Direct HPLC analysis of quercetin and trans-resveratrol in red wine,grape, and winemaking byproducts

A simple and fast reversed-phase HPLC method using diode array detection was developed and validated for the simultaneous determination of trans-resveratrol and quercetin in Sicilian red wine from the Nero d'Avola red grape variety. Investigation was also extended to the quantitative determination of resveratrol and quercetin in grape skins and winemaking byproducts obtained from the same cultivar. Samples were eluted using a C18 narrow-bore column under isocratic conditions in less than 20 min. Quantification of trans-resveratrol and quercetin in red wine was performed without any sample pretreatment, whereas the determination of these phenolic compounds in grape skins and wine pomage required a solvent extraction procedure. Linearity was demonstrated over the 0.39-12.5 and 0.45-57.6 microg/mL range for trans-resveratrol and quercetin, respectively. Detection limits in real samples were in the low ppm level (0.07 mg/L for trans-resveratrol and 0.12 mg/L for quercetin). The HPLC-UV/DAD method was applied for the routine analyses of red wine and grape skin and winemaking byproduct extracts to evaluate their trans-resveratrol and quercetin content. In particular, a very high content of quercetin was found in wine pomace, suggesting the use of this wine byproduct as a potential source of this health-promoting phenolic compound.     

Determination of the grape invertase content (using PTA-ELISA) following various fining treatments versus changes in the total protein content of wine. relationships with wine foamability

Proteins have proven to play a major role in the stabilization of foam in Champagne wines despite their low concentration that ranges from 4 to 20 mg/L. The aim of this study was to evaluate the effect of fining on total protein and grape invertase contents of champenois base wines and their foaming properties. Data showed that fining and especially the use of bentonite at doses ranging from 10 to 50 g/hL leads to a significant decrease in the total protein content of wines together with that of the grape invertase content, with such a decrease being very detrimental to the foaming properties of the treated wines in terms of foam height (HM) and foam stability (HS). Only a slight decrease in the total protein content, in the grape invertase concentration, and in the foam quality of wines was observed when using casein (10 and 20 g/hL) or bentonite combined with casein (both at 20 g/hL). Our study thus clearly establishes the good correlation existing between the wine protein concentration and its foaming properties. A remarkable correlation was observed between the decrease in the grape invertase content and the total protein content of wines, following bentonite treatments, suggesting that the grape invertase (which represents at least 10-20% of the wine proteins) follows a similar behavior upon fining to other proteins of Champagne wines, despite the high molecular mass and the highly glycosylated structure of this particular protein. Moreover, the decrease in total protein and grape invertase contents of wine after fining with bentonite was found to be correlated with a decrease in the foaming properties of the corresponding wines (with respectively R(2) = 0.89 and 0.95).     

Determination of rotundone, the pepper aroma impact compound, in grapes and wine

Shiraz, also known as Syrah or Hermitage, is one of Australia's most popular red wine varieties both domestically and internationally. Black pepper aroma and flavor are important to some Australian Shiraz red wine styles. Recently, rotundone (a bicyclic sesquiterpene) was identified as the potent aroma compound responsible for pepper aromas in grapes, wine, herbs, and spices, including peppercorns. Here the development, optimization, and validation of the analytical method for the quantitative determination of rotundone in grapes and wine are described and discussed. The method is precise, accurate, robust, and sensitive with a subpart per trillion limit of quantitation. The method uses stable isotope dilution analysis with d(5)-rotundone as internal standard, solid-phase extraction and microextraction, and gas chromatography-mass spectrometry.     

