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.     

Natural co-occurrence of ochratoxin A,ochratoxin B and aflatoxins in Sicilian red wines

The natural occurrence of ochratoxin A, ochratoxin B, aflatoxin B₁, aflatoxin B₂, aflatoxin G₁ and aflatoxin G₂ (OTA, OTB, AFB₁, AFB₂, AFG₁, AFG₂) in red wines was investigated by HPLC/FLD after immunoaffinity column clean-up in 57 market samples produced in Sicily (Italy). The results showed a very low incidence of these mycotoxins in analysed samples, confirming the high degree of quality and safety of Sicilian red wines. The results indicated 71.9% and 64.9% positive samples for OTA and OTB respectively, with an average level of 0.13 μg l^–1, well below the European maximum permitted levels (MLs). The aflatoxin most frequently detected in the samples was AFG₁, present in 57.9% of samples, while the other aflatoxins were rarely present. Recovery experiments were carried out on eight mycotoxin-free red wines spiked with OTA, OTB, AFB₁, AFB₂, AFG₁ and AFG₂ at two different levels. The limits of detection (LODs) in wines were 0.02 µg l^–1 for OTA, 0.04 µg l^–1 for OTB, 0.03 µg l^–1 for AFG₁, AFG₂ and AFB₂, and 0.05 µg l^–1 for AFB₁. A good correlation was found, with good performances in term of precision for the method.     

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     

Occurrence of ochratoxin A (OTA) in wines: influence of the type of wine and its geographical origin.

The results of more than 450 samples taken from the literature and 400 samples tested by our own investigations have been taken into account to quite extensively describe the situation of OTA contamination of wine. According to these data OTA is much more commonly detected in red wines than in rosé and white wines and the OTA concentration is remarkably higher than in the last two. Thus OTA could be detected in 25% of white wine samples whereas it was detected in 40% of rosé and 54% of red wine samples. The same result was found when comparing the wines from southern and northern regions. Here the red wine samples from the northern cultivating area showed a contamination of 12% in contrast to those from the southern area which showed a contamination of about 95%.     

Occurrence of Ochratoxin A in Chinese wines: influence of local meteorological parameters

A total of 119 wines, including 115 dry wines (77 red, 34 white, 4 rosé) and 4 ice wines, were obtained from four major regions around China (east, northeast, west, and northwest) from 1999 to 2009. Ochratoxin A (OTA) was detected by solid-phase extraction and high-performance liquid chromatography with fluorescence detection. The OTA levels ranged from <0.1 to 5.65 μg/L. The wines from the northeast area had the highest OTA level, with an average concentration of 2.18 μg/L. Analysis of the correlation between meteorological parameters and the OTA concentration in red wine revealed that the OTA concentration was positively correlated with rainfall in August, but negatively correlated with the average temperature. No significant correlation was found with the daily range of temperature and relative humidity. The results show that climate factors affect the OTA distribution in wines from the four major regions of China.     

Free radical scavenging capacity of selected red,rosé and white wines

The free radical scavenging capacity of selected red, rosé and white Spanish wines from different grape varieties was determined by the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH^·) method using a new methodology developed at our laboratory. The amount of sample necessary to decrease by 50% the initial DPPH^· concentration (EC₅₀), the time needed to reach the steady state at EC₅₀ concentration (T_EC50) and the antiradical efficiency (AE = 1/EC₅₀T_EC50) were determined in the wine samples. Some differences between rosé wines made with Garnacha and Tempranillo grape varieties were observed in the UV‐vis spectra and in the free radical scavenging parameters, those from Garnacha variety having the highest antioxidant activity. The antiradical efficiency of the wines followed a decreasing order: red wines (22.44 × 10⁻⁶) > rosé wines (4.90 × 10⁻⁶) > white wines (1.88 × 10⁻⁶). There was a correlation between antiradical efficiency and total polyphenol (TP): AE = −3.33135 + 0.0180535TP; the correlation coefficient was r = 0.951454. © 1999 Society of Chemical Industry     

