Quantitation of trace polyfunctional thiols in wine by liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry in parallel reaction monitoring.

Polyfunctional thiols are key contributors to wine aroma due to their extremely low odor thresholds, and their quantitative analysis remains challenging as a result of their ultratrace concentrations and high reactivity. This work presents the first method based on ultra-high-performance liquid chromatography (UHPLC) coupled to quadrupole Orbitrap high-resolution mass spectrometry (HRMS) in parallel reaction monitoring (PRM) mode for quantifying thiols at nanograms per liter (ng/L) levels in wine. Thiols in wine were derivatized using 4,4'-dithiodipyridine and isolated by liquid-liquid extraction. This protocol allowed rapid sample preparation with minimum labor input and low consumable expenses. Instrumental analysis was conducted using UHPLC-quadrupole Orbitrap HRMS in PRM mode. Twenty thiol analytes, including literature-known, recently identified, and novel thiols were selected and validated by the optimized method in three wine matrices. The overall analytical performances demonstrated by this method were equivalent, and in most cases, greater than many previously developed GC-MS or LC-MS methods. The validated method was applied to analyze a selection of wines in which 12 thiols were quantified.

Identification, Quantitation, and Sensory Evaluation of Thiols in Bordeaux Red Wine with Characteristic Aging Bouquet.

Great Bordeaux red wines are known for their distinctive aging bouquet. However, the nature of volatile chemicals underpinning this sensory expression is not fully understood. This work investigated the empyreumatic aging bouquet of a collection of premium Bordeaux red wines using silver-ion (Ag+) solid-phase extraction, cryogenic heart-cutting multidimensional gas chromatography mass spectrometry/olfactometry, and comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry. In doing so, a substantial number of "meaty" odors were revealed. Three detected "meaty" notes were tentatively or unequivocally attributed to furan thiols. Among them, 2-methyltetrahydrofuran-3-thiol (1) with a pleasant "meaty" aroma was reported in wine for the first time. Its trans isomer (trans-1a) was resolved from its racemate by chemical modification, which confirmed its presence in wine. The odor detection threshold of trans-1a in the model wine was determined at 55 ng/L. Moreover, an additive effect between 1 and literature-known 2-methyl-3-furanthiol was observed. By a new ultra high-performance liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry method, the concentration of trans-1a, in addition to those of 2-methyl-3-furanthiol and 2-furfuryl thiol, was measured in the wines at ng/L levels.

Impact of the Closure Oxygen Transfer Rate on Volatile Compound Composition and Oxidation Aroma Intensity of Merlot and Cabernet Sauvignon Blend: A 10 Year Study.

This study evaluated the impact of closure type on unoaked 100 %-Merlot, oak-aged 70%-Merlot/30%-Cabernet Sauvignon, and 30%-Merlot/70%-Cabernet Sauvignon during a 10 year period. Closures were microagglomerate corks, screw caps, and synthetics with the known oxygen transfer rate (OTR), ranging from 0.1 to 4.6 mg/y, including natural corks. Oxidation intensity perception, dissolved oxygen, sulfite, and 3-methyl-2,4-nonanedione (MND) were monitored on a regular basis. After 10 years of aging, additional aroma impact markers were evaluated (3-sulfanylhexan-1-ol, H2S, DMS, methional, and phenylacetaldehyde). Low OTR levels (≤0.3 mg/y) delayed the oxidation of red wines in this long-term experiment. In addition, our results led us to hypothesize that the MND concentration in young wines might be linked with their ability to produce it during bottle aging that is with their aging potential. Finally, we found that the kinetic accumulation of MND in wines was first strongly impacted by its intrinsic composition and thereafter by the OTRT0 of the stopper.

Methyl Salicylate, an Odor-Active Compound in Bordeaux Red Wines Produced without Sulfites Addition.

Red wines produced without the addition of any SO2 are currently the source of a new consumer trend. The first characterization approaches regarding these specific wines were devoted to sensory studies that highlighted differences according to the use of SO2 during winemaking. The goal of this paper is to extend our knowledge of such aromatic specificities. Examining experimental wines produced with and without the addition of SO2, aroma fractionation, gas chromatography coupled with olfactometry, and mass spectrometry were applied to identify compounds at the origin of the specificities of these wines. Thus, we identified methyl salicylate as being impacted by the use of SO2 during the winemaking process. Studying a large number of commercial Bordeaux red wines, methyl salicylate was significantly quantified at a higher content in wines without added SO2. A sensory approach revealed a significant impact of methyl salicylate on wines without the sulfite aroma, particularly concerning fruity aromas and wine freshness.

