Detection of ochratoxin A in tropical wine and grape juice from Brazil

BACKGROUND: Ochratoxin A (OTA) is the main mycotoxin found in grapes, wines and grape juices and is considered one of the most harmful contaminants to human health. In this study, samples of tropical wines and grape juices from different grape varieties grown in Brazil were analysed for their OTA content by high‐performance liquid chromatography. RESULTS: The detection and quantification limits for OTA were 0.01 and 0.03 µg L^?1 respectively. OTA was detected in 13 (38.24%) of the samples analysed, with concentrations ranging from < 0.03 to 0.62 µg L^?1. OTA was not detected in any of the grape juice samples. Most of the red wine samples proved to be contaminated with OTA (75%), while only one white wine sample was contaminated. However, the OTA levels detected in all samples were well below the maximum tolerable limit (2 µg L^?1) in wine and grape juice established by the European Community and Brazilian legislature. CONCLUSION: The results of this study indicate a low risk of exposure to OTA by consumption of tropical wines and grape juices from Brazil. © 2012 Society of Chemical Industry     

Ochratoxin A in wines,musts and grape juices from Spain

A survey on the occurrence of ochratoxin A (OTA) in 240 grape‐based beverages was carried out. Red and white wines from four different Spanish Designations of Origin (n = 160), musts (n = 20), grape juices (n = 10), ordinary wines (n = 20), special wines (Malaga, muscatel, sherry, vermouth, etc) (n = 20) and sparkling wines (n = 10) were assayed for OTA content using immunoaffinity column clean‐up and high‐performance liquid chromatography with fluorimetric detection (detection limit 0.05 µg l^?1). Forty‐three (17.9%) of the samples tested contained detectable levels of OTA. The overall mean OTA concentration in red and white wines of Designations of Origin was 0.30 and 0.18 µg l^?1 respectively (ranges 0.05–3.19 and 0.05–1.13 µg l^?1 respectively). The percentage of wine samples with detectable amounts of OTA was higher for red (18.3%) than for white (10%) wines. OTA was also found in two of 10 red ordinary wines (0.68 and 4.24 µg l^?1), whereas none of 10 white ordinary wines contained OTA. The mean OTA amount detected in sparkling wines was 0.44 µg l^?1 (range 0.14–0.71 µg l^?1). Two of 20 must samples contained OTA at low levels (0.08 and 0.18 µg l^?1), while none of 10 grape juice samples contained OTA. Highest amounts of OTA were found in special wines (45%), with a maximum of 15.25 µg l^?1 in a muscatel sample. Copyright © 2004 Society of Chemical Industry     

Survey of sulfites in wine and various Turkish food and food products intended for export, 2007–2010

Surveys were carried out between 2007 and 2010 to determine the total levels of sulfites in 1245 samples of wines, dried apricots, dried vegetables, nuts, juices and purees, frozen foods and cereals containing dried fruit supplied by food inspectors and by food producers for testing or for export certification. Sulfite analysis of wine was carried out using the Ripper method with an LOQ of 5?mg?l^?1 and for dried and other foods the Monier-Williams distillation procedure was employed with an LOQ of 10?mg?kg^?1. In the survey all wines contained measurable sulfites, but with the exception of one sample of white wine they were otherwise below Turkish Food Codex limits of 160?mg?kg^?1 for red wine, 210?mg?kg^?1 to white wine and 235?mg?kg^?1 for sparkling wine. None of the cereal products, frozen foods, juices or purees contained sulfites above 10?mg?kg^?1. However, all dried apricot samples contained significant levels of sulfite with around 40% having levels exceeding the Turkish limit of 2000?mg?kg^?1. Significant levels of sulfite were found in other samples of dried fruit with even a fruit and nut bar containing 1395?mg?kg^?1 of sulfite, suggesting the dried fruit ingredients contained levels above regulatory limits.     