Ochratoxin A in red pepper flakes commercialised in Turkey

The aim of this study was to investigate the presence of Ochratoxin A (OTA) in red pepper flakes commercialised in Turkey. A total of 75 samples were collected from different supermarkets and traditional bazaars in Istanbul during 2012–2013. OTA analysis was performed by high-performance liquid chromatography equipped with fluorescence detection after immunoaffinity column clean-up. The method was linear in the range 0.05–40 μg kg^?1 (r² = 0.9997). Twenty-seven out of 31 (87.1%) packed red pepper flake samples contained OTA at concentrations ranging from 0.6 to 1.0 μg kg^?1, whereas 100% of the unpacked red pepper flake samples contained OTA, in the range 1.1–31.7 μg kg^?1. Overall, only 4 unpacked samples (5.3%) contained OTA, with a mean value of 23.4 μg kg^?1, which is higher than the European Union limit. It is suggested that OTA content should be carefully considered in red pepper flake samples especially marketed in unpacked form.     

Molecularly Imprinted Nanosilica Solid-Phase Extraction for Bisphenol A in Fish Samples

This paper reports a method using molecularly imprinted nanosilica as the solid-phase extraction sorbent to extract bisphenol A (BPA) from fish samples. The selective extraction efficiency of BPA from its structure-analogous by molecularly imprinted solid-phase extraction (MISPE) was compared with commercial C18 SPE column. The reproducibility of MISPE and optimal flow rate of sample were studied. There was a linear correlation in the concentration range of 0.7–114.1 μg l^?1 of BPA (r?=?0.997). The limit of detection (LOD) based on three times ratio of signal to noise was 0.11 μg l^?1. Under optimal conditions, the proposed method was applied to the analysis of BPA in fish samples. The amount of BPA in bream was 4.00 μg kg^?1, and both concentrations of BPA in crucian and weever were below the LOD. The recoveries of BPA standard solution spiked with fish samples were in the range of 92.56–102.3 % with the relative standard deviation less than 10 %.     

Impact of wine manufacturing practice on the occurrence of fining agents with allergenic potential

Proteinogenic wine fining agents are hidden allergens and could present a risk for consumers with allergies. Therefore, the European Parliament adopted Directive 2003/89/EC amending Directive 2000/13/EC to declare ingredients, contaminations and processing aids that are known to trigger allergic reactions. The Amendment Regulation (EU) 1266/2010 excluded the labelling of wines which are processed with hen’s egg and products thereof until 30 June 2012 to get more scientific findings. After 1 July 2012 wine fining agents have to be declared if above 0.25 mg l^–1 (Regulation (EU) 579/2012 in conjunction with article 120 g of Regulation (EU) 1234/2007). The Organisation International de la Vigne et du Vin (OIV) advises this limit of detection (LOD) for potential allergenic residues of proteins. Wine fining agents are processing aids and according to the wine producer’s knowledge will be removed after coagulation by filtration or other production steps. Due to lack of scientific data, residues of fining agents in the final product could not be excluded. In this risk assessment, highly sensitive ELISA methods for ovalbumin of known origin for wine have been developed. The objective was to investigate the presence of allergen residues in wine after certain technological treatments were applied to remove the wine fining agents. For all developed ELISA methods the LODs are in the low µg l^–1 range between 5 and 10 µg l^–1 fining agent, whereas the LOQ varies between 5 and 80 µg l^–1 fining agent. The results of the investigation of well-known wines and fining agents demonstrate that white wines fined with white or ovalbumin from hen’s egg could retain allergens. The use of certain technological procedures during wine processing leads to different results. In white wine, bentonite or sheet filtration followed by sterile filtration lead to wines containing no detectable amounts of ovalbumin. In red wine, especially the final sterile filtration removes the fining agents.     

