control during all the vinification process. Generally along the maceration process the values of these three parameters increased when the electric field strength raised from 5 to 10 kV/cm.Colour intensity and anthocyanin content of the wine significantly increased when the PEF treatment was applied at 10 kV/cm. However, the total phenolic index increased considerably with the application of a PEF treatment at 5 kV/cm. A further increment on the electric field strength did not appreciably augment this attribute. PEF did not affect the ratio between the components of the red wine colour (tint and yellow, red and blue components) and other wine characteristics such as alcohol content, total acidity, pH, reducing sugar concentration and volatile acidity.These results indicate that the PEF treatment previous to the maceration step in the winemaking process of the red wine can contribute to reduce the duration of the maceration during vinification or to increase the quantity of these attributes in the final wine.

Industrial relevance

This study investigates the effect of the application of a PEF pre-treatment of the grape skin on the evolution of colour, anthocyanin and index of total polyphenols during the Tempranillo grape vinification. Results obtained indicate that the application of a PEF treatment preceding the maceration step in the winemaking process of red wine could contribute to reduce the duration of the maceration during vinification or to increase the quantity of these parameters in the final wine. Latter effect could be of interest in grapes poor in polyphenols.     

Effect of the prefermentative addition of copigments on the polyphenolic composition of Tempranillo wines after malolactic fermentation

The influence of the prefermentative addition of copigments and different winemaking technologies on the polyphenolic composition of Tempranillo red wines after malolactic fermentation was studied. Six experiments dealing with the prefermentative addition of caffeic acid, rutin, (+) catechin, white grape skin tannin, white grape seed tannin and control wines were realised. Three different winemaking technologies (traditional vinification, prefermentative cold maceration at 6–8 °C and cold soak at 0–2 °C with dry ice) were studied. Prefermentative addition of copigments increases anthocyanin copigmentation reactions and produces wines with a greater colour, a higher anthocyanin concentration, a superior contribution of anthocyanins to the colour of the wine, a superior percentage of tannins polymerised with polysaccharides and less astringency. Cold prefermentative maceration increases the extraction of polyphenols, the anthocyanin copigmentation reactions and the polymerisation reactions between tannins and polysaccharides. The effectiveness of the combination of copigments and prefermentative maceration treatments was demonstrated by the increase of the concentration of the polyphenolic compounds.     

Antioxidant activity in relation to the phenolic profile during the winemaking of sweet wines Vitis vinifera cv. Cabernet Sauvignon

The phenolic profile during the winemaking of Cabernet Sauvignon sweet wines has been studied, including the stages of controlled grape drying and the posterior maceration of the fortified musts with their skins. Each phenolic family has been correlated individually with the antioxidant activity measured by the 2,2,‐diphenylpicrylhydrazyl assay. Linear regressions with a confidence level superior to 99% have been found with hydroxybenzoic acids, esters of hydroxycinnamic acids, flavan‐3‐ol derivatives, tannins, anthocyanin adducts, polymers and polymeric pigments. With a multiple correlation, the antioxidant properties of musts and sweet wines were explained jointly by the families of polymeric pigments and flavan‐3‐ol derivatives. The musts during the drying process and the wines during the maceration presented a different phenolic composition and were distinguished using a cluster analysis and a principal component analysis.     

In vitro and in vivo antioxidant activity of musts and skin extracts from off-vine dried Vitis vinifera cv. “Tempranillo” grapes

The phenolic composition and antioxidant activity of musts and skin extracts from Tempranillo grapes dried for variable lengths of times were examined here. Both were found to increase in the grape must with increasing drying time, but the opposite trend was observed in the skin extracts. The antioxidant activity of the must and skin was largely due to anthocyanins. Also, the anthocyanins fraction exhibited the highest antioxidant activity per gram of phenolic compound in the musts and skin extracts at the end of the drying process, polymeric pigments and high-molecular-weight procyanidins were also prominent in this respect. In vivo antioxidant activity in terms of protein oxidation and survival of Saccharomyces cerevisiae cells previously incubated with 10 μg/mL of polyphenols from the musts and skins and exposed to hydrogen peroxide was also measured. Both extracts exerted a protective effect against oxidative stress, the effect increasing with increasing dehydration time and being somewhat higher for skin than for must. Thus, must and skin from dehydrated grapes possess antioxidant properties, which allow the latter, a by-product of sweet wine, to be used as a natural source of antioxidants.     