Identification and Distribution of New Impact Aldehydes in Toasted Oak Wood (Quercus petraea).

The purpose of this study was to identify aroma compounds associated with the toasting intensity of oak wood (Quercus petraea). Crude organic extracts from oak wood samples (toasted at different temperature-time couples) were analyzed by a sensory-guided approach using GC-O-TOFMS, followed by purification with semipreparative HPLC (reverse phase). This approach revealed two specific odorous zones (OZs) reminiscent of metal and puff pastry. The first OZ was identified as trans-4,5-epoxy-(E)-2-decenal (1) by coinjection of the commercial product, whereas identification of (2E,4E,6Z)-nonatrienal (2) associated with puff pastry OZ was validated by a multistep chemical synthesis approach (Wittig reaction) followed by semipreparative HPLC purification (chiral phase). Their detection thresholds in model wine solution were 60 ng/L (1) and 16 ng/L (2). Their distribution in toasted oak wood samples [GC-NCI-MS (NH3) analysis] ranged from some ng/g to 210 ng/g for (1) and 85 ng/g for (2). Finally, additional sensory experiments demonstrated the impact of newly identified aldehydes in toasted oak wood.

Qualitative Screening of Volatile Thiols in Wine by Selective Silver Ion Solid-Phase Extraction with Heart-Cutting Multidimensional Gas Chromatography Mass Spectrometry/Olfactometry.

The chemical analysis of odorous volatile thiols is intrinsically challenging. Substantial progress has been made in quantitative analysis of targeted thiols at ultra-trace concentrations (ng/L), but lesser analytical attention has been given to the qualitative screening of unknown thiols. This work presents a solid-phase extraction (SPE) method using a silver ion (Ag+)-based sorbent to isolate volatile thiols from red wine. This proposed Ag+ SPE method was effective (recovery: 87-101% for four non-furan thiols and 35-49% for two furan thiols), simple, safe, and greatly reduced artifacts, testifying to its suitability as the sample preparation protocol for a qualitative screening experiment. Separation and detection were conducted using heart-cutting multidimensional gas chromatography coupled to mass spectrometry/olfactometry (H/C MDGC-MS/O). Key parameters including H/C width, main host oven temperature, and cryogenic trapping temperature were investigated for optimal instrument performance. The developed Ag+ SPE H/C MDGC-MS/O strategy was readily applicable for qualitative screening of odorous volatile thiols in wine, as demonstrated by two case studies.

Modifications of Grapevine Berry Composition Induced by Main Viral and Fungal Pathogens in a Climate Change Scenario.

The grapevine is subject to high number of fungal and viral diseases, which are responsible for important economic losses in the global wine sector every year. These pathogens deteriorate grapevine berry quality either directly via the modulation of fruit metabolic pathways and the production of endogenous compounds associated with bad taste and/or flavor, or indirectly via their impact on vine physiology. The most common and devastating fungal diseases in viticulture are gray mold, downy mildew (DM), and powdery mildew (PM), caused, respectively by Botrytis cinerea, Plasmopara viticola, and Erysiphe necator. Whereas B. cinerea mainly infects and deteriorates the ripening fruit directly, deteriorations by DM and PM are mostly indirect via a reduction of photosynthetic leaf area. Nevertheless, mildews can also infect berries at certain developmental stages and directly alter fruit quality via the biosynthesis of unpleasant flavor compounds that impair ultimate wine quality. The grapevine is furthermore host of a wide range of viruses that reduce vine longevity, productivity and berry quality in different ways. The most widespread virus-related diseases, that are known nowadays, are Grapevine Leafroll Disease (GLRD), Grapevine Fanleaf Disease (GFLD), and the more recently characterized grapevine red blotch disease (GRBD). Future climatic conditions are creating a more favorable environment for the proliferation of most virus-insect vectors, so the spread of virus-related diseases is expected to increase in most wine-growing regions. However, the impact of climate change on the evolution of fungal disease pressure will be variable and depending on region and pathogen, with mildews remaining certainly the major phytosanitary threat in most regions because their development rate is to a large extent temperature-driven. This paper aims to provide a review of published literature on most important grapevine fungal and viral pathogens and their impact on grape berry physiology and quality. Our overview of the published literature highlights gaps in our understanding of plant-pathogen interactions, which are valuable for conceiving future research programs dealing with the different pathogens and their impacts on grapevine berry quality and metabolism.

Impact of Closure OTR on the Volatile Compound Composition and Oxidation Aroma Intensity of Sauvignon Blanc Wines during and after 10 Years of Bottle Storage.

