Microwave-assisted maceration and stems addition in Bonarda grapes: Effects on wine chemical composition over two vintages.

Bonarda, the second red grape variety in Argentina, produces high yields per hectare generating, in several cases, low quality wines. Microwave-assisted extraction (MW) is a novel extraction technique for winemaking, widely applied in other foods. Stems addition (S) during vinification can be a sustainable technology for phenolic and aroma contribution without additional cost. Therefore, this study aimed to evaluate the combined effect of MW application with stem additions in different conditions, before fermentation, on the chemical composition of Bonarda wines. During two consecutive vintages (2018 and 2019), the experimental design consisted of ten treatments (two factors) by triplicate. Two maceration strategies (Factor 1) were applied [control (C), and microwaved-assisted extraction after grape crushing (MW; 2450 MHz, 7600 W, 45-50 °C)], combined with five stem-contact conditions (Factor 2) [control without stems (WS), 50% stems addition (S50), 50% stems addition + MW of the stems (S50MW; 2450 MHz, 7600 W, 60 °C), 100% stems addition (S100), 100% stems addition + MW (S100MW)]. Wines were analyzed for basic chemistry, phenolic composition and color parameters, polysaccharides, and aroma profiles. The 2018 wines showed higher pH and lower volatile acidity with stem additions and MW application in both matrices (grapes and stems). Stem additions increased tannin content by >55% (S100) and by >25% for the other treatments; while MW, mostly in the 2018 season, consistently improved phenolic extraction and polymeric pigments formation, enhancing wine color (greater saturation) and intensifying violet hue. The behavior observed in 2019 was similar, with a more marked effect of MW on wine color (C*ab and polymeric pigments). Likewise, combined strategies increased polysaccharides extraction and modified the volatile profile of wines. The reported results are promising and are considered the first advance in the knowledge of the impact of the proposed combined strategies on the chemical composition of red wines.

Mutagenesis, screening and isolation of Brettanomyces bruxellensis mutants with reduced 4-ethylphenol production.

The use of non-conventional yeast species to obtain interesting flavors and aromas has become a new trend in the fermented beverages industry. Among such species, Brettanomyces bruxellensis (B. bruxellensis) has been reported as capable of producing desirable or at least singular aromas in fermented beverages like beer and wine. However, this yeast can also produce an aromatic defect by producing high concentrations of phenolic compounds like, 4-ethylguaiacol and particularly 4-ethylphenol (4-EP). In the present study, we designed a mutant screening method to isolate B. bruxellensis mutants with reduced 4-EP production. More than 1000 mutants were screened with our olfactory screening method, and after further sensory and chemical analysis we were able to select a B. bruxellensis mutant strain with a significant reduction of 4-EP production (more than threefold) and less phenolic perception. Notably, the selected strain also showed higher diversity and concentration of ethyl esters, the most important group of odor active compounds produced by yeasts. Based on these results, we consider that our selected mutant strain is a good candidate to be tested as a non-conventional yeast starter (pure or in co-inoculation) to obtain wines and beers with novel aromatic properties.

Impact of closure type and storage temperature on chemical and sensory composition of Malbec wines (Mendoza, Argentina) during aging in bottle.

Malbec is the flagship variety of Argentina mainly due to its high oenological value and plasticity to obtain different wine styles. During bottled aging, the chemical and organoleptic composition of wines is subject to changes depending on the aging conditions (closure, oxygen level, temperature, time). However, the combined effect of these factors on chemical composition and organoleptic characteristics of Malbec wines has not been studied yet. Wines were bottled with screw cap and natural cork and were kept in chambers at 15 °C and 25 °C for 2 years. Sampling was performed at 2, 4, 6, 9, 12, 15, 18, 21 and 24 months. Concentrations of free sulfur dioxide, dissolved oxygen, anthocyanins, tannins, esters, volatile phenols, organic acids, and color saturation decreased during the storage process. While, the formation of polymeric pigments, the color attributes (lightness and hue) and the levels of alcohols, norisoprenoids, furanoids and terpenoids increased. At 24 months, Malbec wines were organoleptically different. Wines kept at 15 °C were associated with high sensory perceptions in color intensity and violet tint, those presented a positive correlation with free sulfur dioxide, tannins, and anthocyanins levels. On the contrary, wines aged at 25 °C were linked with high sensory perceptions of dried vegetative and dried fruit aromas. These descriptors were positively correlated with norisoprenoids, furanoids, and terpenoids. In general, the chemical composition and organoleptic attributes of bottled Malbec wines (Mendoza, Argentina) were stable respect closure type employed, but highly sensitive to the combined effect of time and storage temperature. This finding is key to making decisions about the wine style searched, and costs (e.g. refrigeration) involved in the conservation period until consumption.

