Simultaneous Quantification of Primary, Secondary Amino Acids, and Biogenic Amines in Musts and Wines Using OPA/3-MPA/FMOC-CI Fluorescent Derivatives

ABSTRACT: Free amino acids and biogenic amines occur in musts and wines and affect the course of fermentation in musts and the quality of wines. Each sample was analyzed in 138 min and each analysis involved quantification of 23 amino acids and 10 amines. The detection limit for proline was 27.3 mg/L. The mean detection limit for amino acids was 0.58 mg/L and for amines 0.28 mg/L. Precision was 0.5 to 19.2% for wine samples and accuracy by the standard addition method was 99.8% (recovery value). In all, 33 wines and 39 must samples were analyzed. In wines, ethanolamine, histamine, ethylamine, and putrescine amounted to 93% of the amine content.     

Varietal and geographic classification of French red wines in terms of elements,amino acids and aromatic alcohols

Thirty-four French red wines from three regions already studied for their anthocyanin and flavonoid constituents have been further analysed for elements, amino acids and aromatic alcohols. An interpretation of the differences between wines related to their different geographic and varietal origins has been made from the results of statistical analyses: F statistic, principal component analysis (PCA) and stepwise discriminant analysis (SDA). Wine samples produced near Bordeaux were found to be characterised by higher rubidium and lower lithium and calcium concentrations. Differences between wine samples made from the same grape variety or produced in the same region are mainly related to differences in potassium and ash contents. The PCA demonstrates that the concentration of most amino acids in wines is mainly a function of the technology used in wine production. Moreover, it shows that the concentration of proline, hydroxyproline and ethanolamine are generally lower in wines prepared from Carignan, Cinsault and Grenache grapes than in wines from Cabernet Franc, Cabernet Sauvignon or Merlot grapes. Finally, it shows clear differences between wines according to the latitude of the production centre. These differences correspond to increasing total nitrogen content from south to north, ie Narbonne and Bordeaux versus Angers. In discriminant analysis Bordeaux wines were characterised by high rubidium and low arginine concentrations, Angers wines by a lower arginine content and Narbonne wines by their abundance of magnesium and deficiency of ethanolamine.     

Comparison between the phenolic composition of Petit Verdot wines elaborated at different maceration/fermentation temperatures

The objective of this work was to contribute to the knowledge of the relationship between phenolic composition and the temperature of maceration/fermentation on Petit Verdot red wines, which were made with three treatments (17°C, 21°C, and 25°C). The phenolic compounds of Vitis vinifera cv. Petit Verdot wines elaborated at different maceration/fermentation temperatures were determined and compared by high-performance liquid chromatography coupled with diode array detector and electrospray ionization mass spectrometry. A total of 45 phenolic compounds were detected in all wines. The phenolic composition was affected by the temperature of maceration. The increment of maceration/fermentation temperature had a positive effect on the total concentration of the phenolic compounds and chromatic characteristics of these wines. The colour of Petit Verdot young red wines showed more colour intensity and their luminosity descended due to the increase in temperature. The highest total content of anthocyanins was determined in wines macerated at 21°C. However, other groups of phenolic compounds (flavonols, proanthocyanidins, and stilbenes) increased their total content by raising the maceration/fermentation temperature, reaching maximum values at 25°C.     

Authentication of Italian red wines on the basis of the polyphenols and biogenic amines

Seventy-three Southern Italian red wines were characterized according to their content in total polyphenols, trans- and cis-resveratrol and biogenic amines. These quality parameters were used in multivariate statistical analysis to discriminate the wines according to their specific geographical origin. The results indicated that total polyphenols, resveratrol isomers and biogenic amines provide a good prospect for discriminating wines by regions. The discrimination was also possible to a lesser extent by cultivar. In particular, canonical variate analysis suggested that the discrimination of wines according to their provenance is based on the following parameters: cis-resveratrol, total polyphenols, spermidine and tryptamine for Basilicata region; agmatine and trans-resveratrol for Calabria and Campania regions; cadaverine, ethanolamine, histamine, putrescine and tyramine for Puglia region.     