The oxygen transfer rate (OTR) of closures is a well-known parameter impacting the quality of Sauvignon blanc wines (SBw) within the first years of storage, but little research has been published on its long-term effects. The chemical changes in oxidation odor intensity in three SBw sealed with natural cork and other closures that had different known OTRs, ranging from <0.1 to 4.6 mg/year, were monitored over a 10 year period. During aging, free SO2 and 3-sulfanylhexanol loss, concomitant with increases in dissolved O2, OD420, and sotolon, were correlated with closure OTR levels. After 10 years of aging, sensory analysis was conducted, supported by additional chemical analysis of aroma impact markers, including methional, phenylacetaldehyde, 2-furanmethanthiol, 4-methyl-4-sulfanylpentan-2-one, ethyl-2-sulfanylacetate, and hydrogen sulfide, as well as total SO2 and dissolved CO2. These analyses revealed that selected SBw were protected from oxidation over a 10 year aging period, provided that the closure OTR did not exceed 0.3 mg/year.

Strategies for the identification and sensory evaluation of volatile constituents in wine.

Wine aroma, which stems from complex perceptual and cognitive processes, is initially driven by a multitude of naturally occurring volatile constituents. Its interpretation depends on the characterization of relevant volatile constituents. With large numbers of volatile constituents already identified, the search for unknown volatiles in wine has become increasingly challenging. However, the opportunities to discover unknown volatile compounds contributing to the wine volatilome are still of great interest, as demonstrated by the recent identification of highly odorous trace (µg/L) to ultra-trace (ng/L) volatile compounds in wine. This review provides an overview of both existing strategies and future directions on identifying unknown volatile constituents in wine. Chemical identification, including sample extraction, fractionation, gas chromatography, olfactometry, and mass spectrometry, is comprehensively covered. In addition, this review also focuses on aspects related to sensory-guided wine selection, authentic reference standards, artifacts and interferences, and the evaluation of the sensory significance of discovered wine volatiles. Powerful key volatile odorants present at ultra-trace levels, for which these analytical approaches have been successfully applied, are discussed. Research areas where novel wine volatiles are likely to be identified are pointed out. The importance of perceptual interaction phenomena is emphasized. Finally, future avenues for the exploration of yet unknown wine volatiles by coupling analytical approaches and sensory evaluation are suggested.

Sucrose Metabolism and Transport in Grapevines, with Emphasis on Berries and Leaves, and Insights Gained from a Cross-Species Comparison.

In grapevines, as in other plants, sucrose and its constituents glucose and fructose are fundamentally important and carry out a multitude of roles. The aims of this review are three-fold. First, to provide a summary of the metabolism and transport of sucrose in grapevines, together with new insights and interpretations. Second, to stress the importance of considering the compartmentation of metabolism. Third, to outline the key role of acid invertase in osmoregulation associated with sucrose metabolism and transport in plants.

Contribution of Grapes and Oak Wood Barrels to Pyrrole Contents in Chardonnay Wines: The Influence of Several Cooperage Parameters.

The influence of some enological parameters on pyrrole concentrations in Chardonnay wines was studied. First, a quantitative method to assay five pyrroles was optimized and applied to determine their content in wines produced in different containers. All pyrroles were observed in wines aged in a stainless-steel tank, which indicated that they have a varietal or fermentative origin. However, their concentrations were significantly higher in wines made in new barrels than in older barrels or in a stainless-steel tank, so oak wood may largely contribute during the winemaking process. A quantitative method to assay pyrroles in oak wood extract was also developed to study the influence of several cooperage parameters such as different types of traditional toasting, as well as the temperature and the time of toasting. Significant differences were observed on pyrrole concentrations in oak wood extracts according to these different cooperage parameters. These findings bring new perspectives to the monitoring of winemaking and the aging of Chardonnay wines.

Methyl salicylate, a grape and wine chemical marker and sensory contributor in wines elaborated from grapes affected or not by cryptogamic diseases.

Methyl salicylate (MeSA) is a plant metabolite that induces plant defence resistance and an odorous volatile compound presenting green nuances. This volatile compound was shown to be present in wine samples, sometimes at concentrations above its olfactory detection threshold. MeSA is localized in grapes, particularly in the skins and stems, and is extracted during red wine vinification. It was detected at the highest concentrations in wines of several grape varieties, made from grapes affected by cryptogamic diseases, namely downy mildew caused by Plasmopara viticola, and black rot caused by Guignardia bidwellii. It has also been detected in wines from vines affected by Esca, a Grapevine Trunk Disease. MeSA can also be considered to be a chemical marker in grapes and wine indicative of the level of development of several vine cryptogamic diseases.

Sensorial Impact and Distribution of 3-Methyl-2,4-nonanedione in Cognacs and Spirits.

