Degradation studies of dimethachlor in soils and water by UHPLC-HRMS: putative elucidation of unknown metabolites.

Degradation of dimethachlor was evaluated in soils and water, detecting metabolites in incurred samples. Putative elucidation was performed using HRMS and software tools, detecting new possible metabolites of dimethachlor.

BACKGROUND: The analytical control of the presence of dimethachlor and its metabolites in environmental samples, such as water and soils, is a main concern. Degradation of this pesticide has been evaluated in two types of soils and two different water conditions at two concentration levels. For that purpose, a new liquid chromatography-mass spectrometry method has been developed and putative identification of new metabolites has been performed. RESULTS: An analytical method based on ultra-high-performance liquid chromatography coupled to Orbitrap mass spectrometry (UHPLC-Orbitrap-MS) was developed to monitor the degradation of dimethachlor in environmental samples (water and soils). The degradation of dimethachlor in soils and groundwater samples has been monitored from 1 to 110 days after application of a plant protection product at two doses. Concentration of the parent compound slowly decreased in both matrices. DT50 values ranged from 40 to 70 days. Some metabolites were detected in the commercial product and in the samples one day after the application of the plant protection product. In addition, three new metabolites were putatively identified during dimethachlor degradation by untargeted analysis. CONCLUSION: In this study, the degradation of dimethachlor into its metabolites has been studied in soils and water, using a UHPLC-Orbitrap-MS validated method. A putative elucidation of new metabolites of dimethachlor has been carried out applying HRMS and software tools. Degradation results allowed for understanding the behavior of dimethachlor in soils and water, and provided information regarding the possible risk of this pesticide and its metabolites to the ecosystem.

Determination of quaternary ammonium compounds in oranges and cucumbers using QuEChERS extraction and ultra-performance liquid chromatography/tandem mass spectrometry.

A simple and fast method has been developed for determining relevant quaternary ammonium compounds in cucumber and orange samples. The target compounds were benzoalkonium chloride (BAC-10, BAC-12, BAC-14, and BAC-16), didecyldimethylammonium chloride, and benzethonium chloride, all frequently used biocides in the agrifood industry. An extraction based on the buffered Quick, Easy, Cheap, Effective, Rugged, and Safe method and determination by ultra-performance LC/MS/MS that eluted the biocides in less than 5 min were used. The method was fully validated and implemented in a UNE-EN-ISO/IEC 17025 accredited laboratory for its application to the analysis of real samples. Performance characteristics of the method are reported, including an estimation of measurement uncertainty. Calibration curves were set between 0.01 and 0.150 mg/kg, LOD values were always between 0.4 and 1.0 microg/kg, LOQ values were in the range 1-4 microg/kg, recovery was between 81 and 115%, intraday and interday precision were always lower than 17% (expressed as RSD), and expanded uncertainty was always lower than 40%. The validation was accomplished for the two studied matrixes at spiking concentrations of 0.011 and 0.050 mg/kg. The method has been applied to the analysis of 30 cucumber and orange samples that were found to contain concentrations of BAC-12 that ranged between 0.015 and 0.210 mg/kg and of BAC-14 between 0.018 and 0.081 mg/kg.

QuEChERS approach for the determination of biopesticides in organic and nonorganic vegetables and fruits by ultra-performance liquid chromatography/tandem mass spectrometry.

A method for the determination of 14 biopesticides and piperonyl butoxide (PBO), often applied in organic farming, in vegetables and fruits has been developed. Extraction was performed using the Quick, Easy, Cheap, Effective, Rugged, and Safe method, and the determination was carried out by ultra-performance LC/MS/MS in 9 min. Several different food commodities were evaluated as representative matrixes, namely, cucumber, tomato, pepper (high water content), and strawberry and orange (high acid and water content). Biopesticides were quantified using matrix-matched calibration, and the optimized method was validated at three concentration levels, i.e., 10, 50, and 100 microg/kg, for all the compounds, yielding recoveries in the range 70-112%, 71-112%, and 70-109%, respectively, with RSDs <28%. LOQs were <3 microg/kg for all biopesticides. The validated procedure was applied to the analysis of vegetables and fruits, including products from organic farming; PBO was detected in one sample at 8 microg/kg.

