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Aroma is an identifying characteristic of wines and is of crucial importance for the consumer appreciation. Among all odour-active compounds, some key molecules appear only after ageing and give a strong contribution to both identity and added value of the wine bottle. Methyl salicylate (MeSA) is an odorant which usually appears after ageing and could provide an increase in fruity aromas and freshness. This odour-active ester can be found mainly bound to one or more carbohydrates, forming a glycoside. In wines, the hydrolysis of glycosides contributes to the aroma evolution of wine. In our previous works, MeSA was found in relevant content, both in bound and free form, in some genetically related Italian varieties such as Trebbiano di Lugana, Trebbiano di Soave (both employed in the production of Lugana wines), and Verdicchio. In this research, a straightforward filter-and-shot LC-MS/MS method was used for the determination of 7 different MeSA glycosides in 246 samples representative of 18 different monovarietal Italian white wines. MeSA glycosides were found in relevant concentrations in Verdicchio and Lugana wines, whereas they were determined for the first time in all others. Results were in accordance with the literature since Lugana and Verdicchio emerged for the total amount of glycosides, together with Erbaluce, whose richness was not known to date. This trend was observed for all analytes, with stronger intensity, especially in MeSA–glucoside, MeSA–gentiobioside and MeSA–violutoside. The triglycosylated form was not detected in any case. The same trend was also observed for the free form, whose value was not above the olfactory threshold for any sample. MeSA in all its forms also showed a qualitative trend, with MeSA - glucoside and MeSA–primeveroside emphasised in richer cultivars with also a characteristic pattern. The knowledge of the concentration of MeSA glycosides could be considered a potential predictor of the potential evolution of white wines towards balsamic-like nuances.

The initial growth rate of a yeast strain is a key parameter in the production of fermented beverages. Fast growth is linked with higher fermentative capacity and results in less slow and stuck fermentations unable to reach the expected final gravity. As concentrations of metabolites are in a constant state of flux, quantitative data on how growth rate affects the production of aromatic compounds becomes an important factor for brewers. Chemostats allow to set and keep a specific dilution rate throughout the fermentation and are ideal system to study the effect of growth on aroma production. In this study, we ran chemostats alongside batch and fed-batch cultures, compared volatile profiles detected at different growth rates, and identified those affected by the different feeding profiles. Specifically, we quantified six abundant aroma compounds produced in anaerobic glucose-limited continuous cultivations of S. cerevisiae at different dilution rates. We found that volatile production was affected by the growth rate in four out of six compounds assayed, with higher alcohols and esters following opposite trends. Batch and fed-batch fermentations were devised to study the extent by which the final concentration of volatile compounds is influenced by glucose availability. Compared with the batch system, fed-batch fermentations, where the yeast growth was artificially limited by a slow constant release of nutrients in the media, resulted in a significant increase in concentration of higher alcohols, mirroring the results obtained in continuous fermentations. This study paves the way to further process development optimization for the production of fermented beverages.

Thanks to their low odor detection thresholds, free varietal thiols (VTs) play a key role in the primary aroma of wines, to which they confer an intense scent reminiscent of box tree, grapefruit, citrus fruits, passionfruit and cat urine odor. Excluding wines from a few VT-rich grapevine cultivars, VTs appear to be present in most cultivars at trace levels, although a comprehensive dataset is still missing. The low concentration of VTs combined with their high reactivity and matrix complexity make their determination in wines a challenging task. In this research an optimized liquid chromatography – tandem mass spectrometry (LC-MS/MS) method was validated and used for the quantification of 4-methyl-4-sulfanylpentan-2-one (4-MSP), 3-sulfanylhexan-1-ol (3-SH), 3-sulfanylhexyl acetate (3-SHA) and ethyl 3-sulfanylpropionate (E3SP) in 246 samples (vintage 2019) representative of 18 monovarietal Italian white wines. VTs were detected in all cultivars even though higher values of 3-SH were found in Lugana, Müller-Thurgau and Verdicchio cultivars. Müller-Thurgau wines showed the highest level of 4- MSP, that was mainly correlated to the odor descriptors of passionfruit and box tree/cat urine. The VTs composition of Müller-Thurgau was confirmed on a second set of 50 wines from different vintages. From a sensory perspective, the samples of Müller-Thurgau showed the best positive correlations between chemical variables and the odor descriptors thiol note, passion fruit and box tree/cat urine. These notes are significantly related to 4-MSP, suggesting that it could play a relevant olfactory role for the aroma of Müller-Thurgau wines. Sorting analysis allowed to group these wines according to their thiolic characteristics. The chemical variables and the odor descriptors attributable to the thiol notes are important for Müller-Thurgau and Lugana wines, while the contribution of thiol notes was sensorially negligible for the other wines.