The aim of this study was to improve our knowledge on the chemical markers of Cognac aromas. We report results concerning the distribution and sensorial impact of 3-methyl-2,4-nonanedione (MND), a well-known compound in aged red wine, reminiscent of anise or "dried fruit", according to its concentration. We assayed first this diketone (solid-phase microextraction (SPME)-gas chromatography (GC)/mass spectrometry (MS), chemical ionization (CI)) in many Cognac samples followed by grappa, brandy, rum, whisky, vodka, and fruit spirits, and concentrations ranged from traces to 11.2 µg/L. Highest concentrations were obtained in grappa and freshly distilled eaux-de-vie of Cognac samples. Exceeding its detection threshold (100 ng/L, 70 vol %), MND contributes to the anise descriptor of these spirits. Its concentration decreased over aging while being highly correlated with the total amount of fatty acid ethyl ester. In addition, we showed that MND was produced during distillation according to the oxidation state of the white wine as well as the amount of lees used.

Grape Berry Secondary Metabolites and Their Modulation by Abiotic Factors in a Climate Change Scenario-A Review.

Temperature, water, solar radiation, and atmospheric CO2 concentration are the main abiotic factors that are changing in the course of global warming. These abiotic factors govern the synthesis and degradation of primary (sugars, amino acids, organic acids, etc.) and secondary (phenolic and volatile flavor compounds and their precursors) metabolites directly, via the regulation of their biosynthetic pathways, or indirectly, via their effects on vine physiology and phenology. Several hundred secondary metabolites have been identified in the grape berry. Their biosynthesis and degradation have been characterized and have been shown to occur during different developmental stages of the berry. The understanding of how the different abiotic factors modulate secondary metabolism and thus berry quality is of crucial importance for breeders and growers to develop plant material and viticultural practices to maintain high-quality fruit and wine production in the context of global warming. Here, we review the main secondary metabolites of the grape berry, their biosynthesis, and how their accumulation and degradation is influenced by abiotic factors. The first part of the review provides an update on structure, biosynthesis, and degradation of phenolic compounds (flavonoids and non-flavonoids) and major aroma compounds (terpenes, thiols, methoxypyrazines, and C13 norisoprenoids). The second part gives an update on the influence of abiotic factors, such as water availability, temperature, radiation, and CO2 concentration, on berry secondary metabolism. At the end of the paper, we raise some critical questions regarding intracluster berry heterogeneity and dilution effects and how the sampling strategy can impact the outcome of studies on the grapevine berry response to abiotic factors.

Biosynthesis and Cellular Functions of Tartaric Acid in Grapevines.

Tartaric acid (TA) is an obscure end point to the catabolism of ascorbic acid (Asc). Here, it is proposed as a "specialized primary metabolite", originating from carbohydrate metabolism but with restricted distribution within the plant kingdom and lack of known function in primary metabolic pathways. Grapes fall into the list of high TA-accumulators, with biosynthesis occurring in both leaf and berry. Very little is known of the TA biosynthetic pathway enzymes in any plant species, although recently some progress has been made in this space. New technologies in grapevine research such as the development of global co-expression network analysis tools and genome-wide association studies, should enable more rapid progress. There is also a lack of information regarding roles for this organic acid in plant metabolism. Therefore this review aims to briefly summarize current knowledge about the key intermediates and enzymes of TA biosynthesis in grapes and the regulation of its precursor, ascorbate, followed by speculative discussion around the potential roles of TA based on current knowledge of Asc metabolism, TA biosynthetic enzymes and other aspects of fruit metabolism.

Sensory-directed characterisation of distinctive aromas of Sauternes and Viognier wines through semi-preparative liquid chromatography and gas chromatography approaches.

Gas chromatography-olfactometry-mass spectrometry (GC-O-MS) has been very useful in identifying aroma compounds from within the complex matrix of wine. Supplementary separation can be required to overcome co-elution of volatiles or other sensory-directed chromatographic strategies are needed, including multidimensional chromatography and preparative fraction collection coupled to GC. Studies investigating 'overripe orange' aroma in sweet Sauternes wine and the similar 'apricot' aroma in Viognier wine were conducted. Wines with the targeted aroma attributes were selected and concentrated wine extracts prepared. GC-O found no individual aroma compounds with the targeted aroma attribute. Semi-preparative HPLC was used to obtain less complex fractions of the wine extracts. The fractions were eluted in water/ethanol and, therefore, could be smelled directly. Fractions with the targeted aroma character were further resolved by GC-preparative fraction collection (GC-PFC). Recombinational GC-PFC demonstrated the importance of the components within a 4 min preparative GC fraction to the 'overripe orange' aroma of typical Bordeaux dessert wine. In Viognier wine, monoterpenes linalool, α-terpineol and geraniol as well as benzaldehyde were found to be associated with the 'apricot' character. Thus, several wine aroma compounds interact for these specific aromas to be perceived. This sensory-led combination of separation techniques is a powerful tool for the identification of key compounds responsible for specific aromas across the wine and beverage industries.