Systematic study of the contamination of wastewater treatment plant effluents by organic priority compounds in Almeria province (SE Spain).

The occurrence of priority organic pollutants in wastewater (WW) effluents was evaluated in a semi-arid area, characterized by a high agricultural and tourism activity, as Almeria province (Southeastern Spain). Twelve wastewater treatment plants (WWTPs) were sampled in three campaigns during 2011, obtaining a total of 33 WW samples, monitoring 226 compounds, including pesticides, polycyclic aromatic hydrocarbons (PAHs), phenolic compounds and volatile organic compounds (VOCs). Certain banned organochlorine pesticides such as aldrin, pentachlorobenzene, o,p'-DDD and endosulfan lactone were found, and the most frequently detected pesticides were herbicides (diuron, triazines). PAHs and VOCs were also detected, noting that some of these pollutants were ubiquitous. Regarding phenolic compounds, 4-tertoctylphenol was found in all the WW samples at high concentration levels (up to 89.7 µg/L). Furthermore, it was observed that WW effluent samples were less contaminated in the second and third sampling periods, which corresponded to dry season. This evaluation revealed that despite the WW was treated in the WWTP, organic contaminants are still being detected in WW effluents and therefore they are released into the environment. Finally the risk of environmental threat due to the presence of some compounds in WWTP effluents, especially concerning 4-tertoctylphenol must be indicated.

Determination of pesticides in water samples by solid phase extraction and gas chromatography tandem mass spectrometry.

A multiresidue method for the determination of more than 80 pesticides in water has been developed and validated. The proposed method is based on SPE followed by GC coupled to MS/MS. Different variables affecting SPE procedure, such as cartridges, sample volume and solvents were studied, and mass spectrometric conditions were optimised in order to increase selectivity and sensitivity. Calibration curves were linear over the range of 0.03-0.5 microg/L. Recoveries were in the range of 70-110% and repeatability was below 20% for the lowest calibration point. LODs ranged from 0.001 to 0.025 microg/L and LOQs from 0.003 to 0.076 microg/L. Finally, the method was successfully applied to the analysis of water samples from southeast of Spain.

Determination of polycyclic aromatic hydrocarbons in olive oil by a completely automated headspace technique coupled to gas chromatography-mass spectrometry.

A new and completely automated method for the determination of ten relevant polycyclic aromatic hydrocarbons (PAHs) in olive oil is proposed using an extraction by the headspace (HS) technique. Quantification and confirmation steps are carried out by gas chromatography-mass spectrometry (GC-MS) combining simultaneously selected-ion monitoring (SIM) and tandem mass spectrometry (MS/MS). This combination offers on one hand an increased sensitivity and on the other hand, selective and reliable qualitative information. Sample pretreatment or clean-up are not necessary because the olive oil sample is put directly into an HS vial, automatically processed by HS and introduced into the GC-MS instrument for analysis. Because of its high selectivity and sensitivity, a triple-quadrupole (QqQ) detector coupled with the gas chromatograph allows us to limit handling. Each sample is completely processed in approximately 63 min (45 min for HS isolation and 18 min for GC-MS determination), a reduced time compared with previously published methods. The chemical and instrumental variables were preliminarily optimized using uncontaminated olive oil samples spiked with 25 microg kg(-1) of each target compound. The final method was validated to ensure the quality of the results. The precision was satisfactory, with relative standard deviation (RSD) values in the range 3-9%. Recovery rates ranged from 96 to 99%. Limits of detection (LOD) were calculated as 0.02-0.06 microg kg(-1) and the limits of quantification (LOQ) were obtained as 0.07-0.26 microg kg(-1). It must be mentioned that the LOD and LOQ are much lower than the maximum levels established by the European Union (EU) in oils and fats intended for direct human consumption or for use as an ingredient in foods, which are set at 2 microg kg(-1). All the figures of merit are completely in accordance with the latest EU legislation. This fact makes it possible to consider the proposed method as a useful tool for the control of PAHs in olive oils.

Potentiality of gas chromatography-triple quadrupole mass spectrometry in vanguard and rearguard methods of pesticide residues in vegetables.

