Ochratoxin A survey in Portuguese wine by LC-FD with direct injection

Wine and grape juices were identified as one of the most important sources of ochratoxin A (OTA), a mycotoxin with diverse toxic effects that naturally appears in food and foodstuffs all over the world.The aim of this study was to assess the OTA levels in Portuguese wines through the application of a simple and accurate method based on liquid chromatography (LC) with direct injection, followed by fluorescence detection (FD).Randomly selected wine samples were used to evaluate the performance of direct injection as efficient, fast, inexpensive and safe sample preparation method. The proposed method was successfully validated. The limit of quantification (LOQ) was 1.0 μg/L and OTA recoveries from wine samples, spiked at the three fortification levels, were higher than 85.4%, with RSDs lower than 9.6% for both red and white wines. The presence of OTA was confirmed by methyl ester derivatization followed by LC analysis.Data on OTA levels were obtained for 60 Portuguese red and white wine samples. OTA was found in 12 samples, nine (26%) red wine samples and three (12%) white wine samples. Only one red wine sample and one white wine sample presented a contamination level above the LOQ, with 1.23 and 2.4 μg/L, respectively. It should be pointed out that this white wine sample exceeded the EC maximum permitted level of 2.0 μg/L. The safe dose established as 120 ng/kg body weight/week was not exceeded by the weekly intake estimated for the samples contaminated above the LOQ.     

Potential of supramolecular solvents for the extraction of contaminants in liquid foods

Amphiphile-based supramolecular solvents (ASSs), which are water immiscible liquids consisting of supramolecular aggregates in the nano- and micro-scale regimes dispersed in a continuous phase, were assessed for the extraction of trace contaminants in liquid foods. The ASS selected was made up of reversed micelles of decanoic dispersed in tetrahydrofuran (THF)-water and the contaminants used as a model were bisphenol A (BPA), ochratoxin A (OTA) and benzo(a)pyrene (BaPy). The influence of matrix components on the extractant solvent production, extraction recoveries and actual concentration factors was investigated by using commercial foods such as wine and wine-based products, beer, soft drinks and tea and coffee brews, and/or aqueous synthetic solutions containing specific food matrix components. The method involved the addition of decanoic acid (80mg) and THF (0.8-1.7mL) to the food sample (15mL), stirring of the mixture for 5min, centrifugation for 10min and analysis of 10-20microL of the extract by liquid chromatography coupled to fluorimetry for OTA and BaPy or to mass spectrometry for BPA. No clean-up of the crude extracts was required for any of the samples analysed. The quantification limits for the contaminants (14-31ngL(-1), 0.37-0.39ngL(-1) and 562-602ngL(-1) for OTA, BaPy and BPA, respectively) were far below their respective European legislative threshold limits. Recoveries for food samples were in the ranges 79-93%, 90-96% and 78-82% for OTA, BaPy and BPA, respectively, with relative standard deviations ranging from 1 to 7%, and actual concentrations factors between 65 and 141. The methods developed were applied to the determination of the target compounds in a variety of commercial foods. OTA was found in vinegar, must and beer samples, the concentrations ranging from 92 to 177ngL(-1), BaPy was quantified in samples of tea and coffee at concentrations between 1.5 and 16.6ngL(-1) whereas BPA was detected in two canned soft drinks and quantified in one of them (tea beverage) at a level of 2.3microgL(-1).     

Determination of ochratoxin A in wine by means of immunoaffinity column clean-up and high-performance liquid chromatography