Determination of ochratoxin A in wine by immunoaffinity cleanup and liquid chromatography tandem mass spectrometry

A simple and accurate method has been developed for determining ochratoxin A (OTA), using an immunoaffinity column for cleanup and liquid chromatography-tandem mass spectrometry for identification and quantification. Wine samples were diluted with a solution containing polyethylene glycol 8000 and sodium hydrogen carbonate, filtered through a glass microfiber filter, and cleaned up on an immunoaffinity column. OTA was then eluted with methanol-acetic acid (98:2) and analyzed by liquid chromatography-tandem mass spectrometry. The average recoveries of OTA from red and white wines were 95 and 96.7% (spiked OTA level was 0.05 ng/ml) and repeatabilities (relative standard deviation) were 3.8 and 2.4%, respectively. The detection limit was 0.0003 ng/ml based on the signal-to-noise ratio in wine of 3:1. Analysis of 74 samples of domestic and imported wines showed OTA levels ranging from < 0.0003 to 0.82 ng/ml, with an incidence of contamination of 92.1% for red wines, and < 0.0003 to 0.51 ng/ml, with an incidence of contamination of 77.8% for white wines. These detection rates were higher than those rates of past reports of OTA contamination in wine, due to the high sensitivity of this method. However, all samples analyzed in this study complied with European Union regulations. It is concluded that this method is a useful tool for the quality assurance of wine.     

超高效液相色谱串联质谱法测定樱桃酒中的5种酚酸

建立了超高液相色谱-串联四级杆质谱联用(UPLC-MS/MS)技术同时检测樱桃酒中没食子酸、4-羟基苯甲酸、绿原酸、香草酸和咖啡酸5种生物活性酚酸的方法。采用Waters Acquity UPLC BEH色谱柱(100×2.1mm,1.7μm),流动相为甲醇-0.1%甲酸,在梯度条件下进行分析,分析物在负离子模式下用保留时间和质谱信息进行定性,峰面积进行定量,并评价其定量分析的精密度、线性和检出限。结果显示,回收率在82.3%～110.8%之间;相关系数r>0.99,线性关系良好;精密度的相对标准偏差RSD<2.0%,检出限为0.020～0.050mg/L,说明该方法简便、快速、准确、灵敏,可用于樱桃酒中酚酸的检测。此外,实验采用SPSS17.0软件对樱桃酒的理化指标和5种酚酸进行了简单相关分析。     

Free amino acids and biogenic amines in Alicante Monastrell wines

The simultaneous determination of 17 free amino acids and 8 biogenic amines in Alicante Monastrell wines was investigated for the first time. The quantification was carried out by using a RP-HPLC method, based on a pre-column derivatization with o-phthaldialdehyde (OPA) and fluorescence detection. From the results obtained it may be concluded that the most abundant free amino acids were Glu, Arg, Ala Asp, and Lys. None of the wine samples analysed had histamine (HIM) or Tyramine (TYM) levels above the limits considered as a possible toxic risk for healthy individuals. No measurable amounts of cadaverine (CAD) or methylamine (MEA) were found, showing no spoilage symptoms of sensory properties of the wines. Tryptamine (TRM) content was significantly higher in aged wines compared to young wines. However ethanolamine (ETA) content was lower. These data were used to make a preliminary classification of the samples using cluster analysis.     

A method for the determination of methyl carbamate and ethyl carbamate in wines.

A method is described for the simultaneous determination of methyl carbamate (MC) and ethyl carbamate (EC) in wines that is based on: (a) extraction of the sample with dichloromethane using an extraction tube or an alumina-Celite column, (b) concentration of the extract to a small volume, and (c) determination by gas-liquid chromatography-thermal energy analyser (N-mode). The method is highly sensitive (1-2 ng/ml), accurate (recoveries greater than 80%), and precise (CV, 5-10%). Nineteen of 27 samples of wines analysed contained traces (up to 2.7 ng/ml) of MC, and most contained EC (up to 70 ng/ml). Wines treated in the laboratory with 200 ppm dimethyl pyrocarbonate (DMPC)-a cold sterilant recently approved for use in wines-indicated that such a treatment may increase the MC contents of the wines to 10 ng/ml. Additional studies suggested that formation of MC in DMPC-treated wines is dependent on both pH and ammonia content of the wines. The identity of MC in a few selected samples was confirmed by gas-liquid chromatography-high resolution (10 K) mass spectrometry. The natural low levels of MC found in these wines are not considered to pose a risk to human health.     