Improved sensitivity of ochratoxin A analysis in coffee using high-performance liquid chromatography with hybrid triple quadrupole-linear ion trap mass spectrometry (LC-QqQLIT-MS/MS)

A novel and sensitive method utilising high-performance liquid chromatography coupled to triple quadrupole-linear ion trap mass spectrometry (LC-QqQLIT-MS/MS) was developed in order to analyse the content of ochratoxin A (OTA) in coffee samples. The introduction of the triple-stage MS scanning mode (MS³) has been shown to increase greatly sensitivity and selectivity by eliminating the high chromatographic baseline caused by interference of complex coffee matrices. The analysis included the sample preparation procedure involving extraction of OTA using a methanol–water mixture and clean-up by immunoaffinity columns and detection using the MS³ scanning mode of LC-QqQLIT-MS/MS. The proposed method offered a good linear correlation (r² > 0.998), excellent precision (RSD < 2.9%) and recovery (94%). The limit of quantification (LOQ) for coffee beans and espresso beverages was 0.010 and 0.003 µg kg^?1, respectively. The developed procedure was compared with traditional methods employing liquid chromatography coupled to fluorescent and tandem quadrupole detectors in conjunction with QuEChERS and solid-phase extraction. The proposed method was successfully applied to the determination of OTA in 15 samples of coffee beans and in 15 samples of espresso coffee beverages obtained from the Latvian market. OTA was found in 10 samples of coffee beans and in two samples of espresso in the ranges of 0.018–1.80 µg kg^?1 and 0.020–0.440 µg l^?1, respectively. No samples exceeded the maximum permitted level of OTA in the European Union (5.0 µg kg^?1).     

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.     

A Sensitive Method for the Rapid Determination of Underivatized Ethyl Carbamate in Fortified Wine by Liquid Chromatography-Electrospray Tandem Mass Spectrometry

This work presents the optimization of a miniaturized liquid-liquid extraction (mLLE) followed by reversed-phase liquid chromatography-electrospray tandem mass spectrometry (RP-HPLC-MS/MS) for the determination of ethyl carbamate (EC) in fortified wine, without using derivatizing agents. The mLLE was optimized by an experimental design. Thus, 15 mL of wine and 8 mL of ethyl acetate were used for the extraction. After concentration, each extract was injected into the HPLC-MS/MS equipment and the characteristic secondary ion transition of EC (m/z = 90.10 → 62.05) was used for the quantification purposes. The proposed method presented a good linearity (R ² = 0.9999) and a high sensitivity with low limits of detection (LOD) and quantification (LOQ), 0.17 and 0.52 μg L^?1, respectively. The precision (repeatability and reproducibility) never exceeded 8% of variation, and the recoveries varied between 93 and 114%. The applicability of the method was checked through the analysis of 24 fortified wines, with EC values ranging between 23 ± 1 and 194 ± 5 μg L^?1. All chromatograms revealed good peak resolutions. This new method is efficient for the simple, fast, and reliable determination of EC in fortified wines, providing great sensitivity without using derivatizing agents or large volumes of organic solvents.     

Physicochemical and sensory characteristics of red wines from the rediscovered autochthonous Tintilia grapevine grown in the Molise region (Italy)

Tintilia is an autochthonous grapevine of the Italian Molise region which risked to disappear. However, recently, the production of Tintilia red wines is resuming and in the year 2011 the protected designation origin ‘Tintilia del Molise’ was officially registered. In this work, an analytical characterization of representative red wines from Tintilia grape is reported. A total of 36 different physicochemical variables were determined and discussed, considering those with an estimated coefficient of variation <25 % as more characterizing. These were found to be (mean): density (0.9949); dry extract (34.4 g L^?1) and ashes (3.8 g L^?1); ethyl alcohol (14.2 mL 100 mL^?1), glycerol (9.2 g L^?1) and total higher alcohols (1.7 g L^?1); pH (3.65); the titratable (5.9 g L^?1), fixed (5.4 g L^?1), and salified (2.5 g L^?1) acidity; buffering capacity (52.6 mM/L/pH); total phenols (2,341 mg mL^?1); total flavonols (223 mg mL^?1) and epicathechin (75.0 mg mL^?1); %Red (49.1 %) and %Yellow (43.6 %). Sensory analysis was also performed by professional wine tasters. Finally, the Tintilia results were compared with those of Montepulciano wines. Findings of this analytical study describe the Tintilia red wine as a full-bodied wine; alcoholic; with feeble, but stable acidic profile; rich of phenols, especially flavonols; and finally, with a color balanced between red and yellow pigments.     