A new and accurate method to quantify ochratoxin A (OA) in table wine has been developed. The method uses commercial immunoaffinity columns for clean-up and high-performance liquid chromatography (HPLC) with fluorescence detection for quantification of the toxin. Wine was diluted with a solution containing 1% polyethylene glycol (PEG 8000) and 5% sodium hydrogencarbonate, filtered and applied to an OchraTest immunoaffinity column. The column was washed with a solution containing sodium chloride (2.5%) and sodium hydrogencarbonate (0.5%) followed by water. OA was eluted with methanol and quantified by reversed-phase HPLC with fluorometric detection (excitation wavelength 333 nm, emission wavelength 460 nm) using acetonitrile-water-acetic acid (99:99:2) as mobile phase. Average recoveries of OA from white, rosé and red wine samples spiked at levels from 0.04 to 10 ng/ml ranged from 88% to 103%, with relative standard deviations (RSDs) between 0.2 and 9.7%. Detection limit was 0.01 ng/ml based on a signal-to-noise ratio of 3:1. The method was applied successfully to 56 samples of red (38), rosé (8), white (9) and dessert (1) wine. The levels of OA ranged from <0.01 to 7.6 ng/ml with red wines more contaminated than rosé and white wines. A good correlation (r=0.987) was found by comparative analysis of 20 naturally contaminated samples using this method and the method of Zimmerli and Dick with better recoveries of OA and better performances for the new method. Several advantages of this method with respect to the actually available methods have been pointed out, with particular reference to red wine which appears to be the most difficult to analyze.     

Quantification of polyphenols with potential antioxidant properties in wines using reverse phase HPLC

A RP-HPLC method with photodiode array detection (DAD) was developed to separate, identify and quantify simultaneously the most representative phenolic compounds present in Madeira and Canary Islands wines. The optimized chromatographic method was carefully validated in terms of linearity, precision, accuracy and sensitivity. A high repeatability and a good stability of phenolics retention times (< 3%) were obtained, as well as relative peak area. Also high recoveries were achieved, over 80.3%. Polyphenols calibration curves showed a good linearity (r(2) >0.994) within test ranges. Detection limits ranged between 0.03 and 11.5 microg/mL for the different polyphenols. A good repeatability was obtained, with intra-day variations less than 7.9%. The described method was successfully applied to quantify several polyphenols in 26 samples of different kinds of wine (red, rosé and white wines) from Madeira and Canary Islands. Gallic acid was by far the most predominant acid. It represents more than 65% of all phenolics, followed by p-coumaric and caffeic acids. The major flavonoid found in Madeira wines was trans-resveratrol. In some wines, (-)-epicatechin was also found in highest amount. Canary wines were shown to be rich in gallic, caffeic and p-coumaric acids and quercetin.     

Coacervative extraction of phthalates from water and their determination by high performance liquid chromatography

A simple, rapid and low cost method for determination of phthalic acid esters (PAEs) including Dimethyl phthalate (DMP), Diethyl phthalate (DEP), Di-n-butyl phthalate (DBP) and Butylbenzyl phthalate (BBP) in water samples was investigated. The method is based on the extraction of PAEs with coacervate made up of decanoic acid reverse micelles and the subsequent determination by HPLC-UV. Effect of parameters such as concentration of tetrahydrofuran (THF) (2?C40% v/v) and decanoic acid (20?C400 mg in 40 ml total volume), ionic strength (0.0?C0.1 M NaCl), pH (1?C4) and stirring time (2?C60 min) on recoveries (Rs) and enrichment factors (EFs) were investigated and optimized. The optimum condition for extraction was the stirring of 36 ml of water sample with 4 ml of THF containing 100 mg of decanoic acid for 10 min and its centrifugation (10 min, 3500 rpm). Recoveries and enrichment factors of PAEs mainly depended on the amount of decanoic acid and THF making up the coacervate and were not affected by ionic strength of the sample solution (up to 0.1 M of NaCl), pH (1?C4), and stirring time (2?C60 min). Recoveries, enrichment factors, LODs and relative standard deviations (RSD%) for PAEs were between 87?C94%, 187?C202, 0.22?C0.30 ??g l^?1 and 2?C5%, respectively. This method was applied to determine PAEs in tap water, river water, and sea water samples. No PAEs were found in tap water. The amount of DMP and DEP in the Babolrood River was 0.87 and 0.67 ??g l^?1, while in the Caspian Sea was 0.49 and 0.52 ??g l^?1, respectively.     

