Jesús Sanz-Asensio

Comparison of four extraction methods for the determination of fungicide residues in grapes through gas chromatography-mass spectrometry.

Four different methods for simultaneous extraction of vinclozolin, dichlofluanid, penconazole, captan, quinoxyfen, fluquinconazol, boscalid and pyraclostrobin from grapes were optimized and further tested. Microwave assisted extraction (MAE), matrix solid-phase dispersion (MSPD), solid-liquid extraction (SLE) and QuEChERS were compared in terms of their limits of detection and quantification and recoveries. For MAE, MSPD and ethyl acetate extraction, the optimal conditions were optimized by using experimental designs. The analysis was performed using gas chromatography-mass spectrometry in the selected ion monitoring mode (GC-MS, SIM). The proposed methods showed good sensitivity, limits of quantification were lower than MRLs and precision (expressed as relative standard deviation) ranged from 2.9 to 11.1%. The recoveries obtained from MAE, MSPD, SLE and QuEChERS were in the range 78-100%, 66-102%, 58-88% and 68-96%, respectively. In addition, the four methods were compared in two ways: by means of calibration curves obtained with 10 fortified samples in the studied range of concentrations and by the application of statistical tests such as Levenes test (to study variance homogeneity), ANOVA and Tukeys test (in case of Levenes test was satisfactory) for the assessment of the information obtained in the analysis of real samples. Both ways of comparison led to the same results: no differences between the four methods for the extraction of vinclozolin, dichofluanid, quinoxyfen, fluquinconazol and pyraclostrobin were found. However, there were differences for the analysis of captan, boscalid and penconazole.

Study of photodegradation of the pesticide ethiofencarb in aqueous and non-aqueous media, by gas chromatography–mass spectrometry

A comparative photodegradation kinetic study of ethiofencarb [2-ethylthiomethyl(phenyl)-N-methylcarbamate] in aqueous and non-aqueous media (hexane and methanol), is carried out. After irradiation, the aqueous samples are extracted with isobutyl methyl ketone. Ethiofencarb and its metabolites are analyzed by GC in combination with nitrogen–phosphorus detection and MS, respectively. The degradation kinetics depend on the solvent polarity; the quickest pesticide transformation is in the aqueous medium and the slowest in hexane. The photoproducts are also dependent on the solvents. In the case of the aqueous solution, photocleavage of the carbon–sulphur bond gives 2-(methyl)phenyl-N-methylcarbamate as the main product, while methanol and hexane solutions show different photoproducts.

Kinetic study of the degradation of ethiofencarb in aqueous solutions

A kinetic study of the hydrolysis of ethiofencarb (α-ethylthio-o-tolyl methylcarbamate) in pure water and in aqueous solutions at pH 2, 6, 9 and 12 and at three different temperatures (4, 20 and 50(±1)°C) has been carried out using a gas chromatographic nitrogen-phosphorus detection method. The values of the first-order rate constants (k) for the degradation reaction were calculated. The values for k were found to be dependent on pH and temperature. No acid hydrolysis was observed in any case. Complete degradation of ethiofencarb was observed at pH 12 at all three temperatures; it was practically instantaneous at room temperature. Ethiofencarb was also completely degraded at pH 9 at 20 and 50°C, while in pure water (pH 6) degradation took place at 50°C but not at 20°C. Ethiofencarb was not degraded in pure water at lower temperatures and, due to the reversible nature of the reaction, at equilibrium about 80% of the pesticide remained undegraded at room temperature.

Behaviour of acephate and its metabolite methamidophos in apple samples

SummaryA study of the decay of acephate in apple samples was carried out, including penetration studies and the transformation of acephate in to its main metabolite, methamidophos. Sample treatment involved extraction with ethyl acetate and determination by gas chromatography with nitrogen—phosphorus detection (GC-NPD). Three different parts of the fruit were studied separately: apple surface, peel and pulp. Recoveries were measured at three spiked levels, ranging from 0.050 to 0.504 μg g−1 for acephate and 0.049 to 0.492 μg g−1 for methamidophos. Mean acephate recoveries were 93.0 to 115.5% from peel and 99.2 to 110.2% from pulp, while methamidophos recoveries were 77.2 to 104.2% and 77.5 to 98.6% from peel and pulp, respectively (n=6). Results showed that acephate penetrates into the fruit, where it is transformed to methamidophos. This transformation was not seen on the external apple surface.

Effect of Cluster Thinning and Prohexadione Calcium Applications on Phenolic Composition and Sensory Properties of Red Wines

The overall objective of this study was to investigate the effect of manual cluster thinning (CT) and the application of the growth regulator Prohexadione calcium (ProCa) on the phenolic composition and the sensory profile of Tempranillo and Grenache wines produced from treated vines in La Rioja (Spain). ProCa was applied at preblooming and CT was carried out at veraison in two consecutive years. Different physicochemical parameters and analyses of phenolic compounds were carried out in control, CT and ProCa grapes and wines and wine sensory was performed. Thinning treatments decreased crop yield, besides ProCa application reduced berry size, and berry weight. Color and phenolic composition of Grenache and Tempranillo wines in general were affected by thinning treatments, with an increase in anthocyanin, flavanol and flavonol concentrations. In sensory analysis, wines obtained from thinned vines presented higher values for several aromatic (e.g., white and yellow fruits, fresh flowers) and taste attributes (i.e., astringency, bitternes, persistence). CT and ProCa treatments resulted in an improvement in wine quality. In general, similar results in phenolic composition, sensory properties and quality of wines were obtained by manual and chemical cluster thinning. ProCa as a growth regulator may be an option for a quality vitiviniculture.