Aging effects and grape variety dependence on the content of sulfur volatiles in wine

Thirteen sulfur compounds (boiling points from 35 to 231 degrees C), usually considered as possible off-flavoring volatiles, were quantified by a concurrent headspace-solid phase microextraction method coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS) on 80 not off-flavoring wines of four varieties (Merlot, Marzemino, and Teroldego as red wines and Chardonnay as a white one) and of five vintages produced in the North Italian Trentino region. The results of the research, the first Italian data-bank per variety on such volatiles, allow us to make a comparison with the data of other winegrowing areas, to investigate the aging effect on the considered volatiles, and, finally, to try a variety discrimination using statistical procedures. Dimethyl sulfide, 3-(methylthio)-1-propanol, diethyl sulfide, and diethyl disulfide were found to increase with time whereas 2-mercaptoethanol and ethylmercaptan showed a decreasing trend. Furthermore, the concentration of several compounds was found to be dependent on the variety. For instance, sulfide, disulfides, benzothiazole, and thioalcohols are at higher levels in Merlot wines, whereas thiols and thioacetates are more abundant in Marzemino and Teroldego wines. Chardonnay products, well apart from the other wines, are the poorest in 3-(methylthio)-1-propanol and rather rich in dimethyl disulfide and in diethyl disulfide, mostly in the aged wines. Applying the principal component analysis to the data, it was possible to demonstrate that Chardonnay and Merlot wines are well-discriminated from the Italian native varietal wines, which on their turn are only partially distinguishable among them. A contribution of these compounds to the variety characteristics of wine is reasonable.     

Liquid chromatography-electrospray tandem mass spectrometry of cis-resveratrol and trans-resveratrol: development, validation, and application of the method to red wine, grape, and winemaking byproducts

The application of liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) was investigated for the analysis of trans-resveratrol in red wine, grape skin, grape pomace, and winemaking byproducts. Chromatographic elution performed under isocratic reversed-phase conditions using a C18 narrow-bore LC column allowed retention times lower than 12 min to be obtained. Qualitative analysis was performed in negative-ion (NI) full-scan and product-ion scan acquisition modes, whereas method validation in terms of linearity, detection limits, accuracy, and robustness was carried out under NI selected reaction monitoring conditions. The matrix-matched detection limit was calculated in the low parts per billion range (10 microg/L). Results of the application of the method to red wine, grape, and winemaking byproduct samples were compared with those obtained using an LC-UV/DAD technique. Determination of trans-resveratrol in the samples investigated showed that the highest concentration was found in red wine, whereas wine made from grape pomace contained the lowest content.     

霞多丽干白葡萄酒品种香和发酵香成分变化的比较研究

本研究对确立宁夏贺兰山东麓原产地葡萄酒质量的感官体系标准,以及对葡萄酒生产工艺优化和质量评价有着重要实用价值。采用溶液萃取法提取霞多丽品种果实与干白发酵香气成分,用气相色谱-质谱进行分离测定,结合计算机检索技术对分离化合物进行鉴定,应用色谱峰面积归一法测定各成分的相对含量。实验结果：果实和干白葡萄酒中分别分离出81和33个峰,鉴定出78和32个香气化学成分,共占其色谱流出组分总量的98.89%和98.97%;成熟果实的挥发性物质,主要是2-呋喃甲醛等脂肪醛、2-呋喃乙酮等脂肪酮和十六碳酸等脂肪羧酸类。经过发酵工艺处理原酒中挥发性物质,主要有3-甲基丁醇、丁二酸二乙酯、辛酸、苯乙醇等。相对含量较高的香气成分种类相似,而微量特征香气成分差异较为显著,从而构成不同产区同一品种葡萄酒相似和独特香气与风格。本结果将对不同原产区优质葡萄酒酿造工艺生产,最大限度的释放游离态芳香物质的能力,提高干白葡萄酒的果香,有重要指导意义。     

葡萄与葡萄酒中的单宁及其与葡萄酒的关系

单宁是葡萄和葡萄酒中重要的组成成分.本文就葡萄与葡萄酒中单宁的分类、组成、及其对葡萄酒品质的影响及单宁与人体健康等几方面进行了综述.并对今后研究重点提出展望.     