Evolution of polyphenolic compounds in red wine from Cabernet Sauvignon grapes processed by pulsed electric fields during aging in bottle

The evolution of the Folin–Ciocalteu index, colour intensity and the principal individual polyphenols throughout 12 months of aging in bottles of red wine from Cabernet Sauvignon grapes treated and untreated by pulsed electric fields (PEF) have been compared. PEF technology allowed bottling of wines with higher Folin–Ciocalteu index, colour intensity and individual polyphenols concentrations using a shorter maceration time. For both wines, aging did not affect the colour intensity, whereas the Folin–Ciocalteu index decreased. The evolution of the principal individual polyphenols during the 12 months of storage followed a similar pattern in both wines. At the end of aging, there were no significant differences observed in the content of monomeric anthocyanins between both wines, however, the content of flavan-3-ols, flavonols and hydroxycinnamic acids and derivatives was higher in PEF treated wine. These results indicate that PEF could be a suitable technology for obtaining wines with higher phenolic content or reducing the maceration time during vinification.     

The influence of ripeness grade on the composition of musts and wines from Vitis vinifera cv. Tempranillo grown in a warm climate

Fruit ripeness has an influence on the phenolic and aromatic composition of grapes and musts and, therefore, on the sensorial characteristics of the resulting wine. The aim of the work described here was to assess the influence of the harvest date on the analytical composition of the musts and wines from the Vitis vinifera cv. Tempranillo grape grown in a warm climate. A hydroalcoholic medium was employed to extract the musts and simulate a traditional maceration process. Phenolic compounds were analysed by colorimetric methods and aroma compounds by gas chromatography/mass-spectrometry. Furthermore, the wines were tested by a sensory panel. A good correlation was obtained between the parameters measured in the must and in the corresponding wine, as well as in the scores recorded in the sensory analysis. In summary, grape ripening was found to have a marked influence on the wine composition and it was possible to establish the key parameters that need to be measured in the must during ripening to identify the optimal harvest date to obtain the best quality wine.     

Changes in chromatic characteristics and phenolic composition during winemaking and shelf-life of two types of red sweet sparkling wines

The Brachetto variety is the most important aromatic red grape used for the production of sweet sparkling wines in Italy. The phenolic composition and chromatic characteristics were studied in base and red sweet sparkling wines made from this variety. The present work represents the first study on sparkling wines manufactured with Brachetto grapes. The amount of pigments extracted in the base wine was low as a consequence of the necessity for short maceration time (48 h) and low alcohol content (< 3.5% v/v). The second fermentation (prise de mousse) caused a pigment content decrease, accompanied by both a color intensity decrease and a tonality increase. In terms of phenolic compounds and chromatic characteristics, lightly sparkling wines (final bottle pressure < 1.7 bar) agreed with fully sparkling wines (final bottle pressure > 3.0 bar) at the end of the second fermentation and, therefore, the chromatic quality was independent on the winemaking methodology used, excepting for lightness and color intensity.Response surface methodology was applied to predict the effect of two independent variables, namely time and temperature of storing, on the phenolic composition and chromatic properties in both lightly and fully sparkling wines. So, it is possible to evaluate the development of two types of sweet sparkling wines during their ageing in bottle and their commercial shelf-life. A central composite design (CCD) and response surface methodology (RSM) were used for this purpose. Quantitative changes were observed in the color parameters. Although the time variable strongly influenced them, the two sparkling wines were stable enough only at low temperatures after 12 months of ageing. During the wine ageing in bottle, an increase in the formation of polymeric red pigments can be suggested taking into account the losses observed in monomeric anthocyanins. The results obtained showed that the color of fully sparkling wines was more stable than that of lightly ones and, therefore, the shelf-life of the former ones could be longer.     