Occurrence and stability of inorganic and organic arsenic species in wines,rice wines and beers from Central European market

We investigated in total 80 wine samples of different types and seven grape juice and 23 beer samples purchased from markets in Central Europe in order to understand the arsenic (As) speciation and help assess the potential As toxicity via intake of alcoholic beverages. Generally, total As concentrations in most samples investigated were below the drinking water limit 10?µg?l^?1 published by the World Health Organization (WHO); ranging from 0.46 to 21.0?µg?l^?1 As in red and white wines and from 0.75 to 13.4?µg?l^?1 As in beers. In addition, concentrations of total As in rice wine and in rice beer were 0.63–6.07 and 3.69–8.23?µg?l^?1 As, respectively. The total As concentrations in ice wine ranged from 7.94 to 18.8?µg?l^?1 As, significantly higher than in white and red wine. Arsenite predominated as the As species in most of the wine samples, whereas arsenate was the dominant species in rice wine, beer and rice beer. Methyl As components were usually minor components in all wine and beer samples. Monomethylarsonic acid, dimethylarsinic acid and two additional unknown As species were frequently found in grape juice, late harvest and ice wine with higher sweetness. After air exposure, arsenite, arsenate, monomethylarsonic acid and dimethylarsinic acid were stable at 4°C for months, probably due to the acidic conditions of wine and beer samples. The presence of sulfite had little influence on As speciation in wine. Despite the predominance of more toxic arsenite and arsenate in wine and beer, the estimated weekly exposure to As (via consumption of beer, wine and rice wine) is low. The As intake per capita is 6.81?µg from beer, <1.93?µg from wine and 0.88?µg from rice wine, estimated using the median of total As concentration multiplied by the average consumption per capita of the corresponding beverage.     

Ochratoxin A in wine and grape juice sold in Canada.

Ochratoxin A (OTA) was determined in 251 samples of wines and grape juice collected over 3 years in Canada. In total, 25/84 samples of red wine, 22/96 samples of white wine, 3/46 red grape juices and 1/25 white grape juices contained OTA levels above the limit of quantitation (LOQ). Canadian wines, when compared with imported products, showed both a lower OTA occurrence, noted as positive (19 versus 48% above the limit of detection (LOD) for wines), and a lower level of OTA contamination (upper bound mean of 17.5 versus 163pg ml(-1) for wines). Wines from the USA contained no quantifiable levels of ochratoxin A. OTA was found in Canadian and US grape juice samples, with 12.9% above the LOD and an upper bound mean of 13.3pg ml(-1). It was extracted from a wine or grape juice sample by passing it through an immunoaffinity column. The sample matrix was washed off the column with water. OTA was eluted from the column with methanol and quantitatively determined by liquid chromatography using a fluorescence detector. The presence of OTA was confirmed by esterification with boron trifluoride-methanol. The LOQ of OTA was estimated as 20 pg ml(-1) in white wine (S/N 10:1) and 40 pg ml(-1) in red wine, white grape juice and red grape juice (S/N 20.1). The LOD was estimated as 4pgml(-1) for white wine and 8pgml(-1) for red wine and white and red grape juices (S/N 3:1).     

Levels of histamine and other biogenic amines in high-quality red wines

Biogenic amines in wine may impair sensory wine quality and cause adverse health effects in susceptible individuals. In this study, histamine and other biogenic amines were determined by HPLC after amine derivatisation to dansyl chloride conjugates in 100 selected high-quality red wines made from seven different cultivars. Amine levels varied considerably between different wines. The most abundant amines were putrescine (median?=?19.4?mg?l^?1, range?=?2.9–122), histamine (7.2?mg?l^?1, 0.5–26.9), and tyramine (3.5?mg?l^?1, 1.1–10.7), whereas lower levels were found for isoamylamine (median?=?0.25?mg?l^?1), phenylethylamine (0.16?mg?l^?1), cadaverine (0.58?mg?l^?1), spermidine (1.8?mg?l^?1) and tryptamine (0.06?mg?l^?1). Positive correlations were observed between isoamylamine and phenylethylamine, and between histamine, putrescine and tyramine levels. Amine concentrations were similar in all wine cultivars except Pinot noir and St. Laurent wines, which showed significantly higher tryptamine and cadaverine levels. The results indicate that levels of histamine and other biogenic amines may vary considerably between red wines independent of grape variety and that high amounts can also be found in high-rated wines. Adopting a legal histamine threshold level of 10?mg?l^?1 in the European Union, as formerly introduced in other countries, would have excluded 34% of the investigated wines from the market.     