Phyto-thermotherapy is a treatment consisting in immersing oneself in baths of self-heating alpine grass, to benefit of the heat and rich aromatic components released by the process. The aim of this study was to characterize the bacterial and fungal diversity of three phyto-thermal baths (PTB) performed in three different months, and to compare the data with the profile of the volatile organic compounds (VOCs) of the process. All the data collected showed that PTBs were structured in two stages: the first three days were characterised by an exponential rise of the temperature, a fast bacterial development, higher microbial diversity and higher concentrations of plant aliphatic hydrocarbons. The second stage was characterised by a stable high temperature, shrinkage of the microbial diversity with a predominance of few bacterial and fungi species and higher concentrations of volatiles of microbial origin. Erwinia was the dominant microbial species during the first stage and probably responsible of the self-heating process. In conclusion, PTBs has shown both similarities with common self-heating processes and important peculiarities such as the absence of pathogenic bacteria and the dominance of plant terpenoids with health characteristics among the VOCs confirming the evidence of beneficial effects in particular in the first three days.

Aim. We aimed to study the impact of water deficit on the concentration of key flavour and phenolic secondary metabolites of wines.Methods and results. A drought-stress field trial was conducted on Vitis vinifera cv. Merlot and Tocai Friulano for two seasons. Fully irrigated (C) and deficit irrigated (D) grapes were microvinified and the resulting wines were analysed to determine the concentrations of anthocyanins, tannins, and free and glycosidically-bound Volatile Organic Compounds (VOCs). A descriptive sensory test was undertaken on the same wines. Water stressed grapes produced wines with higher concentrations of anthocyanins in Merlot and of free and glycosidically-bound monoterpenes in Tocai Friulano. Both cultivars displayed higher amounts of glycosidically-bound C13-norisoprenoids.Conclusions. Previously observed drought-induced compositional changes to the grapes were transfered to the wines, with an increase in polyphenols and VOCs. However, the timing and the duration of the water stress in the field only heavily impacted the final wine composition with major metabolic modification when the severe water deficit started early (at approximately 40 days after anthesis) and lasted over the entire season until harvest.Significance and impact of the study. This study highlights the positive role of a controlled water deficit on the composition of the wines in terms of secondary metabolites.

The influence of grape maturity on wine volatome was investigated using HS-SPME-GC × GC-TOFMS. Shiraz wines were made from grapes harvested from four different vineyards from two berry maturity levels. A total of 1276 putative compounds were detected in at least one of the wine samples and 175 showed significant trends related to grape maturity. The first two dimensions of the Principal component analysis accounted for 57% of the variation and separated the samples according to the harvest date. Wines from the first harvest date were characterised by an abundance of lipoxygenase derived compounds, norisoprenoids and sulfur-containing compounds whereas a significant increase in some acetate esters was observed in wines produced from the more mature grapes. This study demonstrated a common evolution of grape volatiles for Shiraz inside the same mesoclimate. During the late ripening stage of the grape, a direct nexus between sugar concentration and wine volatile evolution was not observed.