Solid-phase extraction of ochratoxin A from wine based on a binding hexapeptide prepared by combinatorial synthesis

In this paper we describe the preparation of a hexapeptide library by combinatorial synthesis and the identification of a peptide with sequence Ser-Asn-Leu-His-Pro-Lys, which showed good affinity (K(eq)=3.4 x 10(4) M(-1)) towards the mycotoxin ochratoxin A (OTA). An immunoaffinity-like stationary phase supporting such a hexapeptide was used to develop a solid-phase extraction method for the quantification of OTA in wine samples at concentration levels down to 0.10 microg l(-1). Several different wine samples fortified with OTA at 2 and 4 microg l(-1) levels showed recovery of 94.7% and 98.4% at 2.0 and 4.0 microg l(-1), respectively, without any effect on the extraction efficiency of the matrix. The efficacy of this approach was successfully tested by comparison with an immunoaffinity extraction performed on commercial cartridges.     

Determination of volatile phenols in red wines by dispersive liquid-liquid microextraction and gas chromatography-mass spectrometry detection

A new method was developed for analysing 4-ethylguaiacol and 4-ethylphenol in the aroma of red wines using dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-mass spectrometry detection (GC-MS). Parameters such as extraction solvent, sample volume and disperser solvent were studied and optimised to obtain the best extraction results with the minimum interference from other substances, thus giving clean chromatograms. The response linearity was studied in the usual concentration ranges of analytes in wines (50-1500 microg/L). Repeatability and reproducibility of this method were lower than 5% for both volatile phenols. Limits of detection and limits of quantification were also determined, and the values found were 28 and 95 microg/L for 4-ethylguaiacol and 44 and 147 microg/L for 4-ethylphenol, respectively. This new method has been used for the determination of the volatile phenols concentration in different samples of Tannat wine affected by Brettanomyces contamination.     

Occurrence of ochratoxin A in Turkish wines

A total of 95 wine samples including 34 white, 10 rosé and 51 red wines originating from four different Turkish areas were analysed for ochratoxin A (OTA). An analytical method based on immunoaffinity column (IAC) for clean-up and high performance liquid chromatography with fluorescence detection (HPLC-FD) was used to determine OTA in wines. The limit of detection (LOD) was estimated as 0.006 ng ml^− 1 for white wine and 0.010 ng ml^− 1 for rosé and red wines. The limit of quantification (LOQ) was estimated as 0.020 ng ml^− 1 in white wine and 0.030 ng ml^− 1 in rosé and red wines. Recovery experiments were carried out with spiked samples in the range 0.1–1 ng ml^− 1 of OTA. The average OTA recoveries from spiked white wine samples varied from 79.43% to 85.07%; while the mean recoveries for rosé and red wine samples were in the range of 77.48–83.96% and 76.61–83.55%, respectively. OTA was detected in 82 (86%) wine samples at levels of < 0.006–0.815 ng ml^− 1, which were below the maximum allowable limit established by the European Community. The mean OTA concentration in red wines was slightly higher than in white and rosé wines. Furthermore, our data indicate that the geographic region of origin has strong influence on OTA level for white, rosé and red wines: wines originating from Thrace (n = 44, mean = 0.158 ng ml^− 1) and Aegean (n = 28, mean = 0.060 ng ml^− 1) regions of Turkey were more contaminated with OTA compared with wines originating from central (n = 15, mean = 0.027 ng ml⁻¹) and east Anatolia (n = 8, mean = 0.027 ng ml^− 1) areas. This study showed that the occurrence of OTA in Turkish wines is high, but at levels that probably leads to a non-significant human exposure to OTA by consumption of wines.     

Determination of priority carcinogenic polycyclic aromatic hydrocarbons in wastewater and surface water by coacervative extraction and liquid chromatography-fluorimetry