Spectral evaluation of total phenolic components in Vitis vinifera: Grapes and wines

Whereas high levels of flavonoid extractives are responsible for the characteristic λ_Dmax at about 280 nm in the ultra-violet spectra of red wines, there is wide variability in the spectral features of natural white wines, in which the phenolic content is often minimal. For the latter, λ_Dmax may occur anywhere within the range 265–285nm, with relatively strong absorbance at 320 nm arising from the presence of hydroxycinnamate esters. Massive fining of many white wines and juices with polyvinylpyrrolidone (10% w/v) has shown residual non-phenolic u.v. absorbing constituents to be sufficiently uniform to enable direct spectral measurement of total hydroxycinnamates in white juices, white wines and rosés as (E₃₂₀-1.4) absorbance units. For white grape juices and white wines, total hydroxycinnamates are directly quantitated as mg litre^?1 ‘caffeic acid equivalents’ and total flavonoid extractives may also be estimated. Total phenolic components of red wines are accounted as (E₂₈₀-4) absorbance units. The spectral interpretations enable comparative estimation of flavonoid extractives in all natural wine types.     

Efficacy of ultraviolet radiation as an alternative technology to inactivate microorganisms in grape juices and wines

Since sulphur dioxide (SO₂) is associated with health risks, the wine industry endeavours to reduce SO₂ levels in wines with new innovative techniques. The aim of this study was, therefore, to investigate the efficacy of ultraviolet radiation (UV)-C (254 nm) as an alternative technology to inactivate microorganisms in grape juices and wines.A pilot-scale UV-C technology (SurePure, South Africa) consisting of an UV-C germicidal lamp (100 W output; 30 W UV-C output) was used to apply UV-C dosages ranging from 0 to 3672 J l⁻¹, at a constant flow rate of 4000 l h⁻¹ (Re > 7500). Yeasts, lactic and acetic acid bacteria were singly and co-inoculated into 20 l batches of Chenin blanc juice, Shiraz juice, Chardonnay wine and Pinotage wine, respectively. A dosage of 3672 J l⁻¹, resulted in an average log₁₀ microbial reduction of 4.97 and 4.89 in Chardonnay and Pinotage, respectively. In Chenin blanc and Shiraz juice, an average log₁₀ reduction of 4.48 and 4.25 was obtained, respectively.UV-C efficacy may be influenced by liquid properties such as colour and turbidity. These results had clearly indicated significant (p < 0.05) germicidal effect against wine-specific microorganisms; hence, UV-C radiation may stabilize grape juice and wine microbiologically in conjunction with reduced SO₂ levels.     

Analysis of Ochratoxin A in Wine by High-Resolution UHPLC-MS

A method for determination of ochratoxin A (OTA) in wines using a new-solid phase extraction clean-up procedure followed with ultra performance liquid chromatography (UHPLC)-Orbitrap MS based on two scan events (full-scan Fourier transform mass spectrometer [FTMS] and higher energy-induced collision dissociation[HCD] data-dependent MS/MS) in positive ionization mode has been developed. The limit of detection (LOD) was estimated at 0.46 μg l^?1 for white wine, 0.53 and 0.54 μg l^?1 for rosé and red wines, respectively. The limit of quantification (LOQ) was estimated at 1.57 μg l^?1 in white wine, 1.77 and 1.81 μg l^?1 in rosé and red wines. Recovery experiments were carried out with spiked samples at three concentration levels (2, 5 and 10 μg l^?1). The OTA recoveries in spiked white wine samples varied from 69.6 % to 99.8 %, while the recoveries for rosé and red wine samples were in the range of 63.0–110.2 % and 63.6–103.2 %, respectively. Finally, based on the results, it is concluded that the combination of C18 cartridge with conventional particle packed columns and UHPLC LTQ-Orbitrap XL is an appropriate procedure for OTA analysis in wines.     

Detection of ochratoxin A and cis- and trans-resveratrol in red wines and their musts from Calabria (Italy)

The natural occurrence of ochratoxin A (OTA) and cis- and trans-resveratrols in red wines has been widely reported. The aim of this work was to evaluate the ochratoxin A (OTA) and both cis- and trans-resveratrol content of red wine (from must to wine) in a pilot-scale vinification process in Calabria (Italy). Eleven samples were collected at different stages of vinification and analysis was carried out by HPLC. Wine from manufacturer 3 contained the highest amount of trans-resveratrol (3.41?mg?l^?1). This wine was characterized by an Aglianico–Magliocco grape variety. Interestingly, data regarding OTA showed that the value of this contaminant was low in all analyzed samples and, in each case, below the legal limit (2.0?mg?l^?1 (ppb)). Overall, the results demonstrated the high quality of wines produced in Calabria.     