White wines from two vintages were bottled in green and flint glass bottles and stored under typical supermarket shelf conditions. Light-strike fault, colour changes, light and temperature exposure were monitored up to 50 days. In general, green glass bottles secured wine quality for the tested period. Only a few flint glass bottled wines developed a fault due to light-strike after 1–2 days of supermarket shelf life, but all developed the fault after 20–40 days; so we considered the possibility of two mechanisms causing the problem. Flint-glass bottled wines stored on the bottom shelf developed the defect only slightly later compared with the wines on the top shelf. Moreover, storing the wines in dark and cold after a period of exposure to light did not eliminate the fault, indicating that the defect is probably additive. Monitoring the amount of light to which wines are exposed could prevent the fault.

To differentiate white wines from Croatian indigenous varieties, volatile aroma compounds were isolated by headspace solid-phase microextraction (HS-SPME) and analyzed by comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC&times, GC-TOF-MS) and conventional one-dimensional GC-MS. The data obtained were subjected to uni- and multivariate statistical analysis. The extra separation ability of the GC&times, GC second dimension provided additional in-depth volatile profile information, with more than 1000 compounds detected, while 350 were identified or tentatively identified in total by both techniques, which allowed highly efficient differentiation. A hundred and sixty one compounds in total were significantly different across monovarietal wines. Monoterpenic compounds, especially &alpha, terpineol, followed by limonene and linalool, emerged as the most powerful differentiators, although particular compounds from other chemical classes were also shown to have notable discriminating ability. In general, &Scaron, krlet wine was the most abundant in monoterpenes, Malvazija istarska was dominant in terms of fermentation esters concentration, Po&scaron, ip contained the highest levels of particular C13-norisoprenoids, benzenoids, acetates, and sulfur containing compounds, Kraljevina was characterized by the highest concentration of a tentatively identified terpene &gamma, dehydro-ar-himachalene, while Mara&scaron, tina wine did not have specific unambiguous markers. The presented approach could be practically applied to improve defining, understanding, managing, and marketing varietal typicity of monovarietal wines.

Processing citrus fruits into juice generates large amounts of by-products, mainly pomaces. This study aimed to perform a comprehensive analysis of the composition in phenolic compounds, carotenoids, and volatile organic compounds (VOCs) of juices and pomaces of four citrus fruits from Southern Italy, i.e., mandarin, lemon, orange, and bergamot. Juices were produced by squeezing the fruits into an electrical juicer, whereas pomaces were obtained as by-products of the juice extraction. The phytochemical content of the samples was studied by targeted LC and GC approaches. Results indicated that lemon provided the juice with the greatest phenolic content. It was abundant in eriocitrin (90.9 ± 10.8 mg kg−1 FW), isorhamnetin 3-O-rutinoside (47.3 ± 8.03 mg kg−1 FW), and rutin (78.9 ± 14.5 mg kg−1 FW). Likewise, lemon pomace was the richest in phenolics, mostly narirutin (130 ± 14.7 mg kg−1 FW). As regards carotenoids, mandarin and orange pomaces were equally (p > 0.05) prominent sources of the compounds, providing primarily lutein and β-cryptoxanthin. The phytochemical profile of lemon and mandarin pomaces was unknown up to date. Bergamot accumulated great amounts of VOCs. In particular, bergamot juice was rich in monoterpenes, e.g., α-pinene (375 ± 62.7 mg kg−1 FW) and γ-terpinene (551 ± 67 mg kg−1 FW). The study investigated for the first time the carotenoid and VOCs profiles of bergamot products, and of mandarin and lemon pomaces. Since, citrus pomaces contained great amounts of phytochemicals, they should find new applications in the food and cosmetic industries.