Determination of alkaloids in onion nectar by micellar electrokinetic chromatography.

Nectar is the most important floral reward offered by plants to insects. Minor components such as alkaloid compounds in nectar affect bee foraging, with great influence in seed production. CE is an advantageous tool for the analysis of unexplored samples such as onion nectar due to the limited amounts of samples. Considering the importance of these compounds, a simultaneous determination of nicotine, theophylline, theobromine, caffeine, harmaline, piperine in onion nectar by MEKC-UV is herein reported. The extraction of alkaloid compounds in nectar was performed by SPE using a homemade miniaturized column (C18 ). Effects of several important factors affecting extraction efficiency as well as electrophoretic performance were investigated to acquire optimum conditions. Under the proposed conditions, the analytes can be separated within 15 min in a 50 cm effective length capillary (75 µm id) at a separation voltage of 20 kV in 20 mmol/L sodium tretraborate, 100 mmol/L SDS. The amount of sample requirement was reduced up to 2000 times, when compared to traditional methods, reaching limits of detection as low as 0.0153 ng/L. For the first time, this study demonstrates that there are marked qualitative and quantitative differences in nectar alkaloids between open pollinated and male sterile lines (MSLs) and also within MSLs.

Enzymatic activities produced by mixed Saccharomyces and non-Saccharomyces cultures: relationship with wine volatile composition.

During certain wine fermentation processes, yeasts, and mainly non-Saccharomyces strains, produce and secrete enzymes such as ß-glucosidases, proteases, pectinases, xylanases and amylases. The effects of enzyme activity on the aromatic quality of wines during grape juice fermentation, using different co-inoculation strategies of non-Saccharomyces and Saccharomyces cerevisiae yeasts, were assessed in the current study. Three strains with appropriate enological performance and high enzymatic activities, BSc562 (S. cerevisiae), BDv566 (Debaryomyces vanrijiae) and BCs403 (Candida sake), were assayed in pure and mixed Saccharomyces/non-Saccharomyces cultures. ß-Glucosidase, pectinase, protease, xylanase and amylase activities were quantified during fermentations. The aromatic profile of pure and mixed cultures was determined at the end of each fermentation. In mixed cultures, non-Saccharomyces species were detected until day 4-5 of the fermentation process, and highest populations were observed in MSD2 (10% S. cerevisiae/90% D. vanrijiae) and MSC1 (1% S. cerevisiae/99% C. sake). According to correlation and multivariate analysis, MSD2 presented the highest concentrations of terpenes and higher alcohols which were associated with pectinase, amylase and xylanase activities. On the other hand, MSC1 high levels of ß-glucosidase, proteolytic and xylanolytic activities were correlated to esters and fatty acids. Our study contributes to a better understanding of the effect of enzymatic activities by yeasts on compound transformations that occur during wine fermentation.

Characterization of the phenolic fraction from Argentine wine and its effect on viability and polysaccharide production of Pediococcus pentosaceus.