Influence of grape density and harvest date on changes in phenolic composition, phenol extractability indices, and instrumental texture properties during ripening

Changes in the phenolic composition, phenol extractability indices, and mechanical properties occur in grape berries during the ripening process, but the heterogeneity of the grapes harvested at different ripening stages affects the reliability of the results obtained. In this work, these changes were studied in Nebbiolo grapes harvested during five consecutive weeks and then separated according to three density classes. The changes observed in chemical and mechanical parameters through the ripening process are more related to berry density than harvest date. Therefore, the winemaker has to select the flotation density according to the objective quality properties of the wine to be elaborated. On the other hand, the stiffer grapes were associated with a higher accumulation of proanthocyanidins. The harder grapes provided the higher concentration and extractability of flavanols reactive to vanillin, whereas the thicker ones facilitated the extraction of proanthocyanidins.     

Fate of pesticides during the winemaking process in relation to malolactic fermentation

The effect of red wine malolactic fermentation on the fate of seven fungicides (carbendazim, chlorothalonil, fenarimol, metalaxyl, oxadixyl, procymidone, and triadimenol) and three insecticides (carbaryl, chlorpyrifos, and dicofol) was investigated. After malolactic fermentation using Oenococcus oeni, which simulated common Australian enological conditions, the concentrations of the active compounds chlorpyrifos and dicofol were the most significantly reduced, whereas the concentrations of chlorothalonil and procymidone diminished only slightly. The effect of these pesticides on the activity of the bacteria was also studied. Dicofol had a major inhibitory effect on the catabolism of malic acid, whereas chlorothalonil, chlorpyrifos, and fenarimol had only a minor effect.     

不同施锌量对酿酒葡萄“赤霞珠”锌含量及其品质的影响

以4年生"赤霞珠(Cabernet Sauvignon)"葡萄为研究对象,进行连续2年滴灌[0 kg/hm~2(CK)、24.5 kg/hm~2(T_1)、34.0 kg/hm~2(T_2)、43.5 kg/hm~2(T_3)、53.0 kg/hm~2(T_4)、62.5 kg/hm~2(T_5)]和转色期叶面喷施[0 kg/hm~2(CK)、0.5 kg/hm~2(T_1)、1.0 kg/hm~2(T_2)、1.5 kg/hm~2(T_3)、2.0 kg/hm~2(T_4)、2.5 kg/hm~2(T_5)]两种方式配合施EDTA-Zn的田间试验,探讨强碱性灰钙土(pH8.5)补充锌肥对酿酒葡萄锌含量、生长和品质的影响。结果表明:与不施锌肥(CK)相比,施锌肥处理酿酒葡萄锌含量、粒径、穗长、百粒重和产量分别增加了42.1%～78.7%、2.6%～28.2%、10.1%～22.3%、3.0%～8.8%和17.2%～216.6%。同时,与不施锌(CK)相比,酿酒葡萄的单宁含量降低42.0%～67.9%,花色苷、总酚、可溶性固形物和可溶性糖含量分别提高了0.7%～71.9%、5.0%～80.6%、9.3%～24.0%和10.4%～29.4%,而可滴定酸含量除T_4外没有显著增加。相关分析表明,锌含量与穗长、百粒重、粒径、产量、总酚和可溶性糖均显著相关。综合酿酒葡萄锌含量和品质分析得出T_3处理效果最好。在宁夏贺兰山东麓石灰性土壤条件下,EDTA-Zn施用量45 kg/hm~2左右对于提高酿酒葡萄的锌含量、产量和品质较为适宜。     

葡萄果实和葡萄酒中缩合单宁的研究进展

缩合单宁是葡萄与葡萄酒中重要的组分之一,它影响着葡萄酒的色泽、风味、稳定性和贮藏寿命,与人的健康也有密切关系;该文介绍了缩合单宁的作用、结构及其与原花色素的关系,葡萄与葡萄酒中缩合单宁的存在形式及其在葡萄果实成熟和葡萄酒酿造过程中的变化和对葡萄酒口感的影响及缩合单宁的生物合成等方面的研究进展,提出了今后的研究方向.     