The US Environmental Protection Agency (EPA) and the European Union (EU) have set restrictive limits for priority carcinogenic polycyclic aromatic hydrocarbons (CPAHs) in surface waters (EPA 3.8 ng L(-1) and EU 2-100 ng L(-1)) in order to protect aquatic life and human health. Currently, methods meeting these sensitivity criteria are not suitable for routine analysis of CPAHs. Here, we present a simple, rapid and low-cost method for the routine monitorization of these pollutants in aquatic environments based on their extraction with coacervates of decanoic acid reverse micelles in the nano- and microscale, and determination by liquid chromatography-fluorimetry (LC-FL). The method involves the stirring of filtered aqueous samples (36 mL) with 4 mL of THF containing 70 mg of decanoic acid for 5 min, its centrifugation for 10 min and the analysis of 20 microL of the resulting coacervate containing the CPAHs by LC/FL. The method is robust, the extractions being independent on salt concentration (up to 1 M), temperature (up to 60 degrees C) and pH (below 4). Besides, the coacervate prevents the CPAHs from adsorption onto the surface of containers during sample storage. No clean-up steps are necessary and the method is matrix-independent. The quantification and detection limits of the method ranged between 0.4 and 3.5 ng L(-1) and 0.1 and 1 ng L(-1), respectively, for the seven priority CPAHs. The method has been successfully applied to the determination of these pollutants in raw and treated sewage from three mechanical-biological treatment plants, two rivers and a reservoir with frequent motorized recreational craft activities, all of them located in the South of Spain. Recoveries for spiked samples in the range 2-30 ng L(-1) were between 88 and 95% with relative standard deviations from 1 to 7%. CPAHs were present in wastewater influents at concentrations in the range 3.9-37 ng L(-1), while the treatment at the WWTPs studied reduced their concentration in their respective effluents in a percentage near 100%. Three CPAHs were present at quantifiable levels in Guadajoz river (1.8-6.6 ng L(-1)) and six in La Bre?a reservoir (1.39-4.8 ng L(-1)).     

Exposure assessment to ochratoxin A from the consumption of Italian and Hungarian wines

A total of 267 wine samples including 19 dessert, 186 red, 11 rosé and 51 white produced mostly in the years 1997–2002 in Italian and Hungarian regions were analyzed for ochratoxin A (OTA) using inmunoaffinity column (IAC) clean-up and HPLC with fluorimetric detection. None of Hungarian wine samples were contaminated with this mycotoxin. For Italian red wines, 84% of the samples were positive for OTA ranged from 0.01 to 4.00 ng/mL. Furthermore, OTA was detected in 63% of dessert, in 56% of rosé and in 19% of white wine samples ranged from 0.01 to 1.64, from 0.01 to 1.04 and from 0.01 to 0.21 ng/mL, respectively. A study of OTA daily exposure assessment in Italian wines was also carried out outlining a quite low contribution to the overall daily intake.     

Decanoic acid reverse micelle-based coacervates for the microextraction of bisphenol A from canned vegetables and fruits

Decanoic acid reverse micelle-based coacervates were proposed for the extraction of bisphenol A (BPA) from canned vegetables and fruits prior to its determination by liquid chromatography and fluorescence detection at lambda(exc) = 276 nm and lambda(em) = 306 nm. The procedure involved the extraction of minute quantities (300-700 mg) of homogenized food sample with an aqueous solution containing 10% of THF and 0.5% of decanoic acid, conditions under which the coacervate (around 340 microL) formed in situ and instantaneously. The overall sample treatment, which included extraction and centrifugation, took about 25-30 min, and several samples could be simultaneously treated using conventional lab equipment. No clean-up or solvent evaporation were required. Extraction efficiencies mainly depended on the decanoic acid and THF concentration in the aqueous solution and were not affected by the pH or the temperature in the ranges studied (1-4 and 20-60 degrees C, respectively). Recoveries in samples ranged between about 81 and 96%. The precision of the method, expressed as relative standard deviation, was about 3% and the quantitation limit was around 9 ng g(-1), which was far below the current specific migration limit (SML) set for BPA by the EU Commission (600 ng g(-1)). The method was successfully applied to the determination of BPA in the solid content of canned fruit salad, peaches in syrup, mango slices, red peppers, sweetcorn, green beans and peas. BPA was present at concentrations in the range from 7.8 to 24.4 ng g(-1) in canned fruits and from 55 to 103 ng g(-1) in canned vegetables.     

Determination of phthalates in wine by headspace solid-phase microextraction followed by gas chromatography-mass spectrometry: fibre comparison and selection