OBJECTIVES: To qualitatively and quantitatively characterize a low molecular weight phenolic fraction (LMF) of Malbec wine from Cafayate, Argentina, and evaluate its effect on viability and exopolysaccharide production of Pediococcus pentosaceus 12p, a wine spoilage bacterium. RESULTS: The phenolic compounds detected were, in general, comparable to data previously reported but hydroxycinnamic acids were detected at higher concentrations than determined in other studies. Addition of LMF at identical concentrations present in wine or a four times concentrated LMF mixture to a synthetic wine-like medium produced a diminution in bacterial viability and exopolysaccharide production in the supernatant culture. Transmission electron microscopy revealed damage of bacterial cell integrity after 96 h of incubation only in the presence of four times concentrated LMF. CONCLUSION: This is the first time a low molecular weight phenolic fraction has been characterized in Cafayate wine and it has demonstrated a marked antimicrobial effect on an exopolysaccharide-producing wine spoilage bacterium.

Evaluation of dihydroquercetin-3-O-glucoside from Malbec grapes as copigment of malvidin-3-O-glucoside.

Malbec is a wine grape variety of great phenolic potential characterized for its high levels of anthocyanins and dihydroflavonols. To evaluate the possible implication of dihydroflavonols in the expression of red wine color through reactions of copigmentation or condensation, assays were carried out in wine model systems with different malvidin-3-O-glucoside:dihydroquercetin-3-O-glucoside molar ratios. The addition of increasing levels of dihydroquercetin-3-O-glucoside to a constant malvidin-3-O-glucoside concentration resulted in a hyperchromic effect associated with a darkening of the anthocyanin solutions, greater quantity of color and visual saturation, perceptible to the human eye. Copigmentation and thermodynamic measurements showed that dihydroquercetin-3-O-glucoside can act as an anthocyanin copigment, similar to other usual wine components like flavanols or phenolic acids, although apparently less efficient than flavonols. The high levels of dihydroflavonols existing in Malbec wines in relation to other non-anthocyanin phenolics should make this family of compounds particularly important to explain the color expression in Malbec young red wines.

Phenolic characterisation of red wines from different grape varieties cultivated in Mendoza province (Argentina).

BACKGROUND: Knowledge of the chemical composition of wine and its association with the grape variety/cultivar is of paramount importance in oenology and a necessary tool for marketing. Phenolic compounds are very important quality parameters of wines because of their impact on colour, taste and health properties. The aim of the present work was to study and describe the non-flavonoid and flavonoid composition of wines from the principal red grape varieties cultivated in Mendoza (Argentina). RESULTS: Sixty phenolic compounds, including phenolic acids/derivatives, stilbenes, anthocyanins, flavanols, flavonols and dihydroflavonols, were identified and quantified using high-performance liquid chromatography with diode array detection coupled with electrospray ionisation mass spectrometry (HPLC-DAD/ESI-MS). Marked quantitative differences could be seen in the phenolic profile among varieties, especially in stilbenes, acylated anthocyanins and other flavonoids. CONCLUSION: The polyphenolic content of Malbec wines was higher compared with the other red varieties. Dihydroflavonols represent a significant finding from the chemotaxonomic point of view, especially for Malbec variety. This is the first report on the individual phenolic composition of red wines from Mendoza (Argentina) and suggests that anthocyanins, flavanols and phenolic acids exert a great influence on cultivar-based differentiation.

Phenolic composition of Malbec grape skins and seeds from Valle de Uco (Mendoza, Argentina) during ripening. Effect of cluster thinning.

The phenolic composition of Malbec (Vitis vinifera L.) grape skins and seeds during ripening and the effect of cluster thinning (CT) in two consecutive seasons (2008-2009) were evaluated by high-performance liquid chromatography-diode array detection/electrospray ionization-mass spectrometry (HPLC-DAD/ESI-MS). Removal of 50% of clusters was performed at 40 days (T1), 80 days (T2), and 100 days after flowering (T3) in a vineyard located in southern Mendoza (Argentina). Yield components, with the exception of cluster weight, were significantly affected by CT in both seasons, but no statistically significant differences were found among treatments. Cluster thinning and its timing had little or no influence on physical parameters and fruit chemical composition, and the differences with respect to the control were mainly due to the season. At harvest in 2008, T1 encouraged the biosynthesis of individual anthocyanins in skins, generating 44.0, 39.6, and 41.2% more glucosylated, acetylated, and total anthocyanins, respectively, as compared to the control, whereas in seeds, T1 and T2 mainly changed the concentrations of (+)-catechin, epicatechin-3-gallate, procyanidin B4, dimer gallate 1, trimer gallate 2, and tetramer. Conversely in 2009, T1 significantly affected the content of flavanols and flavonols in skins, whereas in seeds, T1 and T2 modified the level of (+)-catechin, procyanidins B4 and B6, and trimer gallate 2. Moreover, in 2008 the grapes had a higher concentration of most phenolic compounds, indicating a greater potential for more complex wines. Finally, dihydroquercetin-3-glucoside was the major compound among all nonanthocyanin phenolics detected in Malbec skins and represented 25.7% (2008) and 39.9% (2009) of the total content of those compounds at harvest. This finding could represent a distinctive feature of this grape variety.