Analysis of grape and wine anthocyanins by HPLC-MS

The development and application of valuable analytical tools suitable for the varietal authentication of premium red wines are matters of interest in order to avoid fraud. In this study, an HPLC-MS procedure has been developed using trifluoroacetic acid as an acid modifier in the mobile phase. This method may be used as a routine method using UV-vis detection and allows the simultaneous analysis of the structural features of anthocyanins by MS under the same chromatographic conditions. Twenty different anthocyanins have been detected in 19 different samples of both grape extracts and wines. Cis and trans isomers of p-coumaryl derivatives have been identified for the first time. Important qualitative and quantitative differences among cultivars have been detected.     

转色期叶面氮素调控对酿酒葡萄'赤霞珠'品质的影响

宁夏贺兰山东麓现阶段水肥一体轻简化栽培模式下酿酒葡萄对氮素营养需求发生了巨大变化,如果继续沿用传统氮肥施用方式可能会引发酿酒葡萄成熟过快、浆果内含物累计不足、果皮中酵母可同化氮偏低等问题。以喷清水为对照,在酿酒葡萄转色期内叶面喷施硫酸铵、硝酸铵钙、尿素、苯丙氨酸及谷氨酸5种水溶性氮源,研究不同氮源对酿酒葡萄'赤霞珠'品质的影响。结果表明：转色期叶面氮素调控的葡萄浆果可溶性固形物以硫酸铵处理最佳,高达29.21%;花色苷在尿素处理下含量高达2.28 mg/g,较其余氮素处理提高16.33%～55.10%;单宁含量以苯丙氨酸处理最佳,高达19.88 mg/g;总酚含量在苯丙氨酸处理下高达19.56 mg/g,较其余氮素处理提高8.91%～27.34%。综合分析得出,转色期叶面喷施苯丙氨酸和尿素两种氮源均可改善酿酒葡萄生长发育,提升浆果品质。     

Evolution of volatile byproducts during wine fermentations using immobilized cells on grape skins

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae cells on grape skins. Repeated batch fermentations were conducted using this biocatalyst as well as free cells, at 25, 20, 15, and 10 degrees C. Solid phase microextraction (SPME) was used in monitoring the evolution of volatile byproducts. The effect of immobilization and temperature on evolution patterns of volatiles was obvious. The major part of esters was formed after consumption of 40-50% of the sugars. Similar processes were observed for amyl alcohols and 2-phenylethanol, whereas 1-propanol and 2-methyl-1-propanol were formed during the whole alcoholic fermentation period at an almost constant formation rate. Acetaldehyde and acetoin were synthesized in the early stages of fermentation. Afterward, their amount decreased. In most cases, immobilized cells exhibited higher formation rates of volatiles than free cells. The final concentration of esters was higher in wines produced by immobilized biocatalyst. Their amount increased with temperature decrease. The opposite was observed for higher alcohols.     

基于GIS的宁夏酿酒葡萄种植区划

根据宁夏1∶25万地理信息数据和宁夏各气象站1960年-2005年的气候资料，在运用GIS技术对宁夏气候要素网格推算的基础上，利用全国北方主要酿酒葡萄产区取样化验资料，将7～8月份水热系数作为一级区划指标，7～9月大于等于10℃积温作为二级区划指标，同时考虑了土壤类型和灌溉条件的影响，采用多边形叠置方法对宁夏酿酒葡萄种植区域进行气候精细区划，得到作物农业气候区划空间数据集合。对生成的各种资源空间数据 ,进行多种图形整饰和矢量化处理，制作出250 m空间分辨率的宁夏酿酒葡萄气候区划图和种植区划图。区划结果表明：宁夏开发优质酿酒葡萄基地的潜力很大，宁夏酿酒葡萄种植可分为4个区。酿酒葡萄种植特优区分布在包括贺兰山东麓在内的宁夏灌区周边风沙土区域，面积达1890 km²；优质区主要集中在宁夏平原，贺兰山东麓，清水河流域等，面积约为20580 km²，这一区域光热资源丰富、昼夜温差大、水热系数小于1.0，生长季降水量小于220 mm，有好的灌溉条件。适宜区包括中部干旱带的大部分地区和卫宁平原的中卫；宁南阴湿区光热不足，年积温少于2700℃，不宜种植酿酒葡萄。     