Validation of a microwave-assisted extraction gas chromatography detection method for the determination of fungicides in grapes

A microwave assisted extraction (MAE) method was developed for the simultaneous extraction of eight pesticides, vinclozolin, dichlofluanid, penconazole, captan, quinoxyfen, fluquinconazole, boscalid and pyraclostrobin, in grapes. The different parameters affecting the MAE method, such as temperature, extraction time and nature and volume of the extraction solvent, were evaluated. The experimental procedure was optimized using a multivariate statistical approach and the final analyses were carried out by gas chromatography-mass spectrometry (GC-MS). These eight fungicides could be efficiently extracted from 2.0 g of grapes with 10 mL of hexane/acetone (1 : 1, v/v) mixture at 105 °C in 10 minutes with microwave power set at 100% (600 W). Using the optimized conditions, the method was validated in terms of accuracy and precision. A matrix effect study was also carried out using different grape samples. Recoveries from spiked grape samples ranged from 82 to 107% and relative standard deviations were less than 10%. Detection and quantification limits ranged from 0.7 to 1.7 μg kg−1 and 2.2 to 5.8 μg kg−1, respectively, which were lower than the maximum residue limits of the pesticides in grape samples established by the European Union. Uncertainty assessment of measurement obtained by applying the proposed analytical method was estimated according to EURACHEM/CITAC guide. The expanded uncertainties were below 26% and 7% at two concentration levels, 10 and 50 μg kg−1, respectively.

Dissipation of Fungicide Residues during Winemaking and Their Effects on Fermentation and the Volatile Composition of Wines

The effects of four fungicides commonly used for the control of fungal diseases in vines and grapes in the course of winemaking were tested. The concentration of fungicide residues was monitored throughout the process to establish their kinetics of dissipation. In all cases the percentages of dissipation were >68%, which shows the detoxificant effect of the winemaking process. On the other hand, the effect of the fungicide residues on the aroma composition of Tempranillo red wines was tested. To evaluate possible modifications on the aroma profile of wines, seven odorant series (ripe fruits, fresh fruits, lactic, floral, spicy, vinous, and herbaceous) were built from the odor activity values (OAVs) obtained for each volatile compound. Ripe fruits and fresh fruits were the major aromatic attributes in all Tempranillo red wines. These two odorant series registered the highest variations in their total OAVs with respect to the control wine, especially with the application of boscalid + kresoxim-methyl into vines, leading to a decrease in the ripe fruit and fresh fruit nuances of the resulting wines. Moreover, when the effect of these fungicides on the aroma of Tempranillo red wines was compared throughout two years (2012 and 2013), wines elaborated from grapes treated in the field with boscalid + kresoxim-methyl in 2013 displayed the highest variation in aroma profile with respect to control wine.

Determination of fungicide pyrimethanil in grapes, must, fermenting must and wine

BACKGROUND This study determined the evolution of pyrimethanil, a fungicide commonly used to control Botrytis cinerea, throughout the winemaking process in grapes, must, fermenting must and wine. Tempranillo grapevines were treated with pyrimethanil according to both good Agricultural Practices (GAP) and Critical Agricultural Practices (CAP). Fermentation was carried out in an experimental winery. Grape analysis was based on an ethyl acetate extraction method. Samples from fermentation were analysed by solid phase extraction. The determination was carried out by gas chromatography with nitrogen-phosphorus detection (GC-NPD) and additionally confirmed by gas chromatography/mass spectrometry (GC/MS). RESULTS Pyrimethanil residues were at least ten times greater in grapes treated 7 days before harvest than in those treated respecting the safety period (21 days). The amount of pyrimethanil in grapes treated under GAP was below the maximum residue limit (5 mg kg(-1)). The level of pyrimethanil decreased during fermentation in both treatments. In the fermentation of grapes treated according to CAP, the pyrimethanil concentration was reduced by over 50% on the first day and then remained constant until the end of the fermentation process. For grapes treated in compliance with GAP, the amount of pyrimethanil decreased to a level below the limit of detection in the bottled wine. CONCLUSION The described methods for grapes, must, fermenting must and wine gave good recoveries, linearity, precision and accuracy. They were also highly sensitive in avoiding matrix effects. Pyrimethanil residues found in treated grapes were higher in skin than in pulp. The amount of pyrimethanil decreased during fermentation by degradation and/or adsorption. For grapes treated according to GAP, residues disappeared in the final bottled wine. The decrease observed in the final bottled wine may be caused by diverse oenological practices and technologies such us malolactic fermentation, racking and settling.

Analysis of Pyrimethanil, Metalaxyl, Dichlofluanid, and Penconazol in Must and Wine from Red Grapes by Solid-Phase Extraction and Gas Chromatography

Abstract This article describes a method for the simultaneous analysis of four fungicides—pyrimethanil, penconazol, metalaxyl, and dichlofluanid—in must and wine by solid-phase extraction. Two cartridges (octadecylsilane and polystyrene-divinylbenzene) were evaluated with different elution solvents. The most efficient solid phase was octadecylsilane together with ethyl acetate as elution solvent. Determination was performed by gas chromatography with a nitrogen–phosphorus detector, using metribuzin as internal quantification standard. To reduce quantitative errors from matrix effects, matrix-matched calibration was used. The linear regression coefficients were at least 0.9990. The quality parameters of the proposed method presented good recoveries from spiked must and wine, ranging from 92.3 to 105.5%, and good precision, indicated by relative standard deviations (RSDs) of less than 10%. The limits of quantification varied from 0.27 µg L−1 for penconazol in wine to 9.34 µg L−1 for metalaxyl in wine. An additional confirmation method, gas chromatography with mass spectrometry detection, was used.