A new analytical strategy for the screening and confirmation/quantification of multiclass pesticide residues in vegetables has been established and validated. No complicated sample preparation was needed, but only a simple and rapid extraction using ethyl acetate and sodium sulfate, which required no cleanup. The approach is based on the use of the triple quadrupole (QqQ) mass spectrometry (MS) as detection system in gas chromatography (GC). In a first step, a GC-QqQ-MS screening method, which monitors only one MS/MS transition by compound, allows the identification of approximately 130 pesticides in 11.6 min. In this way, the differentiation between negative and potentially nonnegative samples is carried out. In the second step, the nonnegative samples are reanalyzed by the GC-QqQ-MS confirmation/quantification method, which monitors two or three MS/MS transitions by compound. Confirmation of pesticides was based on the comparison of intensity ratios for the main ions in samples with those obtained on the same day from the standard in a matrix containing the pesticides at a preestablished concentration level. Quantification of the identified and confirmed pesticides was based on the addition standard method, which avoids matrix effect. The proposed analytical strategy allowed a reliable identification and confirmation of the target pesticides at trace levels, reducing analysis time and increasing sample throughput in routine analytical laboratories.

A study of the disappearance of pesticides during composting using a gas chromatography-tandem mass spectrometry technique.

A simple method for multi-residue analysis of pesticides by low-pressure gas chromatography-tandem mass spectrometry (LP-GC-MS-MS) has been validated in compost samples. The pesticide residues were extracted from the lyophilised samples with organic solvent by stirring. No sample clean-up was required prior to the analysis. The method was applied to determine the fate of two organochlorine pesticides (lindane and endosulfan) and two organophosphates (malathion and chlorpyrifos-methyl) during the composting process in the reactor of a pilot plant. Malathion, chlorpyrifos-methyl and lindane residues almost fully disappeared after 8 days of maturation in the reactor, while endosulfan residues were only partially degraded. Alongside this, a study of natural disappearance of the pesticides was carried out in the laboratory and a characteristic time profile was traced for each pesticide class.

Chemiluminescence determination of carbofuran at trace levels in lettuce and waters by flow-injection analysis.

A simple, fast chemiluminescence (CL) flow-injection (FI) method based on the reaction of luminol with KMnO(4) in alkaline medium has been described for the direct determination of carbofuran. The method is based on the enhancing effect in the emission light from the oxidation of luminol produced in presence of carbofuran. The optimisation of instrumental and chemical variables influencing the CL response of the method has been carried out by applying experimental design, using the proposed flow-injection manifold. Under the optimal conditions, the CL intensity was linear for a carbofuran concentration over the range of 0.06-0.5mugml(-1), with a detection limit of 0.02mugml(-1). The method has been successfully applied to the determination of carbofuran residues in spiked water and lettuce samples.

Assessment of metal contamination in Doñana National Park (Spain) using crayfish (Procamburus clarkii).

Water quality assessment in the Aznalcollar area was attempted using multivariate methods based on heavy metal concentrations in red swamp crayfish (Procamburus clarkii). Trace levels of four heavy metals, copper (Cu), zinc (Zn), cadmium (Cd) and lead (Pb), were detected in crayfish from eleven different stations. Principal component analysis (PCA) highlighted a gradient of contamination between the sampling stations. Cluster analysis (CA) distinguished three groups of stations. Discriminant analysis also differentiated three groups. The group centroids of the first discriminant function were used to devise an index that varies according to the source of the crayfish. These standardized values are proposed for use as a water quality index. The ability of this index to successfully predict environmental quality was proved with random samples.

Assessment of total effective xenoestrogen burden in adipose tissue and identification of chemicals responsible for the combined estrogenic effect.