Background and Aims This study evaluated the impact that the non-Saccharomyces yeasts, Torulaspora delbrueckii (TD), Lachancea thermotolerans (LT), Pichia kluyveri (PK), Metschnikowia pulcherrima (MP), Candida zemplinina (CZ) and Kazachstania aerobia (KA), in sequential inoculation with Saccharomyces cerevisiae (SC), had on the volatile chemical profile of Shiraz wine. Methods and Results Shiraz grape must was inoculated with monocultures of the non-Saccharomyces yeasts, which were allowed to ferment until 2% v/v ethanol concentration was reached at which point, SC was added to complete the alcoholic fermentation. The control was SC in monoculture. The final wines were subjected to solid phase microextraction-GC × GC-time of flight-MS to evaluate the untargeted volatile metabolite profile of each treatment. Each fermentation produced a unique chemical profile. The LT–SC sequential fermentation was the most significantly different from the control primarily in the ester, alcohol and terpene profile. The KA–SC sequential fermentation had the highest amount of volatile acidity, and the PK–SC sequential fermentation had a relatively high amount of acetaldehyde and a few esters. The MP–SC sequential fermentation also revealed a higher concentration of several esters. The TD–SC sequential fermentation was notable for its lack of a distinct pattern in comparison with that of the other fermentations. Conclusions Given these characteristics, the LT–SC sequential fermentation showed the most potential for increased chemical complexity of the Shiraz volatile profile. Significance of the Study The results demonstrate that there is no clear, singular trend for how different, non-Saccharomyces species of yeast – once thought to be wholly unfavourable in wine fermentations – will impact wine chemistry, flavour or quality. Each species presents unique metabolic characteristics, many of which could be beneficial, and their potential application in the wine industry should be considered.

Fungi produce a variety of volatile organic compounds (VOCs) during their primary and secondary metabolism. In the beverage industry, these volatiles contribute to the the flavor and aroma profile of the final products. We evaluated the fermentation ability and aroma profiles of non-conventional yeasts that have been associated with various food sources. A total of 60 strains were analyzed with regard to their fermentation and flavor profile. Species belonging to the genera Candida, Pichia and Wickerhamomyces separated best from lager yeast strains according to a principal component analysis taking alcohol and ester production into account. The speed of fermentation and sugar utilization were analysed for these strains. Volatile aroma-compound formation was assayed via gas chromatography. Several strains produced substantially higher amounts of aroma alcohols and esters compared to the lager yeast strain Weihenstephan 34/70. Consequently, co-fermentation of this lager yeast strain with a Wickerhamomyces anomalus strain generated an increased fruity-flavour profile. This demonstrates that mixed fermentations utilizing non-Saccharomyces cerevisiae biodiversity can enhance the flavour profiles of fermented beverages.

Every grape cultivar has its own unique genetic characteristics, leading to the production of a different secondary metabolite profile. Aroma is one of the most important aspects in terms of the quality of grapes and previous studies have assigned specific aromas to particular grape cultivars. In this study we present the molecular profiling of volatile aroma metabolites and their precursors in ten selected genotypes, including six Vitis vinifera cultivars, two American species (Arizonica Texas, Vitis cinerea) and two interspecific crosses. Chemical profiling was achieved through combined use of two orthogonal techniques, GC-MS and LC-HRMS, before and after enzymatic hydrolysis. The results show that both free and glycosidically bound aroma precursors behave differently in each different grape cultivar and species. As many as 66 free aroma volatile molecules (originally existing and released after hydrolysis) were profiled through GC-MS analysis, while 15 glycosylated precursors of volatiles were identified through LC-HRMS and correlation with GC-MS data.

Saccharomyces cerevisiae is the main driver of alcoholic fermentation. It is typically inoculated at high levels to ensure successful implantation as well as reduce the risks of stuck fermentations and off-flavor production. However, winemakers have found that wines produced with only S. cerevisiae can be lacking in complexity compared to fermentations where non-Saccharomyces yeasts are more active. This study sought to understand the early fermentation characteristics of Kazachstania gamospora, Lachancea thermotolerans, Metschnikowia pulcherrima, Torulaspora delbrueckii and Zygosaccharomyces kombuchaensis in both Sauvignon blanc and Syrah musts. S. cerevisiae was used as a control. Solid-phase microextraction coupled to GC–MS was used to evaluate the musts once they reached 2% ethanol concentration. The method targeted 90 different compounds known to occur in wine and/or be produced by yeast during fermentation. For the first time, K. gamospora and Z. kombuchaensis have been studied in the context of wine. While the other yeasts are commercially available starter cultures, they have never been profiled this extensively. Analysis showed that each yeast profile was unique and different based on the must. The non-Saccharomyces yeasts produced lower concentrations of esters, alcohols and terpenes with the exception of K. gamospora which produced more total esters than the control.

