Aromatic potential of botrytized white wine grapes: identification and quantification of new cysteine-S-conjugate flavor precursors.

Sweet wines made from botrytized grapes contain much higher concentrations of volatile thiols, especially 3-sulfanylhexan-1-ol (3SH), than dry white wines. Three new specific volatile thiols (3-sulfanylpentan-1-ol (3SP), 3-sulfanylheptan-1-ol (3SHp), and 2-methyl-3-sulfanylbutan-1-ol (2M3SB) were recently identified in Sauternes wines. Like most volatile thiols, these compounds were almost totally absent from must, mainly being formed during alcoholic fermentation. In this work, we describe the identification and quantification of three new cysteine-S-conjugate precursors in must made from Botrytis-infected grapes. S-3-(pentan-1-ol)-L-cysteine (P-3SP), S-3-(heptan-1-ol)-L-cysteine (P-3SHp), and S-3-(2-methylbutan-1-ol)-L-cysteine (P-2M3SB) were identified by direct GC-MS analysis of their derivative forms obtained by silylation of an enriched fraction, isolated from must by affinity chromatography. Concentrations were considerably higher when Botrytis cinerea had developed on the grapes. In botrytized must, the mean levels of P-3SP, P-3SHp, and P-2M3SB were in the vicinity of 700, 50, and 500 nM, respectively, whereas concentrations in healthy must ranged from 0 to 50 nM. This indicated that these three new sulfanyl alcohols, responsible for the characteristic aroma of botrytized wines, were formed by the yeast metabolism during alcoholic fermentation from the corresponding non-volatile cysteine-S-conjugate precursors. Moreover, these results highlighted the predominant role of botrytization in developing grape aroma potential.

Impact of oxygen dissolved at bottling and transmitted through closures on the composition and sensory properties of a Sauvignon Blanc wine during bottle storage.

This work outlines the results from an investigation to determine the effect of the oxygen dissolved at bottling and the specific oxygen barrier properties of commercially available closures on the composition, color and sensory properties of a Bordeaux Sauvignon Blanc wine during two years of storage. The importance of oxygen for wine development after bottling was also assessed using an airtight bottle ampule. Wines were assessed for the antioxidants (SO(2) and ascorbic acid), varietal thiols (4-mercapto-4-methylpentan-2-one, 3-mercaptohexan-1-ol), hydrogen sulfide and sotolon content, and color throughout 24 months of storage. In addition, the aroma and palate properties of wines were also assessed. The combination of oxygen dissolved at bottling and the oxygen transferred through closures has a significant effect on Sauvignon Blanc development after bottling. Wines highly exposed to oxygen at bottling and those sealed with a synthetic, Nomacorc classic closure, highly permeable to oxygen, were relatively oxidized in aroma, brown in color, and low in antioxidants and volatile compounds compared to wines sealed with other closures. Conversely, wines sealed under more airtight conditions, bottle ampule and screw cap Saran-tin, have the slowest rate of browning, and displayed the greatest contents of antioxidants and varietal thiols, but also high levels of H(2)S, which were responsible for the reduced dominating character found in these wines, while wines sealed with cork stoppers and screw cap Saranex presented negligible reduced and oxidized characters.

Fluoride release and recharge characteristics of denture base resins containing surface pre-reacted glass-ionomer filler.

The flexural strength, flexural modulus, and the amount of fluoride released from four experimental denture base resins containing 5, 10, 20 and 30 wt% surface pre-reacted glass-ionomer (S-PRG) filler added to the powder were evaluated. The mean flexural strength of the experimental resins, except the 30 wt%, and the flexural modulus of all the resins, complied with ISO 1567 requirements. In the 20 wt% resin, the amount of fluoride released in the initial phase was 1.88 microg/cm2/day, after which the level decreased. After recharging in a 9000 ppm fluoride solution for eight hours, the level of released fluoride increased markedly to 40.21 microg/cm2/16 hrs. Our results show that fluoride levels increased as a function of the S-PRG filler content. After the almost completely discharged resins were recharged, similar fluoride release occurred again. These results suggest that denture base resins containing S-PRG filler have great recharge and release capabilities which may assist in preventing root caries of abutment teeth.

Quantification and odor contribution of 2-furanmethanethiol in different types of fermented soybean paste miso.