Improving grape phenolic content and wine chromatic characteristics through the use of two different elicitors: methyl jasmonate versus benzothiadiazole

Benzothiadiazole (BTH) and methyl jasmonate (MeJ) have been described as exogenous elicitors of some plant defense compounds, polyphenols among them. The objective of this study was to determine whether the application of BTH or MeJ to grape clusters at the beginning of the ripening process had any effect on the accumulation of the main flavonoid compounds in grapes (anthocyanins, flavonols, and flavanols) and the technological significance of these treatments in the resulting wines. The results obtained after a 2 year experiment indicated that both treatments increased the anthocyanin, flavonol, and proanthocyanidin content of grapes. The wines obtained from the treated grapes showed higher color intensity and total phenolic content than the wines made from control grapes. The exogenous application of these elicitors, as a complement to fungicide treatments, could be an interesting strategy for vine protection, increasing, at the same time, the phenolic content of the grapes and the resulting wines.     

Rapid method for proline determination in grape juice and wine

Proline is typically the most abundant amino acid present in grape juice and wine. The amount present is influenced by viticultural and winemaking factors and can be of diagnostic importance. A method for rapid routine quantitation of proline would therefore be of benefit for wine researchers and the industry in general. Colorimetric determination utilizing isatin as a derivatizing agent has previously been applied to plant extracts, biological fluids, and protein hydrolysates. In the current study, this method has been successfully adapted to grape juice and wine and proved to be sensitive to milligram per liter amounts of proline. At sugar concentrations above 60 g/L, interference from the isatin-proline reaction was observed, such that proline concentrations were considerably underestimated in grape juice and dessert wine. However, the method was robust for the analysis of fermentation samples and table wines. Results were within ±10% agreement with data generated from typical HPLC-based analyses. The isatin method is therefore considered suitable for the routine analysis required to support research into the utilization or release of proline by yeast during fermentation.     

Effect of oak extract application to Verdejo grapevines on grape and wine aroma

Volatile compounds from a commercial aqueous oak extract application to white Verdejo grapevines at veraison have been studied. Treated grapes under two types of formulation (25% and 100%) have been analyzed at the optimum maturation time, and winemaking was then subsequently carried out. The volatile compounds were analyzed by stir bar sorptive extraction-gas chromatography-mass spectrometry. The results suggest that after the grapevine treatments, grapes store the volatiles in the form of nonvolatile precursors, and some of the volatiles are released during the winemaking process, especially six months after the alcoholic fermentation. The sensory analysis shows that wines maintain the typical aroma properties of Verdejo wines at the end of fermentation; but after six months, the wine color is greener and more astringent, and, in terms of aroma, it has wooden notes as if the wine has been aged in oak barrels.     

Thaumatin-like proteins and chitinases, the haze-forming proteins of wine, accumulate during ripening of grape (Vitis vinifera) berries and drought stress does not affect the final levels per berry at maturity

Thaumatin-like proteins and chitinases, which are pathogenesis-related (PR) proteins, were the major soluble protein components of grapes from five cultivars of Vitis vinifera. This dominance of PR proteins was apparent at berry softening (véraison) and then throughout berry development for the Muscat of Alexandria, Sultana, and Shiraz cultivars and in the berries of the Sauvignon Blanc and Pinot Noir cultivars examined at commercial maturity. The M(r) of the major thaumatin-like protein from Muscat of Alexandria grapes was 21 272, and those of the three major chitinases from this cultivar, ChitB, ChitC, and ChitD, were 25 588, 25 410, and 25 457, respectively. The vines in the study were irrigated and showed no obvious signs of disease. Shiraz vines that had not been irrigated throughout the season were clearly water stressed, but had levels of PR proteins in the berry similar to vines that had been fully irrigated. It appears that the production of PR proteins that cause protein instability in wines by grapes may be little influenced by environmental conditions.