Antioxidant activities of volatile and non‐volatile fractions of selected traditionally brewed Korean rice wines

Antioxidant activities of traditionally brewed Korean rice wines have recently been reported. The objective of this study was to identify the antioxidant activities of the volatile and non‐volatile fractions of three Korean rice wines, and further to identify the active ingredients responsible for the antioxidant properties. Volatile and non‐volatile components of three rice wines, Measilju, Kukwhaju and Gugijaju, were separated into seven fractions according to their polarities. Antioxidant activities were determined by an aldehyde–carboxylic acid assay for the volatile fractions and a lipid–malondialdehyde assay for the non‐volatile fractions. Volatile constituents of each fraction were identified and quantified using gas chromatography–mass spectrometry. In addition, antioxidant activities of specific authentic chemicals from the volatile fractions of the wines were measured, and the antioxidant properties of the volatile fractions were determined. Identification of volatile compounds potentially showing high antioxidant activities were conducted in each fraction of the three wines. Among the volatile compounds identified from volatile fractions of the wines, benzeneethanol, 2‐furanmethanol and 4‐ethyl‐2methoxyphenol showed the highest antioxidant activities, when added at higher concentrations (~100 µg/g). Identification of antioxidant properties, as well as the specific compounds responsible for these antioxidant properties, could aid Korean rice wine product developers, in terms of strategic planning for sales in the Korean market. Copyright © 2014 The Institute of Brewing & Distilling     

The multi-element determination and regional discrimination of Australian wines

Concentrations were determined for 56 elements in 1397 samples of Australian wine, using solution based inductively coupled plasma mass spectrometry and emission spectroscopy. Interpretation of the data indicated that, within red and within white wines, the vintage and grape variety had little effect on the multi-element composition of the wine, although significant differences were observed between red and white wines.     

Volatile Composition of Macedonian and Hungarian Wines Assessed by GC/MS

The volatile composition of eight varietal wines Vranec, Merlot, Cabernet Sauvignon, Temjanika and Chardonnay from the Republic of Macedonia, and Portugieser, Kékfrankos and Tokaji Aszú from Hungary has been characterized by means of gas chromatography/mass spectrometry technique. The wine volatile compounds were extracted in dichloromethane, and the extracts were concentrated under nitrogen. Forty-four volatile compounds have been identified mainly using the NIST mass spectral library and by comparison with the available standards used for quantification as well. Differences between the wines were noted for a number of compounds, such as a higher concentration of 1-pentanol and 2-phenyl ethanol in the red wines. Monoterpenes, linalool and terpineol were detected only in the white wines, Chardonnay and Tokaji. Macedonian red wines were characterized by a higher level of alcohols, while the Hungarian wines contained a higher amount of esters, fatty acids and lactones. A statistical treatment including one-way ANOVA, followed by a Tukey’s test has been performed in order to ascertain possible significant differences between the wines studied, and principal component analysis to study the possible grouping of the wines.     

Identification and analysis of 2-chloro-6-methylphenol, 2,6-dichlorophenol and indole: causes of taints and off-flavours in wines

Background and Aims: The aim of this study was to determine the cause of taints and off-flavours in a number of commercial wines and to develop methods for quantitative analysis of the compounds responsible. Methods and Results: Three compounds, 2-chloro-6-methylphenol (i.e. 6-chloro-ortho-cresol), 2,6-dichlorophenol and indole, were identified by gas chromatography (GC)/mass spectrometry (MS)/olfactometry as causes of taints or off-flavours in the wines. New methods for quantitative analysis of these three compounds, as well as some other chloro- and bromophenols were developed. The methods employed GC/MS and stable isotope dilution analysis, and confirmed the presence of 2-chloro-6-methylphenol, 2,6-dichlorophenol and indole in some wines at concentrations well above their odour and taste detection thresholds. Conclusions: 2-Chloro-6-methylphenol, 2,6-dichlorophenol and indole were confirmed as causes of taints and off-flavours in commercial wines. Significance of the Study: The development of new analytical methods will make it possible to determine whether chlorophenol (and perhaps also bromophenol) taint in wine is more widespread than previously recognised. Similarly, the simple, rapid and accurate method for quantifying indole in wine will allow large numbers of wine and ferment samples to be studied to determine those factors that can result in unacceptably high concentrations in commercial wines.     

