The effects of light exposure on 3-isobutyl-2-methoxypyrazine (IBMP) accumulation and degradation in Vitis vinifera L. cv. Cabernet Franc berries were assessed by comparison of shaded and exposed clusters within the same vine throughout a growing season. Twenty-seven vines were shoot-thinned to create regions of high and low cluster-light exposure within each vine. Samples were collected at 10 time points starting from 5 to 130 days postbloom. The experimental design allowed for intravine comparison of IBMP levels between treatments at each time. Vine-to-vine variability of IBMP and the correlation of IBMP to malic acid were also evaluated. Cluster exposure reduced accumulation of IBMP at all preveraison time points by 21-44%, but did not increase postveraison degradation. Significant vine-to-vine variability in IBMP content was observed, with the highest level of IBMP in shaded berries in the most vigorous block of vines. Although IBMP concentration by weight decreased significantly due to dilution just prior to color change (veraison), no significant IBMP degradation per berry occurred until after color change (day 70 postbloom). By contrast, malic acid degradation began prior to color change, and malic acid concentrations were not affected by cluster exposure preveraison, but were affected postveraison. A survey of 13 sites in New York state (Seneca Lake) showed that IBMP concentrations at 2 weeks preveraison were highly correlated (R(2) = 0.936, p < 0.0001) to levels at harvest, whereas classic grape maturity indices at harvest were uncorrelated with IBMP at harvest. In summary, light exposure conditions critically influence IBMP accumulation but not IBMP degradation.
Sunlight exposure of winegrape clusters is frequently reported to increase C(13)-norisoprenoids in resulting wines, but the timing and mechanism of this influence is not well understood. Fruit zone leaf removal was applied to Vitis vinifera cv. Riesling at three timings: 2, 33 and 68 days past berry set (PBS), and compared to an untreated control. Free and total 1,1,6-trimethyl-1,2-dihydronaphthalene (TDN), vitispirane and beta-damascenone were measured in juice and wines, and carotenoid profiles were determined in grapes at midseason and maturity. Significantly higher total TDN was observed in grapes from the 33-day PBS treatment compared to the control and other treatments (195 microg/L vs 54-87 microg/L). Total vitispirane in juice was also significantly increased in the 33-day PBS treatment, while total beta-damascenone was reduced in the 68-day PBS treatment compared to the control. Existing HPLC protocols were modified to allow for quantification of zeaxanthin in V. vinifera berries, and zeaxanthin was determined to be significantly higher in the 33-day PBS treatment than the control or other treatments (p < 0.05). Total TDN in juice correlated with free TDN in wine, with 11.0% +/- 2.5% of total juice TDN converted to free TDN in wine. In contrast, total vitispirane increased significantly during fermentation, and was not correlated with vitispirane in juice. In summary, leaf removal at 33 days PBS significantly increased zeaxanthin in Riesling grapes midseason, total TDN and vitispirane in the juice of mature Riesling grapes, and free and total TDN in finished wine, while earlier or later leaf removal had no effect.
Environmental factors affecting degradation of 3-isobutyl-2-methoxypyrazine (IBMP, "green pepper aroma") in wine grapes (V. vinifera) are widely studied, but the degradation pathway is not defined. We hypothesized that IBMP is demethylated to 3-isobutyl-2-hydroxypyrazine (IBHP) during fruit maturation effectively reversing the final putative step of IBMP biosynthesis. A quantification method for IBHP was developed using solid-phase extraction coupled to one- or two-dimensional gas chromatography-mass spectrometry with a recovery of ca. 80%. IBMP and IBHP in bell peppers (Capsicum annuum) and V. vinifera (cv. 'Cabernet Franc', 'Riesling', 'Pinot noir') were then measured at different maturities. IBMP and IBHP were inversely correlated in both bell peppers (R2=0.958) and Cabernet Franc grapes (R2=0.998) over a range of maturities. In bell peppers, we observed a significant decline in IBMP (125 to 15 ng/mL) and increase in IBHP (undetectable to 42 ng/mL) during ripening. In grapes, all cultivars had comparable IBHP concentrations preveraison (64 to 88 pg/mL) but differed in IBHP concentration by 2 orders of magnitude at the final sampling point (undetectable to 235 pg/mL). Higher preveraison IBMP was correlated with higher final IBHP across the three grape cultivars, with the order Cabernet Franc>Riesling>Pinot noir for both IBMP and IBHP. Acid hydrolysis resulted in a significant increase (33%) in IBHP in Cabernet Franc, indicating that IBHP exists partially in a bound form in grapes.
Although they possess significant viticultural advantages, interspecific hybrid grapes (Vitis spp.) are reported to produce wine with lower tannin concentrations than European wine varieties (Vitis vinifera). However, extensive quantitative data on this phenomenon as well as mechanistic explanations for these differences are lacking. A survey of primarily commercial wines from the Finger Lakes American Viticultural Area (New York) using a protein precipitation method determined that hybrid-based wines had >4-fold lower tannin concentrations than vinifera wines. To elucidate factors responsible for differences in wine tannin, 24 wines were produced from both red hybrid and vinifera cultivars under identical conditions. Lower wine tannin in French-American hybrid- than vinifera-based wines could be partially explained by lower grape tannin. However, experiments in which cell wall material was incubated with tannin indicated that cell wall binding may be of equal or greater importance in explaining lower wine tannin concentrations in hybrid-based wines. Subsequent characterization of cell wall material revealed that protein in flesh cell walls and, to a lesser extent, pectin in skin cell walls were correlated with cell wall binding.
Exogenous additions of condensed tannin (CT) to must or wine are a common winemaking practice, but many studies have reported inexplicably low and variable retention of added CT. We observed that additions of purified CT to red wines can result in the formation of an insoluble precipitate with high nitrogen content. Proteomic analysis of the precipitant identified several classes of pathogenesis-related proteins. Proteins in juices and red wines were quantitated by SDS-PAGE and were highest in native Vitis spp., followed by interspecific hybrids and Vitis vinifera. Wine protein was positively correlated with the ratio of juice protein to the quantity of tannin derived from fruit. The binding of added CT by wine protein could be well modeled by the Freundlich equation. These observations may explain the poor CT retention in previous studies, particularly for interspecific hybrids, and also indicate that protein removal during winemaking may improve exogenous CT retention.
Quantitation of trace-level (μg/L to ng/L) volatile compounds is routinely performed in a broad range of applications, including analyses of odorants, pesticide residues, or toxins in foodstuffs and related matrices. Conventional analyses based on gas chromatography-mass spectrometry (GC-MS) are limited by low throughput, and ambient approaches to sample introduction have typically had poor sensitivity. We prepared polydimethylsiloxane-coated stainless steel meshes for extraction and preconcentration of volatiles (Solid Phase Mesh Enhanced Sorption from Headspace, SPMESH), which could then be analyzed by Direct Analysis in Real Time (DART)-MS. The SPMESH cards were characterized by electron microscopy, and figures of merit for the approach were determined using two representative volatiles: 2-isobutyl-3-methoxypyrazine (IBMP) and linalool. Using DART-MS/MS and isotopically labeled internal standards, we achieved detection limits of 21 ng/L and 71 μg/L for IBMP and linalool in water. Good accuracy and precision could also be achieved for IBMP spikes in grape macerate, although accuracy for linalool was compromised by the presence of interferences. Detection limits could be further improved by an order of magnitude through the use of high resolution (HR) MS. Because extraction can be performed inexpensively in parallel and because it requires short data acquisition times (<1 min), SPMESH-DART-MS may be appropriate for high throughput trace level volatile analyses.
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