Glycosides are ubiquitous plant secondary metabolites consisting of a non-sugar component called an aglycone, attached to one or more sugars. One of the most interesting aglycones in grapes and wine is methyl salicylate (MeSA), an organic ester naturally produced by many plants, particularly wintergreens. To date, nine different MeSA glycosides from plants have been reported, mainly spread over the genera Gaultheria, Camellia, Polygala, Filipendula, and Passiflora. From a sensorial point of view, MeSA has a balsamic-sweet odor, known as Wintergreen. MeSA was found in Vitis riparia grapes, in Vitis vinifera sp. and in the Frontenac interspecific hybrid. We found that the MeSA glycosides content in Verdicchio wines and in some genetically related varieties (Trebbiano di Soave and Trebbiano di Lugana) was very high. In order to understand which glycosides were present in wine, the methanolic extract of Verdicchio wine was injected into a UPLC-Q-TOF-HDMS and compared to the extracts of different plants rich in such glycosides. Using pure standards, we confirmed the existence of two glycosides in wine: MeSA 2-O--d-glucoside and MeSA 2-O--d-xylopyranosyl (1-6) -d-glucopyranoside (gaultherin). For the first time, we also tentatively identified other diglycosides in wine: MeSA 2-O--l-arabinopyranosyl (1-6)--d-glucopyranoside (violutoside) and MeSA 2-O--d-apiofuranosyl (1-6)--d-glucopyranoside (canthoside A), MeSA 2-O--d-glucopyranosyl (1-6)-O--d-glucopyranoside (gentiobioside) and MeSA 2-O--l-rhamnopyranosyl (1-6)--d-glucopyranoside (rutinoside). Some of these glycosides have been isolated from Gaultheria procumbens leaves by preparative liquid chromatography and structurally annotated by 1H- and 13C-NMR analysis. Two of the peaks isolated from Gaultheria procumbens leaves, namely MeSA sambubioside and MeSA sophoroside, were herein observed for the first time. Six MeSA glycosides were quantified in 64 Italian white wines, highlighting the peculiar content and pattern in Verdicchio wines and related cultivars. The total concentration in bound and free MeSA in Verdicchio wines varied in the range of 456&ndash, 9796 g/L and 5.5&ndash, 143 g/L, respectively, while in the other wines the bound and free MeSA was below 363 g/L and 12 g/L, respectively. As this compound&rsquo, s olfactory threshold is between 50 and 100 g/L, our data support the hypothesis that methyl salicylate can contribute to the balsamic scent, especially in old Verdicchio wines.

Saccharomyces cerevisiae (SC) is the main driver of alcoholic fermentation, however for aroma and flavor formation in wine, non-Saccharomyces species can have a powerful effect. This study aimed to compare untargeted volatile compound profiles from SPME–GCxGC-TOF-MS and sensory analysis data of Sauvignon blanc wine inoculated with six different non-Saccharomyces yeasts followed by SC. Torulaspora delbrueckii (TD), Lachancea thermotolerans (LT), Pichia kluyveri (PK) and Metschnikowia pulcherrima (MP) where commercial starter strains, while Candida zemplinina (CZ) and Kazachstania aerobia (KA), were isolated from wine grape environments. Each wine showed a distinct profile both sensorially and chemically. SC and CZ wines were the most distinct in both of these cases. SC wine had guava, grapefruit, banana, and pineapple aromas while CZ wine was driven by fermented apple, dried peach/apricot, and stewed fruit as well as sour flavor. Chemically over 300 unique features were identified as significantly different across the fermentations. SC wine had the highest number of esters in the highest relative concentration but all the yeasts had distinct ester profiles. CZ wine displayed the highest number of terpenes in high concentration but also produced a large amount of acetic acid. KA wine was high in ethyl acetate. TD wine had fewer esters but three distinctly higher thiol compounds. LT wine showed a relatively high number of increased acetate esters and certain terpenes. PK wine had some off odor compounds while the MP wine had high levels of methyl butyl-, methyl propyl-, and phenethyl esters. Overall, this study gives a more detailed profile of these yeasts contribution to Sauvignon blanc wine than previously reported.