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     

Simultaneous Determination of Multiclass Pesticides and Antibiotics in Honey Samples Based on Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

A simple, fast, and efficient method was developed for simultaneous determination of 79 pesticides and 13 antibiotics compounds of different chemical classes of pesticides and antibiotics in honey samples by ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). The sample preparation procedure includes homogenization with McIlvaine buffer 0.1 mol L^?1 (pH 4), followed by extraction with acetonitrile and cleanup with florisil, using dispersive solid phase extraction (d-SPE). The proposed method was validated with good results, such as linearity (r ²?>?0.9901), normality, and independence of the evaluated data, as well as recoveries between 70 and 120 % with relative standard deviation (RSD) <20 % for most of the compounds spiked from 0.1 to 200 μg kg^?1. The experimental method limits of detection and quantification were from 0.03 to 1.51 μg kg^?1 and from 0.1 to 5 μg kg^?1, respectively, for the pesticides. For the antibiotics, the decision limits (0.1 to 2 μg g^?1) and the detection capacity (0.12 to 2.81 μg g^?1) were below the maximum residue limits (MRLs) established for honey by the Brazilian and European legislation. The method was successfully applied to real samples from different botanical and geographic origins. From them, 44 % presented residues from 0.12 to 10 μg kg^?1 of one or more analytes. The proposed method combines the advantages of a quick sample preparation step with the selectivity and sensitivity of the UHPLC-MS/MS and proved to be suitable for routine analyses.     

Tocopherols and Tocotrienols: an Adapted Methodology by UHPLC/MS Without Sample Pretreatment Steps

Ultra high-performance liquid chromatography-electrospray ionization-mass spectrometry (UHPLC/ESI/MS) methodology was adapted for identification and quantification of tocopherols and tocotrienols in vegetable oils with no derivatization or sample preparation steps. The UHPLC analysis was performed using a C18 column and mobile phase composed of methanol: water: ammonium hydroxide (99:1:0.1 v/v/v) and isopropanol. A single mass spectrometer with electrospray on negative mode was used as a detector for tocopherols and tocotrienols. The samples were diluted in isopropanol. The limit of quantification for tocopherols was 0.006 μg mL^?1, and the linear range was 0.006 to 0.01 μg mL^?1; for tocotrienols, the limit of quantification was 0.002 μg mL^?1, and the linear range of analysis was 0.002 to 0.003 μg mL^?1. The correlation coefficients were higher than 0.99, indicating that the method has suitable linearity. The methodology has proven to be precise, reproducible, and robust for the parameters studied.     

Co-occurrence of aflatoxins,ochratoxin A and citrinin in “egusi” melon (Colocynthis citrullus L.) seeds consumed in Ireland and the United Kingdom

The natural co-occurrence of aflatoxins (AFs), ochratoxin A (OTA) and citrinin (CIT) in melon seed samples obtained from retailers and households in Ireland and the United Kingdom (UK) was evaluated. AFs and OTA were determined by HPLC with fluorescence detection while CIT was analysed by HPLC-MS/MS. AFB₁ was detected in all (100%) samples (mean = 9.7 μg kg^?1; range = 0.2–66.5 μg kg^?1). Mean total AFs was 12.0 μg kg^?1 (range = 0.3–82 μg kg^?1). Commercially retailed samples showed a significantly higher AFB₁ contamination (p < 0.05) than the household samples. OTA occurred in 3 (13.6%) samples, while 4 (18.2%) were contaminated with CIT at very low levels. In this study, 68% of the melon seed samples were contaminated above the 2 μg kg^?1 EU limit for AFB₁ in oilseeds. These results highlight the need for the development of strategies to reduce AF contamination in “egusi” for human consumption.     