This paper describes the development of a headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) method for determining phthalates in wine. The HS-SPME conditions were thoroughly studied: first, the performance of six fibres at three temperature values and two sample volumes was surveyed by means of a 6 x 3 x 2 multi-factor categorical experimental design. From this study, three fibres - carbowax-divinylbenzene (CW-DVB), polyacrylate (PA) and polydimethylsiloxane-divinylbenzene (PDMS-DVB) - were selected. Then, temperature, sample volume and sodium chloride concentration were optimised using a central composite design and the overall desirability function for each fibre. The optimal values were 70 degrees C, a NaCl concentration of 2.6, 3.6 and 5.5M for PA, CW-DVB and PDMS-DVB fibres, respectively, and sample volumes of 4.0, 3.5 and 3.0 mL. Next, the performance characteristics of the three fibres were obtained and compared. PDMS-DVB fibre showed the best repeatability values followed by CW-DVB. PA fibre was not suitable for diethylhexylphthalate extraction and showed poor repeatability for the heavier phthalates, and was therefore discarded. Finally, the performance of CW-DVB and PDMS-DVB fibres was checked for red, white and rosé wines.     

Comparison of different immunoaffinity clean-up procedures for high-performance liquid chromatographic analysis of ochratoxin A in wines

Three immunoaffinity clean-up procedures to analyse ochratoxin A (OTA) in wines were compared. The direct wine clean-up with Ochraprep and OchraTest columns gave equivalent results in terms of recovery and precision if compared with the reference procedure involving a preliminary extraction of OTA with chloroform. OTA quantification limit in wine ranged from 0.020 to 0.045 microg/l. The 'on-flow' OTA emission spectrum (excitation 333 nm) showed a maximum at 460 nm and could be used to confirm the quantitative results. The analysis of 11 red and white wines gave no significant quantitative differences between the three clean-up techniques.     

An electrochemical immunosensor for ochratoxin A determination in wines based on a monoclonal antibody and paramagnetic microbeads

We report a direct competitive immunosensor for the rapid determination of ochratoxin A (OTA) in wine samples. Magnetic beads (1 ± 0.5 μm diameter) covered with streptavidin were functionalized with a monoclonal antibody against OTA, and then left to incubate in a solution of tracer (ochratoxin conjugated to the enzyme peroxidase) and a range of OTA concentrations (10(-4) to 1,000 ng mL(-1)). After washing and separation steps helped with a magnetic field, a volume of the dispersion was put on screen-printed electrodes under a magnet, and after adding the substrate the p-benzoquinone generated enzymatically was detected by differential-pulse voltammetry. Wine samples (2 mL) were easily prepared simply by adjusting to pH = 7.5 with diluted NaOH and by adding polyvinylpyrrolidone for complexing polyphenols, without any other clean-up or preconcentration steps. The limit of detection for detecting OTA in wines was of 0.11 ± 0.01 ng L(-1), well below the permitted content of the mycotoxin by the European Union (<2 ng mL(-1)). Spiked wines were subjected to immunosensor calibrations to study the matrix effects. OTA concentrations measured with the immunosensor were compared with those obtained by high-performance liquid chromatography coupled to fluorescence detection (AOAC official method 2001.01). The OTA levels from two red wines of "Campo de Borja", Spain, ranged from about 0.027 to 0.033 ng mL(-1) of OTA.     

Concentration of ochratoxin A in wines from supermarkets and stores of Valencian Community (Spain)

Ochratoxin A (OTA) is a mycotoxin produced by fungi species belonging to the genera Aspergillus and Penicillium being isolated in alcoholic beverages. The aim of this work is developed and applied a procedure for the analysis of OTA in wines. An analytical method based on immunoaffinity column (IAC) for clean-up, liquid chromatography with fluorescence detection (LC-FD), and LC-FD after of OTA methylation was used to determine the occurrence of OTA in wines. Recoveries of this mycotoxin spiked to red wines at 0.5 ng/ml level were >90% with an average of relative standards deviations of 4%. Furthermore, 116 wine samples from designation of origin (DO) and three samples from food stores of Valencian Community (Spain) were examined for the occurrence of OTA being the levels of this mycotoxin ranged from <0.01 to 0.76 ng/ml. Finally, the estimated daily intake of OTA in this study was 0.15 ng/kg bw per day.     