2-Furanmethanethiol, a compound contained in many kinds of food, was identified for the first time in five types of miso fermented soybean paste (red salty rice miso, thin-colored salty rice miso, weak salty rice miso, barley miso, and soy miso) by specific extraction of volatile thiols using p-hydroxymercuribenzoate. In the triangle test with red salty rice miso, which included a higher concentration of 2-furanmethanethiol, and thin-colored salty rice miso, which included a lower concentration of 2-furanmethanethiol, it was shown that the aroma of thin-colored salty rice miso was similar to that of red salty rice miso by adding 2-furanmethanethiol into thin-colored salty rice miso. In addition, a quantitative descriptive analysis (QDA) clearly shows that 2-furanmethanethiol contributed to the intensity of three odor qualities "thick, complex", "sweet", and "pleasant aroma like coffee beans" in six odor qualities and was a very important component of miso aroma.

Identification and characteristics of new volatile thiols derived from the hop (Humulus luplus L.) cultivar Nelson Sauvin (dagger).

Nelson Sauvin (NS) is a unique hop cultivar that was bred and grown in New Zealand. This hop gives a specific flavor (exotic fruit-like, white wine-like) to finished beers. However, the key compounds of this flavor have not yet been identified. We have attempted to identify the specific flavor compounds derived from NS. We focused on certain volatile thiols that are well-known to contribute to wine flavors, especially Sauvignon Blanc. The product made from NS (NS product) lost its specific flavor by contact with copper. Copper is well-known as an absorber of thiols in the field of wine flavor investigations. Therefore, it might point to the existence of thiols. We analyzed the NS product by GC-FPD, GC-olfactometry and GC-MS, and identified two new volatile thiols, 3-sulfanyl-4-methylpentan-1-ol (3S4MP), and 3-sulfanyl-4-methylpentyl acetate (3S4MPA). These compounds have a grapefruit-like and/or rhubarb-like odor, similar to that of Sauvignon Blanc. We quantified these compounds in the NS products and determined their thresholds. As a result, 3S4MP contained about 2-fold of its threshold in beers, and 3S4MPA was included below its threshold. However, it was confirmed that 3S4MP enhanced the flavors of 3S4MPA by synergy. Therefore, we concluded that both of the new volatile thiols would contribute to the specific odor of beers produced with NS.

Surprising structural lability of a cysteine-S-conjugate precursor of 4-methyl-4-sulfanylpentan-2-one, a varietal aroma in wine of Vitis vinifera L. cv. Sauvignon blanc.

4-Methyl-4-sulfanylpentan-2-one (1; 4MSP) provides a characteristic aroma compound of wines made from Vitis vinifera L. cv. Sauvignon blanc. 4MSP has a strong box-tree odor with a very low perception threshold and is derived from the cysteinylated precursor S-(1,1-dimethyl-3-oxobutyl)cysteine (4; P-4MSP). P-4MSP is transformed into 4MSP during alcoholic fermentation and is an excellent marker of varietal aroma potential. An improved synthesis of P-4MSP as well as of its deuterium-labeled analogue [D(6)]-P-4MSP is described. Several analytical methods (NMR, IR, LSI-MS, GC/MS, ESI-MS(n)) were combined to elucidate spontaneous reversible structural changes of P-4MSP at different pH values. At low pH, P-4MSP has a linear keto form. The keto-enol tautomerism was observed at neutral pH. At pH 8, the formation of N-substituted intramolecular hemiaminal was characterized by ESI-MS and ESI-MS(n) experiments. The hemiaminal loses H(2)O at high pH to produce a cycloimine, which is easily opened by acid hydrolysis. The keto-enol tautomerism explained the incorporation of only six D-atoms during the preparation of the P-4MSP deuterated standard even if [D(10)]mesityl oxide was used. Derivatization conditions for GC/MS analysis strongly affected the ratio of the monosilylated intramolecular cyclic form and the disilylated linear form of P-4MSP. The structural changes of P-4MSP may have a considerable impact on the development of methods of measuring varietal aroma potential.

Nitrogen catabolic repression controls the release of volatile thiols by Saccharomyces cerevisiae during wine fermentation.

Volatile thiols such as 4-methyl-4-sulfanylpentan-2-one (4MSP) and 3-sulfanylhexan-1-ol (3SH) are aromatic molecules having an important organoleptic impact on white wines. These components are produced from inodorous nonvolatile cysteinylated precursors by Saccharomyces cerevisiae metabolic activity during alcoholic fermentation. Here we provide a new insight into the genetic determinism of the production of volatile thiols by yeast. Using a gene deletion approach, we investigated the role of three yeast beta-lyases and demonstrate that Irc7p, a putative cystathionine beta-lyase, is one of the main proteins catalyzing the 4MSP and 3SH release under enological conditions. Moreover, we demonstrate that Ure2p/Gln3p proteins mainly control the bioconversion of volatile thiols by the transcriptional regulation of the IRC7 gene through the general mechanism of nitrogen catabolic repression. Finally, our findings suggest that the enantiomer balance of 3SH may be modulated by activating specifically stereoselective enzymes such as Irc7p.