Influence of winemaking techniques on the resveratrol content, total phenolic content and antioxidant potential of red wines

In this paper, the influence of winemaking techniques and cultivars on the resveratrol content, total phenolic content and antioxidant potential of red wines was studied. Wines were made from the cultivars Merlot, Cabernet Sauvignon, Pinot Noir and Prokupac. Applied winemaking technologies included thermovinification and separation of must from pomace. The analysis of trans- and cis-resveratrol in wines was performed by HPLC, while the total phenolic content and antioxidant potential was determined by spectrophotometric methods. The total resveratrol content in analysed samples ranged from 0.35 to 4.85 mg/l; Merlot wines had the highest average resveratrol content, while the lowest was found for native cultivar Prokupac. Although the resveratrol content depended on grape variety, correlation between the winemaking technology applied and the resveratrol level in wines was not observed. The total phenolic content (TPC) varied from 544.4 to 1410.4 mg/l expressed as gallic acid equivalents, and the antioxidant potential, assayed by DPPH in terms of SC₅₀ (mean scavenging concentration), ranged from 0.58 to 2.91 μl/ml. Obtained results showed that thermovinificated wine samples had higher amount of phenolic compounds. Significant negative correlation was observed (p < 0.05) between the total phenolic content and SC₅₀, but there was no significant correlation (p > 0.05) between the resveratrol content and SC₅₀ or TPC. This study could contribute to the establishment of optimal conditions for producing red wines which contain more beneficial phenolic compounds.     

Application of high‐power ultrasounds during red wine vinification

Wine colour is one of the main organoleptic characteristics influencing its quality. It is of special interest in red vinifications due to the economic resources that wineries have to invest for the extraction of the phenolic compounds responsible for wine colour, compounds that are mainly located inside the skin cell vacuoles, where the volatile compounds are also found. The transfer of phenolic compounds from grapes to must during vinification is closely related to the type of grapes and the winemaking technique. During traditional winemaking, grapes are crushed and skin macerated for several days, with pumps overs to facilitate the colour extraction. To increase this extraction, some chemical (maceration enzymes) or physical technologies (thermovinification, cryomaceration, flash‐expansion) can be applied. In this work, a new methodology has been tested. This methodology consists in the application of high‐power ultrasounds to crushed grapes to increase the extraction of phenolic compounds. Crushed grapes were treated with this non‐thermal technology and vinified, with 3, 6 and 8 days of skin maceration time, and the results were compared with a control vinification, where crushed grapes were not subjected to any treatment and were skin macerated during 8 days. The wine chromatic characteristics (determined spectrophotometrically) and the individual phenolic compounds (anthocyanins and tannins, determined by HPLC) were followed during the maceration period, at the end of alcoholic fermentation and after two months in bottle. Also, the wine volatile compounds were determined by GC‐MS. The wines made with ultrasound‐treated grapes showed differences with the control wine, especially regarding total phenol content and tannin content. The wines elaborated with sonicated grapes and with only three days of skin maceration time presented similar concentration of anthocyanins and twice the concentration of tannins than control wines elaborated with 8 days of skin maceration.     

Influence of Pulsed Electric Fields processing at high-intensity electric field strength on the relationship between anthocyanins composition and colour intensity of Merlot (Vitis vinifera L.) musts during cold maceration

Merlot grapes have high proportion of anthocyanins protected inside cell vacuoles but the thick skins limit their extractability. This study investigated the effect of a continuous Pulsed Electric Fields (PEF at 500 kg/h) system operated at high-intensity electric field strengths (>30 kV/cm) combined with energy inputs ranging from 4.7 to 49.4 kJ/L on anthocyanins extraction and colour intensity of Merlot musts during a 4-day cold maceration. Considerable amount of anthocyanins with different chemical features was extracted instantaneously due to PEF treatment at high-intensity electric field strength, in which the amount was higher than those extracted from non PEF-treated Merlot followed by 4 days maceration. During maceration, malvidin derivatives were the most significant anthocyanins contributing (r > 0.71, p < 0.05) to the changes in colour intensity of PEF-treated musts. This finding suggests that malvidin derivatives are good indicators to predict the efficiency of specific PEF process condition in reducing the length of cold maceration while achieving maximum colour intensity in the musts. This research is important either for the wine and beverage industries.Industrial relevance textPrevious PEF studies mainly investigated the effect of low-intensity electric field strength (<7 kV/cm) on various grapevine varieties, the current study investigated the effect of PEF on grapes at high-intensity electric field strength (>30 kV/cm) using Marx generator for the first time in the literature. Marx generator has a tendency to produce high-intensity electric field strength, up to 80 kV/cm, but the impact on plant materials intended for food application are not well studied. In this study, Merlot grapes were continuously treated with PEF at high-intensity electric field strength at a flow rate of 500 kg/h while many previous studies conducted trial at approximately 100–200 kg/h in conjunction with low-intensity electric field strength (<7 kV/cm). Therefore, it seems necessary to advance the current application of PEF for grape processing to high-intensity electric field strength in order to cope with or fit into the standard production rate (>200 kg/h) at the commercial scale. In this study, we have successfully showed that high-intensity electric field strengths (33.1 and 41.5 kV/cm) of PEF were efficient (i.e. instantaneous extraction) in extracting all types of anthocyanins with different chemical features from the thick-grape skins of Merlot. Therefore, grape juice with more elaborated anthocyanin composition and intense colour was produced at a large scale immediately after PEF. The extracted anthocyanins were found to be stable over time after PEF treatment in which the issues on anthocyanin stability and colour development have never been considered in many previous studies, but are important quality aspects that beverage industry desired.     