Determination of phenolic compounds in wines by novel matrix solid-phase dispersion extraction and gas chromatography/mass spectrometry

A novel matrix solid-phase dispersion (MSPD) extraction method was developed to extract simultaneously 23 phenolic compounds from wine samples prior to determination by gas chromatography with mass spectrometric detection in the selected ion monitoring mode. Different parameters of the MSPD technique such as dispersant solid-phase, eluting solvent, and sample ionic strength and pH were optimized. The optimized MSPD procedure requires a small volume of wine (1 mL), commercial silica gel (1.5 g) as dispersant solid-phase and a small volume of ethyl acetate (5 mL) as eluting solvent. Under these conditions, the extraction of the studied compounds was almost complete (mean values of recoveries between 87 and 109%) in a short time (15 min). Moreover, satisfactory standard deviations of repeatability (RSD<9% in most cases), linear regression coefficients (r(2)>0.993) and detection limits (<8 microg/L) confirm the usefulness of the methodology for routine monitoring of the concentration of individual phenolic antioxidants in wines. Application was illustrated by analysis of different wine samples.     

Fluorescence polarization immunoassay for rapid screening of ochratoxin A in red wine

A fluorescence polarization (FP) immunoassay, based on a monoclonal antibody and an ochratoxin A (OTA)-fluorescein tracer, has been developed for rapid screening of OTA in red wine. Wine samples were diluted with methanol and passed through aminopropyl solid-phase extraction columns prior to the FP assay. Average recoveries from samples spiked with OTA at levels of 2.0 and 5.0 ng/mL were 79% with RDS of 11% (n = 6). The limit of detection of the FP immunoassay was 0.7 ng/mL OTA, and the whole analysis was performed in less than 10 min. The assay was tested on 154 red wine samples (naturally contaminated or spiked at level ranging from 0.1 to 5.0 ng/mL) and compared with an high-performance liquid chromatography/immunoaffinity column clean-up method, showing a good correlation (r = 0.9222). Their compliance with the European regulation (2.0 ng/mL OTA maximum permitted level) was correctly assessed for 70% of the analyzed samples of red wine, whereas confirmatory analyses were required for the remaining ones with OTA levels close to the regulatory limit. No false-negative or positive results were observed using the FP immunoassay. The proposed FP assay is a useful screening method for OTA in red wines, when high throughput is required, that could also be used for white and rosé wines, which are known to contain less interfering compounds such as polyphenols.     

Validation of two analytical methods for the determination of ochratoxin A by reversed-phased high-performance liquid chromatography coupled to fluorescence detection in musts and sweet wines from Andalusia

Some Spanish sweet wines are made from raisins, grapes dried by direct exposure to the sun after picking. This drying process can encourage ochratoxin A (OTA) formation. OTA is a mycotoxin formed by several fungi. It has been linked to nephropathy in humans, and may have a long half-life in humans. The aim of this study is to develop and to apply two procedures for the analysis of OTA in grape musts (during the raisining process) and sweet wines, respectively. Reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to fluorescence detection (FLD) was employed in both analytical methods. In grape must, the method involves the direct injection of the sample in a HPLC-FLD system without any kind of prior clean-up procedure. The complexity of the sweet wine samples requires a solid-phase extraction (SPE) clean-up on a C18 column which enables the OTA to be isolated from the matrix. The methods used were statistically validated. The validation also included the comparison of the slopes of the curve obtained with standards and the regression curves obtained by the addition of a standard. Two different studies of standard additions were conducted. One method was validated without sample preparation and it was applied to must samples. The other method was validated with SPE extraction and it was applied to sweet wine samples. Recovery was always better than 89.69%. The limit of detection (S/N = 3) and limit of quantification (S/N = 10) were established at 0.22 and 0.77 μg l⁻¹, respectively. In general, the analytical data obtained provided good results at the sub-μg l⁻¹ concentration level.     

Supramolecular solvent-based microextraction of ochratoxin A in raw wheat prior to liquid chromatography-fluorescence determination