Analysis of the diastereoisomers of the cysteinylated aroma precursor of 3-sulfanylhexanol in Vitis vinifera grape must by gas chromatography coupled with ion trap tandem mass spectrometry.

The diastereoisomeric distribution of S-3-(hexan-1-ol)cysteine (P-3SH), the cysteinylated precursor of 3-sulfanylhexan-1-ol (3SH) in Vitis vinifera grape juice, was determined by a new method. This procedure is based on the purification of P-3SH in a small volume of must (500 microL) by affinity chromatography, followed by the separation of chiral molecules in derivative forms by gas chromatography coupled with ion trap tandem mass spectrometry (GC-MS/MS). The diastereoisomers were easily separated using heptafluorobutyric anhydride and heptafluorobutanol (HFBA and HFOH) as derivatization reagents. Method validation was conducted using samples of grape juice, synthetic must, fermenting must, and wine that were fortified with P-3SH at concentrations of 0.6 and 2.5 microM. The relative standard deviation (RSD) and limit of detection (LOD) of the GC-MS/MS method were 4.6% and 1.5 nM, respectively. P-3SH assays in Bordeaux white grape juice affected by Botrytis cinerea showed an unusually increased proportion of the RS form of the precursor (approximately RR:RS=30:70) as compared to a diastereoisomer ratio (in the vicinity of 50:50) in healthy grape juice.

Odorous impact of volatile thiols on the aroma of young botrytized sweet wines: identification and quantification of new sulfanyl alcohols.

Specific extraction of volatile thiols using sodium p-hydroxymercuribenzoate revealed the presence of three new sulfanylalcohols in wines made from Botrytis-infected grapes: 3-sulfanylpentan-1-ol (II), 3-sulfanylheptan-1-ol (III), and 2-methyl-3-sulfanylbutan-1-ol (IV). The first two have citrus aromas, whereas the third is reminiscent of raw onion. In addition, 2-methyl-3-sulfanylpentan-1-ol, which has a raw onion odor, was tentatively identified. Like 3-sulfanylhexan-1-ol (I), already reported in Sauternes wines, compounds II, III, and IV were absent from must. They were found in wine after alcoholic fermentation, and their concentrations were drastically higher when Botrytis cinerea had developed on the grapes. In the commercial botrytized wines analyzed, the mean levels of II, III, and IV were 209, 51, and 103 ng/L, respectively. Despite their low odor activity values, sensory tests showed additive effects among I, II, and III, thus confirming their olfactory impact on the overall aroma of botrytized wines.

Stereoisomeric distribution of 3-mercaptohexan-1-ol and 3-mercaptohexyl acetate in dry and sweet white wines made from Vitis vinifera (Var. Sauvignon Blanc and Semillon).

The enantiomeric distribution of 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA) in Vitis vinifera wines was determined by combining two techniques: specific purification of volatile thiols from the wines using p-hydroxymercuribenzoate and separation of the chiral molecules by gas-phase chromatography on a cyclodextrin capillary column. The R and S enantiomer ratios of these two thiols in dry white Sauvignon blanc and Semillon wines are approximately 30:70 for A3MH and 50:50 for 3MH. However, in sweet white wines made from grapes affected by "noble rot" due to the development of Botrytis cinerea on ripe grapes, the proportion of the R and S forms of 3MH is in the vicinity of 30:70. During alcoholic fermentation, a change in the ratio of the two enantiomers of 3MH in dry white wines was observed. At the beginning of fermentation (around density 1.08), the S form represented over 60%; then, at lower density, as fermentation proceeded, the enatiomeric ratio approached 50:50. The ratio of the two 3MHA enantiomers remained constant throughout fermentation. On the contrary, the distribution of the two 3MH enantiomers changed very little during fermentation of the botrytized sweet wines. The perception thresholds for the R and S forms of 3MH in hydroalcoholic model solution are similar (50 and 60 ng/L). These two enantiomers have quite different aromas: The R form is fruitier, with a zesty aroma reminiscent of grapefruit, while the S form smells more of passion fruit. The perception thresholds of the R and S enantiomers of 3MHA are slightly different (9 and 2.5 ng/L). The less odoriferous R form is reminiscent of passion fruit, while the S form has a more herbaceous odor of boxwood.

Influence of fermentation temperature on volatile thiols concentrations in Sauvignon blanc wines.