Stilbene levels and antioxidant activity of Vranec and Merlot wines from Macedonia: Effect of variety and enological practices

The content of the stilbenes trans-resveratrol and piceid as well as the antioxidant activity of Macedonian red wines from the two main grape varieties Vranec and Merlot have been evaluated. Тhe effects of time of maceration, type of yeast and the level of sulphur dioxide applied on stilbene content and antioxidant activity have been studied. The most important factor in winemaking technology is the maceration time since the highest concentrations of trans-resveratrol, piceid and highest antioxidant activity were found following 6 and 10 days of maceration. Concerning the yeast type, higher concentrations of trans-resveratrol and piceid have been obtained with French yeast “Levuline CHP” in comparison to Macedonian yeast “Vinalco”. In contrast, the higher antioxidant activity of wines from both varieties of grapes was observed by application of Macedonian yeast “Vinalco”.     

Effect of skin contact on bound aroma and free volatiles of Listán blanco wine

Listán blanco is the major grape variety of the Canary Islands. Aroma precursors in musts and free volatiles in wines and their variations due to the skin contact process (10 °C and 0–12 h of maceration) were determined in samples of the Listán blanco grape variety. Aroma precursors of must were isolated by absorption on XAD-2 resin and the aglycone liberated by means of acid hydrolysis. Free volatiles from wines and from aroma precursors were extracted with dichloromethane and then analysed by GC-MS. The results of two consecutive vintages show that only aroma precursors of some terpenes and phenols may affect the final wine aroma. However, the majority of the free compounds of the wines did not present differences in content that could be related to the skin contact process.     

Anthocyanin profile of young red wines of Tannat,Syrah and Merlot made using maceration enzymes and cold soak

Anthocyanins are the main pigments of young red wines. The aim of this work was to evaluate the anthocyanin composition of red wines produced by alternative procedures of winemaking and different grape varieties. For this purpose, wines of Tannat, Syrah and Merlot were made by traditional maceration (TM), addition of maceration enzymes (ENZ) and cold soak (CS). Syrah wines had the highest proportions of malvidin (86.0 ± 1.0) and peonidin (5.5 ± 0.4), while Tannat wines had the highest of delphinidin (6.9 ± 0.8), petunidin (13.2 ± 0.6) and nonacylated glucosides (73.6 ± 0.6). Cold soak increased the anthocyanins contents in Tannat and Merlot wines. In turn, maceration enzymes affect the anthocyanin profile and the stability of the colour of wines. Discriminant analysis of data showed significant separation of the wines of each variety as the anthocyanin profile showed the varietal fingerprint, despite the different winemaking procedures.     

Improvement of winemaking process using pulsed electric fields at pilot-plant scale. Evolution of chromatic parameters and phenolic content of Cabernet Sauvignon red wines

The influence of the application of a continuous pulsed electric fields treatment (PEF treatment) to grape pomace on the evolution of colour and phenolic content of Cabernet Sauvignon red wines has been investigated. Wine from grape treated by PEF presented at the end of alcoholic fermentation higher colour intensity (CI), total polyphenol index (TPI) and total anthocyanic content (TAC) than control wine. This effect was observed even although maceration time for PEF wine was 48 h shorter than for control. Differences remained during malolactic fermentation and maturation. After 4 months of aging in bottle, CI, TPI and TAC of PEF wine were 38%, 22% and 11% higher respectively than the control. HPLC phenolic profiles of wines were qualitatively similar, without detecting a selective effect on any phenol. No significant differences in sensory attributes between wines were detected. Results indicate that PEF is a promising technology in red winemaking for reducing the maceration time and increasing colour and phenolic extraction.     