Deacidification of red and white wines by a mutant ofSchizosaccharomyces malidevoransunder commercial winemaking conditions

A mutant ofSchizosaccharomyces malidevorans, which utilizes malic acid more rapidly than the wild-type strain and does not utilize significant amounts of glucose or fructose, was used in commercial scale deacidification of grape juice during two vintages in a New Zealand winery. Four trials were conducted in Chardonnay, Semillon and Cabernet Sauvignon juices, 1000–4500 l. In the first two trials, the juices were inoculated with the alcoholic fermentation yeast,Saccharomyces cerevisiae, 33–48 h after the mutant inoculation. In the remaining two trials, the juices were inoculated simultaneously with the mutant andS. cerevisiae. Malic acid, ranging from 3.5–10 g l⁻¹in the juices, was completely degraded within 21–73 h. TheSchiz. malidevoransmutant utilized malic acid at sulfur dioxide levels and temperatures used in red and white winemaking. The wines produced in the trials lacked obvious organoleptic defects. They were blended with untreated wines for acid balance and retailed as varietal wines.     

Degradation of white wine haze proteins by Aspergillopepsin I and II during juice flash pasteurization

Bentonite is commonly used to remove grape proteins responsible for haze formation in white wines. Proteases potentially represent an alternative to bentonite, but so far none has shown satisfactory activity under winemaking conditions. A promising candidate is AGP, a mixture of Aspergillopepsins I and II.; a food grade, well characterized and inexpensive protease, active at wine pH and at high temperatures (60-80°C). AGP was added to two clarified grape juices with and without heat treatments (75°C, 1min) prior to fermentation. AGP showed some activity at fermentation temperatures (≈20% total protein reduction compared to control wine) and excellent activity when combined with juice heating (≈90% total protein reduction). The more heat stable grape proteins, i.e. those not contributing to wine hazing, were not affected by the treatments and therefore accounted for the remaining 10% of protein still in solution after the treatments. The main physicochemical parameters and sensorial characteristics of wines produced with AGP were not different from controls.     

A survey of ochratoxin A occurence in Greek wines

A total of 150 wines, including 123?dry wines (64 red, 49 white and 10 rosé) and 27 dessert wines (14 red and 13 white), were obtained from various viticulture and oenological practices across Greece during the period 1999–2006 and analyzed for ochratoxin a (OTA) using immunoaffinity clean-up and HPLC with fluorescence detection. There was a high frequency of OTA in commercially available wines (69% positive samples). However, the level of contamination was relatively low, with only one sample marginally reaching the EU permitted maximum level (2.0?µg?l^?1). A total of 91% of the samples had OTA concentrations <1.0?µg?l^?1. The higher concentrations were found in wines from the southern regions, especially in dessert-type wines. There were no significant differences based on wine color or production years. Furthermore, there was no difference between conventional or organic cropping systems in terms of OTA presence.     

Liquid chromatographic analysis of maneb and its main degradation product,ethylenethiouera, in fruit juice

Ethylenethiourea (ETU), a possible human carcinogen and an antithyroid compound, is the main degradation product of the fungicide, maneb, which is widely used in agriculture. In this study, a rapid and accurate method for the determination of maneb and ETU in various fruit juices (tomato, grape and apple) was developed requiring minimal clean-up of sample extract, no derivatization prior to injection and no specialized LC detectors. Samples were cleaned up using silica and octadecylsilica (C₁₈) cartridges before injection into liquid chromatography (LC) with diode-array detection (DAD). Recoveries ranged between 90 and 101% with relative standard deviations from 0.7 to 3.8%. The limits of determination of maneb and ETU were 0.1 and 0.01 mg l^?1, respectively. The proposed method was used to monitor the presence of maneb and ETU in commercial samples taken from different markets of Istanbul, Turkey. Maneb was found in one tomato juice sample at a concentration of 0.45 mg l^?1 but ETU was below the LOQ. Two tomato juices had no detectable maneb residue but contained ETU at levels of 0.08 and 0.11 mg l^?1.     