The effect of Saccharomyces cerevisiae strains on the amount of 4-mercapto-4-methylpentan-2-one, a major varietal aroma of Sauvignon blanc wines, was demonstrated by previous research work. However, the influence of different alcoholic fermentation parameters on the levels of volatile thiols (4-mercapto-4-methylpentan-2-one, 3-mercaptohexan-1-ol and 3-mercaptohexyl acetate) in wines has not yet been investigated. The impact of fermentation temperature on the final amount of volatiles thiols and on some other analytical parameters (ethanol, total acidity, residual sugars, volatile acidity) was determined in a model medium and in grape juice. Interaction between fermentation temperature and yeast strain was also tested. The fermentation temperature influenced the amount of volatile thiols irrespective of the yeast strain used. The final levels of 4MMP and 3MH in model medium and in wines were higher when the alcoholic fermentation is conducted at 20 degrees C than at 13 degrees C. The 3MHA, which was correlated with the amount of 3MH determined in wines, was also higher when the alcoholic fermentation was conducted at 20 degrees C. From a technological point of view, the choice of yeast strain and fermentation temperature has a decisive influence on the concentrations of the varietal aromas of Sauvignon blanc wines.

A novel method for quantification of 2-methyl-3-furanthiol and 2-furanmethanethiol in wines made from Vitis vinifera grape varieties.

A rapid, easy method has been developed for isolating and quantifying 2-methyl-3-furanthiol (2M3F) in wines. Until now, it was not possible to quantify this highly odoriferous compound, with a smell reminiscent of cooked meat, in wine. The original aspect of this method is the specific release of volatile thiols using a cysteamine solution applied in reverse flow to sample percolation on the basis of a p-hydroxymercuribenzoate (pHMB)-volatile thiol conjugate formed by the direct addition of pHMB to 50 mL of wine. Purification of volatile thiols in wines is much faster and easier than our previous method. This method may also be used to assay 2-furanmethanethiol in wine. This thiol's strong aroma of roasted coffee has been shown to contribute to the "roast coffee" aroma of certain wines. Assaying 2M3F by this method showed that it was present in the wines analyzed (red and white Bordeaux, Loire Valley Sauvignon blanc, white Burgundy, and Champagne) at concentrations up to 100 ng/L, i.e., significantly above the olfactory perception threshold for this compound in model dilute alcohol solution.

Contribution of benzenemethanethiol to smoky aroma of certain Vitis vinifera L. wines.

Benzenemethanethiol, a volatile thiol with a strong empyreumatic aroma reminiscent of smoke, has been identified in boxwood (Buxus sempervirens L.) as well as in red and white Vitis vinifera L. wines. The perception threshold in a model hydroalcoholic solution is approximately 0.3 ng/L. All of the wines analyzed for this study contained this compound in concentrations of several dozen nanograms per liter. The Chardonnay wines had 30-40 ng/L. Sensory discrimination between a wine containing 7 ng/L benzenemethanethiol and the same wine with an additional 4 ng/L is very significant; the difference in smell is described as "empyreumatic". This compound can therefore significantly contribute to the aroma of certain wines (Sauvignon Blanc, Semillon, Chardonnay, etc.) containing concentrations as high as 30-100 times higher than their perception threshold.

Role of certain volatile thiols in the bouquet of aged champagne wines.

A method for the specific extraction of volatile thiols by use of p-hydroxymercuribenzoate has made it possible to identify certain flavor-active volatile thiols in Champagne wines. Benzenemethanethiol, 2-furanmethanethiol, and ethyl 3-mercaptopropionate were present in these wines at concentrations considerably higher than their perception thresholds. Their concentrations increased gradually in proportion to the bottle aging period and sharply as a result of disgorging. The contribution of these volatile thiols to the empyreumatic nuances of the bouquet of aged Champagne wines was demonstrated for the first time.

Sulfur aroma precursor present in S-glutathione conjugate form: identification of S-3-(hexan-1-ol)-glutathione in must from Vitis vinifera L. cv. Sauvignon blanc.

When Sauvignon blanc or Gros Manseng grape must was percolated through an immobilized gamma-glutamyltranspeptidase column, there was a significant increase in the concentration of S-3-(hexan-1-ol)-L-cysteine, the precursor of 3-mercaptohexan-1-ol, a compound that contributes to the varietal aroma of wines made from these grapes. Low- and high-resolution liquid secondary ion mass sepectrometry (LSIMS) analyses established the presence of S-3-(hexan-1-ol)-glutathione in Sauvignon blanc must. The identification of this compound suggests that the S-3-(hexan-1-ol)-L-cysteine in grapes is produced by the catabolism of S-3-(hexan-1-ol)-glutathione. As is the case in other plant or animal organisms, S-glutathione conjugates may be involved in certain detoxification systems in vines.