Determination of flavan-3-ols and trans-resveratrol in grapes and wine using HPLC with fluorescence detection

Concentrations of trans-resveratrol, catechin and epicatechin were analyzed in musts and wines produced from seven red and four white grape cultivars from various wine growing regions of Turkey. Phenolics were quantified using an HPLC method optimized for the separation of wine phenolics. Wine samples contained higher phenolics levels than the corresponding musts. With the exception of Semillion, white wines and musts contained lower concentrations of phenolics than red wines and musts. However, the white cultivar Semillion had the highest concentrations of catechin and epicatechin among all wine and must samples. Semillion wine catechin and epicatechin were 13.7 and 11.8 mg/L, respectively. The highest level of trans-resveratrol among the white cultivars was found in Narince wine (1.93 mg/L). Within the red wine and must cultivars, Bo?azkere, Öküzgozü, and Cabernet contained the highest concentrations of flavan-3-ols and trans-resveratrol. Catechin was the major phenolic in all wines and most musts. Epicatechin was the major phenolic in 6 of the 11 must samples, but none of the wine samples. trans-Resveratrol was generally found in lowest concentrations in both wines and musts.     

Polyphenols and colour variability of red wines made from grapes harvested at different ripeness grade

Several red wines were elaborated in order to study the influence of the grape harvesting date (degree of maturity of the grape) on their chromatic characteristics and polyphenolic contents. Wines made from two grape varieties, at three different harvesting dates and from two consecutive vintages, were selected for this study. The results showed that the harvesting date of grapes (directly correlated with the degree of maturity of the grapes), influenced the chromatic characteristics of the wines, although their polyphenolic compositions were clearly different, especially in Cabernet Sauvignon wines. In general, higher intensities of blue or violet tones were detected in wines made from the grapes collected on the second harvesting date, in which the ratios anthocyanins/proanthocyanidins and anthocyanins/(proanthocyanidins + catechins) were the lowest. These ratios are proposed as probable indicators of the aptitude for wine ageing.     

Influence of the heterogeneity of grape phenolic maturity on wine composition and quality

Nowadays, consumers demand red wines with deep colour, soft tannins and fruit scents, but these wines can only be obtained from grapes with complete phenolic maturity. Diverse methods have been proposed for measuring phenolic maturity. However, all these methods only provide the average value and do not consider any possible heterogeneity. Throughout ripening, grapes were separated according to their density, which revealed the existence of a large heterogeneity. Grapes at harvest were also separated by density in three groups. The higher the density of the grapes the higher ethanol content, pH, colour intensity, total phenolic index and anthocyanin and proanthocyanidin concentrations, and the lower the titratable acidity and bitterness of the wines. When the grapes were denser the wines were also better balanced in flavour and mouthfeel sensation. These results suggest that grape heterogeneity may influence the final wine composition and quality and therefore it should be considered at harvest.     

Pre-bottling use of dehydrated waste grape skins to improve colour,phenolic and aroma composition of red wines

Different dehydrated waste grape skins from the juice industry were added into aged and young red wines as an innovative way of compensating for colour loss before bottling. After addition of grape skins, colour intensity of wines increased a mean 11% and a maximum of 31% with predominance of the red component. Total polyphenols mean increase was 10% with a maximum value of 20%. Analysis of low molecular weight phenolic compounds by HPLC–DAD showed a significant (p < 0.05) content increase of the bioactive compounds gallic acid, (+)-catechin, (−)-epicatechin, and (E)-resveratrol. Anthocyanins content also increased at an average of 50 mg/l. The volatile profile of wines analysed by SBSE–GC–MS was only moderately influenced by the treatments. Mixtures of dehydrated waste grape skins were useful to improve the colour and polyphenol profile of red wines, considering them a useful tool for correcting colour loss before bottling.