Preliminary studies on acidity-astringency interactions in model solutions and wines

Intensity of astringency in model solutions and wines varying in total acid and total phenols was evaluated by paired comparisons by 10 trained judges. Model solutions consisted of tannic (500, 1000, 2000 mg litre^?1) and tartaric (0, 2, 4, 6 g litre^?1) acids dissolved in aqueous solutions of ethanol (120 ml litre^?1) and sucrose (5 g litre^?1). Wine solutions were prepared by addition of citric acid (0, 0.5, 1, 2, 3 g litre^?1 as tartaric acid) to a high phenol-red wine (2645 mg litre^?1 GAE) and a moderate phenol-white wine (800 mg litre^?1 GAE). At all three levels of tannic acid, astringency of model solutions increased significantly (P<0.001) with tartaric acid concentration. Astringency of white wine also increased significantly (P<0.05) with citric acid concentration. A negative linear relation was found between relative astringency and pH at a given tannin level for both model solutions and white wine. As pH was reduced, more phenolic molecules were in the phenol form thereby increasing the likelihood of hydrogen bonding between hydroxyl groups of wine tannins and ketoimide groups of mouth proteins. The preliminary hypothesis that hydrogen bonding is the main reaction involved in the formation of protein-tannin complexes resulting in the sensation of astringency was reinforced.     

Simple determination of main organic acids in grape juice and wine by using capillary zone electrophoresis with direct UV detection

An accurate, simple and rapid capillary zone electrophoresis (CZE) method with direct UV detection has been set up for the determination of main organic acids in grape juice and wine. The determination of tartaric, malic, and citric acids in grape juices and tartaric, malic, succinic, acetic, lactic and citric acids in wines can be achieved in less than 3 min with only a simple dilution and filtration treatment of the sample. Validation parameters of the method as detection and quantification limits, linearity, precision (intraday and interday analysis) and recovery were also studied in grape juice, white wine, rose wine and red wine, separately. The proposed method decreases the analysis times of the previous reported CZE methods and allows the rapid control of the grape maturity, the winemaking processes and the detection of wine alterations and/or illnesses.     

Colour and label evaluation of commercial pasteurised red juices and related drinks

Despite growing demand by consumers for healthy beverages, artificial colours are still widely used. Levels of anthocyanins and artificial colours were determined by HPLC with UV-Vis detection in red orange juices and other red beverages (nectar, juice-based, health, carbonated and sports drinks). The contribution of pigments to the visible colour of the beverage was calculated. Red orange juice samples contained about 34?mg?l^?1 of anthocyanins, which were responsible for about 92% of the visible colour. Red juice-based drinks, containing from 0% to 30% of red orange, berry, grape or pomegranate juices, had low levels of anthocyanins (about 7?mg?l^?1) and high levels of E129 (about 32?mg?l^?1), which were responsible for about 90.7% of the colour of these beverages. Red health drinks, enriched with vitamins and polyphenols, contained from 3% to 50% of red fruit juices. Also in this case the E129 levels were higher (about 22?mg?l^?1) than anthocyanins (about 9?mg?l^?1), and were responsible for the colour of the beverages (76.1%). High levels of artificial colours were found in red orange carbonated drinks, but in comparable amounts with those found in the other beverage samples, while anthocyanins were only present in trace amounts. Although all of the beverages claimed to contain red fruits on the labels, no correlation was found between the level of anthocyanins and the declared percentage of red fruits. These labels generally conformed with the requirements of the law, but food product labels can often be misleading to consumers about the real characteristics of the product.     

Statistical optimization of ethanol fermentation parameters for processing local grape cultivars to wines

Grape juice from two local grape cultivars viz. Punjab MACS purple and H‐144 was subjected to prefermentation skin treatment. Ethanol fermentation w.r.t agitation rate, temperature, inoculum size, and nutrient supplementation were optimized using Triple M medium following response surface methodology (RSM). RSM results were numerically optimized keeping temperature “in range” for red wine and “low” for white wine which showed agitation rate of 80 ± 1 rpm for 24 hr, diammonium hydrogen orthophosphate supplementation @ 150 mg/100 ml, inoculum size of 6.1 and 6.5% (vol/vol), and fermentation temperature of 24.6 and 21 °C as optimum for ethanol fermentation of red and white wines, respectively. Optimized results were validated on grape juice of Punjab MACS purple and H‐144 cultivars that lead to 12.0 and 11.2 (%vol/vol) ethanol production, respectively. Gas chromatography mass spectrometry analysis revealed the presence of 41 volatile compounds in the form of phenols, alcohols, terpenes, esters, ketones, and amines that add to the nutraceutical and antioxidant value of the wines.