A supramolecular solvent made up of reverse micelles of decanoic acid, dispersed in a continuous phase of THF:water, was proposed for the simple, fast and efficient microextraction of OTA in wheat prior to liquid chromatography-fluorescence determination. The method involved the stirring of 300 mg-wheat subsamples (particle size 50 μm) and 350 μL of supramolecular solvent for 15 min, subsequent centrifugation for 15 min and the direct quantitation of OTA in the extract, previous 5.7-fold dilution with ethanol/water/acetic acid (49.5/49.5/1), against solvent-based calibration curves. No clean-up of the extracts or solvent evaporation was needed. Interactions between the supramolecular solvent and major matrix components in the wheat (i.e. carbohydrates, lipids and proteins) were investigated. The reverse micelles in the extractant induced gluten flocculation but only in the coacervation region of lower analytical interest (i.e. at percentages of THF above 11%). The quantitation of OTA was interference-free. Representativity of the 300 mg-wheat subsamples was proved by analysing a reference material. OTA recoveries in wheat ranged between 84% and 95% and the precision of the method, expressed as relative standard deviation, was 2%. The quantitation limit of the method was 1.5 μg kg⁻¹ and was below the threshold limit established for OTA in raw cereals by EU directives (5.0 μg kg⁻¹). The method developed was validated by using a certified reference material and it was successfully applied to the determination of OTA in different wheat varieties from crops harvested in the South of Spain. OTA was not detected in any of the analysed samples. This method allows quick and simple microextraction of OTA with minimal solvent consumption, while delivering accurate and precise data.     

Direct sample introduction of wines in graphite furnace atomic absorption spectrometry for the simultaneous determination of arsenic, cadmium, copper and lead content

A multi-element graphite furnace atomic absorption spectrometry (GFAAS) method was elaborated for the simultaneous determination of As, Cd, Cu, and Pb in wine samples of various sugar contents using the transversally heated graphite atomizer (THGA) with end-capped tubes and integrated graphite platforms (IGPs). For comparative GFAAS analyses, direct injection (i.e., dispensing the sample onto the IGP) and digestion-based (i.e., adding oxidizing agents, such as HNO(3) and/or H(2)O(2) to the sample solutions) methods were optimized with the application of chemical modifiers. The mixture of 5 microg Pd (applied as nitrate) plus 3 microg Mg(NO(3))(2) chemical modifier was proven to be optimal for the present set of analytes and matrix, it allowing the optimal 600 degrees C pyrolysis and 2200 degrees C atomization temperatures, respectively. The IGP of the THGA was pre-heated at 70 degrees C to prevent the sputtering and/or foaming of sample solutions with a high organic content, dispensed together with the modifier solution, which method also improved the reproducibility of the determinations. With the digestion-based method, the recovery ranged between 87 and 122%, while with the direct injection method it was between 96 and 102% for Cd, Cu, and Pb, whereas a lower, compromise recovery of 45-85% was realized for As. The detection limits (LODs) were found to be 5.0, 0.03, 1.2, and 0.8 microg l(-1) for As, Cd, Cu, and Pb, respectively. The characteristic mass (m(0)) data were 24 pg As, 1.3 pg Cd, 13 pg Cu, and 35 pg Pb. The upper limits of the linear calibration range were 100, 2, 100, and 200 microg l(-1) for As, Cd, Cu, and Pb, respectively. The precisions were not worse than 4.8, 3.1, 3.7, and 2.3% for As, Cd, Cu, and Pb, respectively. For arsenic, a higher amount of the modifier (e.g., 20 microg Pd plus 12 microg Mg(NO(3))(2)) could be recommended to overcome the interference from the presence of sulphate and phosphate in wines. Although this method increased the sensitivity for As (m(0)=20 pg), it also enhanced the background noise, thus only a slight improvement in the LOD of As (3.9 microg l(-1)) was realized. For the 35 red and white wine samples studied, the highest metal contents were observed for Cu ranging from 20 to 640 microg l(-1) (average: 148 microg l(-1)), followed by Pb from 6 to 90 microg l(-1) (average: 32.3 microg l(-1)), and Cd from 0.05 to 16.5 microg l(-1) (average: 1.06 microg l(-1)), whereas the As content was below the LOD. This wide fluctuation in the trace metal content could be associated with the origin of wines from various regions (i.e., different trace metal level and/or quality of soil, and/or anthropogenic impact), and with diverse materials (e.g., additives and containers) involved in the wine production processes. The Cu content of wine samples was significantly correlated with Pb, whereas its weak anti-correlation was found with Cd. Interestingly, the level of Pb was anti-correlated with the year of production of the wines. This is likely due to the gradual decrease in the Pb content of soils of vineyards by time, which certainly causes less Pb-uptake of the grape plant, thus a decrease in the Pb content of wines as well.