Optimization of ultrasound assisted-emulsification-dispersive liquid-liquid microextraction by experimental design methodologies for the determination of sulfur compounds in wines by gas chromatography-mass spectrometry.

A new method was developed for analyzing sulfur compounds in the aroma of white wines using ultrasound assisted-emulsification-dispersive liquid-liquid microextraction coupled with gas chromatography-mass spectrometry detection. In the present work, the analytical method for simultaneous determination of seven sulfur compounds (methylmercaptoacetate, methyl(methylthio)acetate, 2-methylthioethanol, 3-methylthiopropanol, 3-methylthiohexanol, 4-methylthio-4-methyl-2-pentanone and hexanethiol) is reported. Parameters that affect the efficiency of the methodology such as extracting and dispersing solvents, sample volume, ion strength, cavitation time and centrifugation time were investigated using a fractionated factorial 2(6-1) (R=V) screening design. Then, the factors presenting significant positive effects on the analytical response (extracting volume, ion strength, cavitation time and centrifugation time) were considered in a further central composite design to optimize the operational conditions for the ultrasound assisted-emulsification-dispersive liquid-liquid microextraction procedure. Additionally, multiple response simultaneous optimization by using the desirability function was used to find the optimum experimental conditions. The best results were obtained using pH sample 4.25, extractant volume 150µL, ionic strength 8.75% NaCl, cavitation time 20s and centrifugation time 50s. The use of the optimized ultrasound assisted-emulsification-dispersive liquid-liquid microextraction technique allowed to obtain the best extraction results with the minimum interference from other substances from the matrix, and it allowed to quantify the analytes in white wine samples by calibration graphs. Recoveries ranging from 91.99% to 125.87% for all sulfur compounds proved the accuracy of the proposed method in white wine samples. Method detection limits were in the range of 0.36-1.67ngmL(-1) and limits of quantitation were between 0.63 and 3.02ngmL(-1) for sulfur compounds in white wine samples. The proposed methodology was successfully applied for the determination concentrations of sulfur compounds in different commercial Chardonnay wine samples from Mendoza, Argentine.

Phenolic characterization of malbec wines from mendoza province (Argentina).

Malbec is a wine grape variety that is now mainly produced in Mendoza and considered to be the emblematic cultivar of Argentina. Forty-four phenolic compounds, including hydroxybenzoic and hydroxycinnamic acids and different flavonoids, were identified and quantified in 61 monovarietal Malbec wines from 11 geographical zones of Mendoza province, using a reversed phase high-performance liquid chromatography system coupled to a diode array detector (HPLC-DAD). Among non-flavonoids, gallic, cis-caftaric, trans-coutaric, and caffeic acids presented the higher concentrations in all of the samples, whereas trans-resveratrol glucoside was present at concentrations from 0.6 to 1.3 mg/L. For the flavonoids, (+)-catechin and (-)-epicatechin presented the higher concentrations among flavan-3-ols with a ratio (+)-catechin/(-)-epicatechin from 1.3 to 2.1. An astilbin derivative and quercetin presented the higher concentrations for flavonols, whereas malvidin-3-glucoside and its derivatives were the major anthocyanins. For the first time the phenolic composition of Malbec wines from Mendoza province has been characterized.