龙眼干白葡萄酒发酵过程中生物胺和氨基酸含量的变化

本研究利用高效液相色谱技术分析了龙眼干白葡萄酒酒精发酵和苹乳发酵过程中8种生物胺和22种氨基酸含量的变化。结果表明,在酒精发酵过程中8种生物胺含量均很低,而在苹乳发酵过程中迅速增加。发酵结束时干白葡萄酒中亚精胺和乙醇胺含量最高,其次为组胺和羟色胺。这4种生物胺的前体氨基酸在苹乳发酵阶段含量没有明显变化,而其它大部分氨基酸在酒精发酵过程中含量下降,在苹乳发酵过程中含量呈现不同程度升高。因此,我们认为：葡萄酒发酵过程中氨基酸含量变化与其对应生物胺的变化之间没有相关性,葡萄酒中生物胺的组成比例可能主要取决于乳酸菌中氨基酸脱羧酶的种类和活性。     

Occurrence of ethyl carbamate in three types of Chinese wines and its possible reasons

A total of 75 wines including 30 white wines, 31 red wines, and 14 sparkling wines were obtained from several regions in China (Sinkiang, Tonghua, Huailai, Yantai, Changli, Shanxi, Gansu, and Ningxia). Ethyl carbamate (EC) was detected by gas chromatography mass spectrometry. The EC concentration ranged from less than 1.16 to 38.56 μg/L, and the concentrations in 17 wines exceeded the U.S. limit for table wines (15 μg/L). The concentrations of EC increased in the order of white, red, and sparkling wines with the corresponding mean concentrations of 6.12, 9.22, and 14.03 μg/L. The relationship between EC concentration and wine type suggested that EC concentrations in wines might be affected by vinification patterns, most likely due to the difference between EC precursors in different vinification processes. This work provides a novel clue for EC contamination in different wines.     

新疆焉耆盆地产区4种甜白葡萄酒中挥发性香气成分分析

为了研究焉耆盆地产区甜白葡萄酒的挥发性香气成分,延伸该产区甜白葡萄酒的产业链,该实验采用顶空固相微萃取（HSSPME）结合全二维气相色谱-飞行时间质谱（GC×GC-TOFMS）检测了新疆焉耆盆地产区的4种甜白葡萄酒中的香气成分,并对分析结果进行主成分分析（PCA）。结果表明,新疆焉耆盆地产区4种甜白葡萄酒中共检测出103种香气成分,其种类相似,但含量有差别,酯类和醇类物质总含量在各样品中均占80%以上。通过PCA初步确定了焉耆盆地产区4种甜白葡萄酒中各自的特征香气化合物,且4种甜白葡萄酒挥发性香气成分之间的得分关联度较小,构成了甜白葡萄酒独特的风味特征。     

Distinguishing organic wines from normal wines on the basis of concentrations of phenolic compounds and spectral data

This study was undertaken to determine whether it is possible to distinguish organic wines from normal wines on the basis of chemical composition and spectral data. A total of 58 wine samples including organic and normal red and white wines from France and Germany were analysed. The concentrations of several phenolic compounds, including trans-resveratrol, were determined using reversed-phase high-performance liquid chromatography with both diode array and coulometric electrode array detection. In addition, total phenols, total acids, pH and SO₂ were determined. The ultraviolet-visible (UV-vis) spectra of wines were measured, and the absorbances at four different wavelengths were determined. The data were processed statistically using principal component analysis (PCA). Despite several different combinations of variables, separation of white wines was only achieved according to geographical origin. PCA differentiated well between organic Burgundy red wines and normal Burgundy wines. The separation was based on the concentrations of caffeic and gallic acids, total phenol content and the absorbance ratios A₂₅₄/A₂₇₆, A₂₇₆/A₃₂₀ and A₃₂₀/A₅₂₀. PCA was unable to differentiate organic white wines from corresponding normal wines. The mean trans-resveratrol concentrations of 16 organic red wines and 16 normal red wines were 5.6 mg/l and 2.9 mg/l, respectively. The highest concentration, 14,5 mg/l, was found in an organic red wine from the Loire district in France.     