Biosynthesis of the sesquiterpene rotundone in Vespolina grapes during berry ripening was investigated over two consecutive seasons, revealing that the compound accumulates from veraison to harvest and reaches relatively high concentrations (up to 5.44 μg/kg). Rotundone levels up to 1.91 μg/kg were also found in clones of Gruener Veltliner, a white grape variety known to give 'peppery' wines. These concentrations are higher than those reported for Syrah grapes and are similar to the levels found in some plants. Rotundone was shown to accumulate almost exclusively in berry exocarp, suggesting that skin contact during winemaking could be used to modulate the peppery character of red wine. However, rotundone yield after the winemaking process was relatively low. Indeed, only 10% of the rotundone present in grapes was extracted during fermentation, and only 6% was recovered in bottled wine. The results presented in this work provide key knowledge for manipulation of the peppery character of wine in order to optimize the intensity of this characteristic wine aroma.

Supercritical pasteurisation is receiving increasing attention as an alternative technology for foodstuff pasteurisation, but often the possible effects on the perceptible quality are not sufficiently considered. To address this latter issue, besides standard microbial analysis, we here investigate the impact of CO 2 /N 2 O supercritical pasteurisation (100 bar, 36 °C and 10 min treatment time) on the quality traits of fresh apple juice, linked to consumer perception. Discriminative sensory analysis (triangle test) and basic chemical characterization (total solids, sugars, organic acids, polyphenols) could not clearly demonstrate any induced modification of the treated juice, while head space analysis of volatile compounds (both by GC–MS and PTR–MS) indicated a general depletion of the volatile compounds that must be considered in the development of a stabilization method based on supercritical gases.

We carried out comprehensive mapping of volatile compounds in 70 wines, from 48 wineries and 6 vintages, representative of the two main production areas for Italian sparkling wines, by HS-SPME-GCxGC-TOF-MS and multivariate analysis. The final scope was to describe the metabolomics space of these wines, and to verify whether the grape cultivar signature, the pedoclimatic influence of the production area, and the complex technology were measurable in the final product. The wine chromatograms provided a wealth of information, with 1695 compounds being found. A large number of putative markers influenced by the cultivation area was observed. A subset of 196 biomarkers fully discriminated between the two types of sparkling wines investigated. Among the new compounds, safranal and α-isophorone were observed. We showed how correlation-based network analysis could be used as a tool to detect the differences in compound behaviour based on external/environmental influences.

The lactic acid populations of 2 seasonal Puzzone di Moena cheeses made from winter and summer raw cow's milk were characterized at different ripening times. Lactic acid bacteria (LAB) were isolated on selective media and subjected to genetic typing and identification. The species most frequently found during ripening were Lactobacillus paracasei ssp. paracasei, Lactobacillus plantarum, and Pediococcus pentosaceus. The different strains recognized by random amplification of polymorphic DNA-PCR were characterized for their acidifying and proteolytic activities to select nonstarter LAB to be used as secondary adjunct cultures (SAC). For each of the 3 above species, a strain showing weak acidification and high proteolytic capacity was selected. The 3 strains (Lb. paracasei ssp. paracasei P397, Lb. plantarum P399, and P. pentosaceus P41) constituted a mixed SAC used at 2 levels of concentration (10(3) and 10(4) cfu/mL) in experimental cheese making at dairy factory-scale. The analysis of volatile organic compounds as well as sensory analyses showed that the preferred level of SAC inoculation was 10(3) cfu/mL.