Development of a Competitive Indirect Enzyme-Linked Immunosorbent Assay for Screening Phenylethanolamine A Residues in Pork Samples

Phenylethanolamine A (PEA), a new alternative β-agonist, has been illegally used in farming to promote the muscle growth in food-producing animals. In this study, a sensitive and convenient competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed for determination of PEA residues in pork samples. The produced antibody was highly specific to PEA and exhibited a negligible cross-reactivity toward some other β-agonists. The developed technique was characterized by the limit of detection below 0.08 μg kg^?1 and the IC₅₀ value of 0.93 pmol mL^?1 (0.32 ng mL^?1). Validation of the technique was done using artificially spiked and naturally contaminated pork samples. The recoveries ranged from 79.6 to 112.6 % for the samples spiked at levels of 0.1–5 μg kg^?1 with the variation coefficients below 15 %. The analysis of naturally contaminated samples showed that the obtained data corresponded with the data obtained by the LC-MS/MS. The developed ciELISA was shown to be a feasible highly sensitive and specific screening tool for PEA residue analysis.     

Determination of Pesticide Residues in Golden Berry (<Emphasis Type="Italic">Physalis peruviana</Emphasis> L<Emphasis Type="Italic">.</Emphasis>) by Modified QuEChERS Method and Ultra-High Performance Liquid Chromatography-Tandem Quadrupole Mass Spectrometry

A fast and effective multiresidue method for the determination of 42 pesticides in golden berry was developed and validated. A modified QuEChERS method was established for sample preparation followed by ultra-high performance liquid chromatographic-tandem mass spectrometry (UHPLC-MS/MS) determination with electrospray ionization in a triple quadrupole system. Validation results were satisfactory, since the method presented recoveries between 70 and 114 % with relative standard deviations (RSD) <20 % for blank samples spiked from 5 to 25 μg kg^?1. The method limit of detection and limit of quantification were 1.5 and 5 μg kg^?1 _, respectively. Matrix effect ranged from ?32 to 218 % and was compensated using matrix-matched calibration. Method linearity was established from 2.5 to 100 μg kg^?1 with r ² ≥ 0.99. The proposed method combines the advantages of a simple and fast sample preparation step by a modified QuEChERS method with the high selectivity and sensitivity of the UHPLC-MS/MS system using selected reaction monitoring. The method was successfully applied to commercial samples, proving to be an efficient alternative for routine analysis. From the 16 analyzed samples, 13 presented residues of one or more pesticides (carbendazim, chlorpyrifos ethyl, dimethoate, propamocarb, and tebuconazole) in the concentration range of 2.0 to 55.6 μg kg^?1.     

Determination of Eugenol in Aquatic Products by Dispersive Solid-Phase Extraction and Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry

A novel procedure, dispersive solid-phase extraction coupled with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), was developed for the determination of eugenol in aquatic products (shrimp, crab, and carp). Aquatic products were extracted with acetonitrile and primarily purified by dispersive solid-phase extraction with graphitized carbon black as absorbent. The pretreated acetonitrile extract was detected by UHPLC-MS/MS. UHPLC was carried out on Dikma Endeavorsil C₁₈ (30 mm × 2.1 mm, 1.8 μm) column eluted by methanol and water (80:20 v/v) at a rate of 0.30 mL min^?1. Tandem mass spectrometry was performed by electrospray ionization in negative ion mode to identify and quantify eugenol during multiple reaction monitoring. Under optimized analytical conditions, the matrix-matched spiked calibration sample demonstrates good linearity between 5.0 and 500.0 μg kg^?1 with a linear regression coefficient of 0.9996. The average recovery of eugenol from aquatic products is 95.3–103.4% at spiked levels between 5 and 50 μg kg^?1 with a relative standard deviation (n = 6) less than 5.4%. The limits of detection and quantification for eugenol were calculated to be 1.47 and 4.91 μg kg^?1, respectively. In comparison with those reported, the proposed method has advantages in low detection limit, high recovery, and short analysis time, meeting the requirements for the determination of trace eugenol residue in aquatic products.