Practical applications

Present study provides the statistical optimized fermentation parameters for red and white wine production separately. Optimization of an efficient processing technology to produce local grape wines will help to reduce the price of wines so that they are available to common masses at affordable costs besides improving the economic status of grape growers in the state and providing valuable information to wine makers to establish winery under North Indian conditions. Use of synthetic grape juice (Triple M media; during off season of grapes) lead to optimization of ethanol fermentation parameters in two separate fashions considering fermentation temperature as key parameter. Gas chromatography mass spectrometry analysis showed the presence of flavonoids, terpenes, and esters increasing nutritive value of product, providing antioxidants to the consuming person.     

Evaluation of the antiradical and reducing properties of selected Greek white wines: correlation with polyphenolic composition

The total hydroxycinnamate (THC) and total flavanol (TF) contents were determined in 26 selected Greek white wines of ‘Appellation of Origin of High Quality’ using well‐established spectrophotometric methodology. Furthermore, two parameters related to the antioxidant capacity of wines, the antiradical activity (A_AR) and the reducing power (P_R), were also determined using the stable DPPH· radical and a ferric reducing power assay respectively. THC content varied from 64.4 to 197.1 mg l^?1 chlorogenic acid equivalents (CGAE), whereas TF content exhibited larger variation, ranging from 3.3 to 205.3 mg l^?1 catechin equivalents (CTE). A_AR and P_R were found to range from 0.47 to 0.60 mM Trolox equivalents (TRE) and from 0.32 to 2.05 mM quercetin equivalents (QE) respectively. Regression analysis at 95% significance level indicated that reducing effects are likely to be exerted by the flavanol fraction (SF < 0.05), as opposed to antiradical efficiency which is primarily affected by the sum of THC and TF contents (SF < 0.05), suggesting that synergistic phenomena account for the overall antioxidant status of white wines. © 2002 Society of Chemical Industry     

Anthocyanin profile in berry skins and fermenting must/wine, as affected by grape ripeness level of Vitis vinifera cv. Shiraz/R99

The release of anthocyanin compounds from berry skins into the wine is affected by several viticulture and oenological practices. Ripeness level seems to play a significant role. The study aimed to evaluate the extraction kinetics of individual anthocyanins from grape skins into wine under controlled conditions of small-scale vinification of Shiraz grapes, harvested at three ripeness levels (23, 25 and 28°Brix). The anthocyanin profile of intact berry skins, fermenting musts/wines and crushed skins during the period between crushing and pressing was analysed by HPLC. Ripeness level greatly impacted on the skin?/?juice ratio (ranging from 169?± 4.5?g?L^?1 at 23°B, 185?±?7.2?g?L^?1 at 25°B and 256?±?10.7?g?L^?1 at 28°B) and on grape anthocyanin concentration (ranging from 0.80?±?0.13?g?kg^?1 at 23°B, 0.78?±?0.07?g?kg^?1 at 25°B and 1.21?±?0.13?g?kg^?1 at 28°B) and profile (%) at harvest. In addition, it affected the rate and amount of individual anthocyanin extraction from skins into wine and their transformation during fermentation, consistent with the relevant chemical group classification (3-glucoside, 3-acetyl-glucoside, 3-p-coumaroyl-glucoside). At pressing, the anthocyanin concentration of the three wines was similar (319?mg?L^?1, on average), but the anthocyanin profile was different, particularly the ratio of 3-glucoside?/?3-p-coumaroyl-glucoside derivatives, which was on average 3.55 at 23°B and 25°B and 2.6 at 28°B. Viticultural choices, such as to harvest earlier or later, influencing the chemical and physical composition of the berries, may influence the extraction kinetics of individual anthocyanins and their destiny in the fermenting must, offering to winemakers different basic wines suitable for the production of different wine products. These findings are valuable to improve viticultural and oenological practices for Shiraz wine production, allowing the improvement of wine quality and the purposeful creation of different styles of wine.