Volatile profile and differentiation between Albariño wines from different origins

Commercial Albariño wines from different subzones of Certified Brand of Origin ‘Rías Baixas’ (NW Spain) were analysed over four consecutive harvests. The contents of monoterpenes, volatile phenols, alcohols, sulphur compounds, acetates, fatty acids and ethyl esters were determined by gas chromatography (flame ionisation detector and flame photometric detector) and gas chromatography–mass spectrometry. These white wines presented high concentrations of free monoterpenes, volatile phenols, acetates, and ethyl esters, while the levels of sulphur compounds, known as default compounds, were very low. All the wines were elaborated from Albariño grapes, but they were produced by different enological practices, such as skin contact and by carrying out malolactic fermentation or not. No significant differences were found between wines from the same subzone and from different vintages. The results obtained, and confirmed by Principal component and linear discriminant analysis, showed a differentiation among these wines as a function of the Galician subzone of origin.     

Comparative study of phenolic compounds in Cabernet Sauvignon wines made in traditional and Ganimede fermenters

Chemical profiles of anthocyanin and non-anthocyanin phenolics of Cabernet Sauvignon wine made by two different winemaking techniques (traditional vinification and Ganimede method) were determined by high performance liquid chromatography–mass spectrometry (HPLC–MS). Particularly, effect of extraction on and subsequent stability of the phenolic compounds from the end of fermentation to bottling were investigated. The results showed that the total anthocyanin content was higher in the young wines produced in the Ganimede fermenter. The anthocyanin contents in these wines subsequently decreased significantly after two years of ageing. By contrast, the traditional vinification was slightly better than the Ganimede to yield the non-anthocyanin phenolics. This indicates that the Ganimede fermenter might be suitable for the production of brightly coloured red wines for early consumption, which could save time and labour cost for industrial production of highquality wines.     

Changes of Phenolic Acids During Aging of Organic Wines

Grapes (Vitis vinifera L. origin var: Carignane, Cabernet Sauvignon, Merlot, Grenache, Columbard, and Semillon) were cultivated and processed according to accepted organic agriculture and organic wine techniques. Aged wines (5 years) were evaluated for changes of their phenolic acids. The highest reduction of gallic acid concentrations were determined in Cabernet Sauvignon (24.36 mg/L) and Carignane (16.00 mg/L) wines. The quantities of p-hydroxybenzoic acid decreased mostly in Carignane (22.47 mg/L) and Columbard (20.84 mg/L) wines. The decreases of syringic acid were dominant in Cabernet Sauvignon (2.34 mg/L) and Carignane (1.69 mg/L) wines. In the case of ferulic acid, the highest reduction was determined in Cabernet Sauvignon (3.97 mg/L) wines. The decreases of p-coumaric acid contents were the same and mostly in Merlot (1.06 mg/L) and Grenache (1.035 mg/L) wines. The principal component analyses results demonstrated the relations among aged wines produced from different grape varieties and their phenolic acids. The distribution of data was accumulated into two groups. The first group included total phenols, gallic acid, p-hydroxybenzoic acid, syringic acid, p-coumaric acid, and Merlot/Carignane/Grenache wines. The second one included ferulic acid and Cabernet Sauvignon wine.     

Polyphenolic Composition and Antioxidant Activity of Galician Monovarietal Wines from Native and Experimental Non-Native White Grape Varieties

Galicia region (Northwestern Spain) wine production is mainly focused to high-quality white wines. The polyphenolic composition and the antioxidant activity have been determined to characterize the wines obtained from white grape varieties (Albariño, Branco Lexitimo, Caiño blanco, Godello, Loureiro, Torrontes, and Treixadura) grown in Galician protected production areas, and the wines elaborated with non-native varieties experimentally grown in Galicia (Chardonnay, Gewürztraminer, Pinot blanc, Pinot gris, Riesling, and Sauvignon blanc). Since Albariños are the varietal wines with the highest production and commercialization extension, we have studied their polyphenolic composition as a function of the production subarea. Three vintage years (2010, 2011, and 2012) have been considered to account for climate variability. Liquid chromatography with diode array and mass spectrometry detection was used to obtain the phenolic profiles. Wines from native grapes could be fully discriminated by their phenolic composition, with only Albariño being partially confused with other varietal wines. Albariño wines produced in O Rosal and Ribera do Ulla could be clearly differentiated, whereas Condado do Tea and Val do Salnés wines were partially confused. The polyphenolic profile was enough to differentiate the wines obtained from non-native grapes. The total polyphenols content and the antioxidant activity of white wines elaborated in Galicia were comparable, although significant differences were found among varieties.