Symposium Introduction: Recent Progress and Current Challenges in Wine Analytical Sciences.

Œnology in its scientific dimension makes it possible to bring together a large community of researchers from various fields of expertise to share their knowledge and experience around wine. Two international meetings, the 11th International Œnology Symposium and In Vino Analytica Scientia 2019, were exceptionally held in the context of a joint organization under a common title ŒnoIVAS 2019, in Bordeaux, France, in June 2019. The conferences were attended by 350 delegates, from 24 countries, who shared different aspects of wine and spirits research, with 7 invited lectures, 71 oral communications, and over 200 posters. This special issue is a collection of full papers from a selection of contributed oral presentations and posters presented at the conference.

Aromatic Potential of Bordeaux Grape Cultivars: Identification and Assays on 4-Oxononanoic Acid, a γ-Nonalactone Precursor.

γ-Nonalactone has been demonstrated to be a chemical marker of dried/cooked fruit nuances detected in must and wine, but little is known about its formation pathways. Therefore, on the basis of the literature, we hypothesized 4-oxononanoic acid as a potential precursor. Using dichloromethane extraction followed by gas chromatography coupled to negative chemical ionization mass spectrometry, this keto acid was identified and quantified in Merlot and Cabernet Sauvignon musts. Its concentration ranged from traces to 60 µg/L. The biotransformation of 4-oxononanoic acid into γ-nonalactone by Saccharomyces cerevisiae during alcoholic fermentation was demonstrated using labeled d6-4-oxononanoic acid. Additional experiments shed light on the 4-oxononanoic acid role as a γ-nonalactone precursor and revealed that this biotransformation was (R)-enantioselective. Sensory and distribution studies of the enantiomers revealed that the detection threshold of R and S forms were 66 and 35 µg/L and the average ratio of R/S in grape and wine was 94:6 and 65:35.

Exploration of space to achieve scientific breakthroughs.

Living organisms adapt to changing environments using their amazing flexibility to remodel themselves by a process called evolution. Environmental stress causes selective pressure and is associated with genetic and phenotypic shifts for better modifications, maintenance, and functioning of organismal systems. The natural evolution process can be used in complement to rational strain engineering for the development of desired traits or phenotypes as well as for the production of novel biomaterials through the imposition of one or more selective pressures. Space provides a unique environment of stressors (e.g., weightlessness and high radiation) that organisms have never experienced on Earth. Cells in the outer space reorganize and develop or activate a range of molecular responses that lead to changes in cellular properties. Exposure of cells to the outer space will lead to the development of novel variants more efficiently than on Earth. For instance, natural crop varieties can be generated with higher nutrition value, yield, and improved features, such as resistance against high and low temperatures, salt stress, and microbial and pest attacks. The review summarizes the literature on the parameters of outer space that affect the growth and behavior of cells and organisms as well as complex colloidal systems. We illustrate an understanding of gravity-related basic biological mechanisms and enlighten the possibility to explore the outer space environment for application-oriented aspects. This will stimulate biological research in the pursuit of innovative approaches for the future of agriculture and health on Earth.

Contribution of Volatile Odorous Terpenoid Compounds to Aged Cognac Spirits Aroma in a Context of Multicomponent Odor Mixtures.

Cognac spirit aromas result from the presence of a wide variety of volatile odorous compounds associated with the modalities of spirit distillation and aging. However, very few studies have been carried out on aging notes of Cognac spirits. An HPLC fractionation approach was used in order to evidence fractions of interest recalling the specific aromatic nuances of aged Cognac. Then, a GC-O/MS analysis of the selected fractions allowed one to detect odorous zones and identify several volatile compounds. Among them, various well-known volatile compounds representative of the terpenoid family were highlighted, such as geraniol, α-terpinene, nerol, α-terpineol, 1,8-cineole (eucalyptol) and, particularly, piperitone, santalol, and α-campholenal, which have not previously been cited in Cognac. These compounds were quantitated and their detection thresholds were determined. Geraniol, α-terpinene, α-terpineol, and 1,8-cineole concentrations increased while spirits were more aged, while nerol tends to decrease. A sensory contribution of terpenes was observed through perceptual synergic effects, along with ß-damascenone and whisky lactone.