Test systems to screen for estrogenicity and appropriate biomarkers of human exposure are required for epidemiological studies of endocrine disruption. We addressed these issues by developing and standardising a method to assess the total estrogenic xenobiotic burden in human adipose tissue. In this study, which is the continuation of a previous work, we have improved the protocol for extensive fractionation of a higher number of tissue samples in order to investigate bioaccumulated xenoestrogens that are candidates for estrogenicity and to assess their combined estrogenic effect. This was achieved by extensive HPLC separation of xenoestrogens from endogenous hormones followed by testing of individual fractions in the E-Screen test for estrogenicity. Organochlorine pesticides, PCBs and halogenated bisphenols and alkylphenols were collected in the most lipophilic fractions, followed by progestins, androgens and estradiol esters, and then by steroidal estrogens; phyto- and myco-estrogens were collected around the end of the run. These results were confirmed by exhaustive chemical analysis. In 458 human adipose tissue samples, the total effective xenoestrogen burden was positive in 75% of samples in the pooled fraction that contained organohalogenated xenoestrogens (mean 515.3 pM Eeq/g lipid; range 0-14.5 nM) and in 82% of samples in the pooled fraction where natural estrogens eluted (mean 696.6 pM Eeq/g lipid; range 0-12.9 nM). Organochlorine pesticides emerged as candidate chemicals for the estrogenicity of the first pooled fraction, because DDT and derivatives were present in 98.3% of the samples. However, no correlation was found between the concentration of any single chemical and the estrogenicity determined in the bioassay. There may be several reasons for this lack of concordance: (i) the estrogenic effects depicted in the E-Screen bioassay are a consequence of the combined effect of several organohalogens or (ii) the proliferative effect is due to other chemicals not measured. Because additive, synergistic or antagonistic mechanisms may account for the final effect observed in the pooled fractions, the approach proposed in this work is more appropriate for exposure assessment in epidemiological studies than the determination of individual chemicals in human samples.

Benzoylphenylurea residues in peppers and zucchinis grown in greenhouses: determination of decline times and pre-harvest intervals by modelling.

Residue levels and degradation rates of five benzoylphenylurea insecticides were studied in zucchinis and peppers grown in experimental greenhouses in Almería (Spain). Benzoylphenylurea residues were analyzed by HPLC using on-line post-elution photoirradiation with fluorescence detection. Mathematically defined decline curves were established by determining optimal relationships between benzoylphenylurea residues and time, using different models. The models that best fitted the experimental data were those of first-order for diflubenzuron, triflumuron, hexaflumuron and flufenoxuron in zucchini and RF first-order models for the five insecticides in peppers and for lufenuron in zucchini. Half-life times for the residues on the two vegetables were estimated from the optimal models. In order to guarantee safe consumption of the two vegetables, we have estimated suitable pre-harvest intervals complying with the maximum residue levels established by the Spanish Government. In all cases, such pre-harvest intervals were shorter than those specified by the manufacturers of commercial formulates. Experimental data for the five insecticides in peppers and for lufenuron in zucchini were also fitted to a first-order model. Even though this function was legitimized statistically, estimations of decline times (T/2) and pre-harvest intervals were quite different from those provided by the optimal model.

Selective extraction and determination of multiclass pesticide residues in post-harvest French beans by low-pressure gas chromatography/tandem mass spectrometry.

A selective and reliable extraction procedure was developed and validated to determine multiclass pesticide residues in lyophilized agricultural plants by low-pressure gas chromatography coupled with tandem mass spectrometry. The method is based on a rapid and simple extraction of the lyophilized sample with dichloromethane. The results were compared with those obtained by using fresh and naturally dried samples. Slightly better recoveries and precision values were obtained for the lyophilized samples. The application of the proposed methodology was tested by analyzing French bean plants from an agricultural area of Almería (Spain). The availability of this methodology, which is capable of detecting a high number of analytes in a single analysis, has priority in analyses for a large number of pesticides used on individual commodities.

Dissipation of pyrethroid residues in peppers, zucchinis, and green beans exposed to field treatments in greenhouses: evaluation by decline curves.

Dissipation of seven pyrethroid insecticides under field conditions was evaluated on green beans, zucchinis, and peppers grown in experimental greenhouses (Almería, Spain). Pyrethroid residues were determined by high performance liquid chromatography using continuous on-line post-elution photoirradiation with fluorescence detection after dichloromethane extraction and cleanup on florisil phase cartridges. Mathematically defined decline curves were established by determining optimal relationships between pyrethroid residues and time. Different models were used to find these curves. The 1st-order model achieved the best adjustment to the experimental data in 42.9% of cases. The RF (root function) 1st-order model was the best in 33.3% of times. Each of the 1.5th- and 2nd-order models provided the best adjustment in a 9.5% of the cases. Finally, the RF 1.5th-order model was the most appropriate in only 4.8% of cases. Half-life times for these three vegetables were estimated from the optimal models. The preharvest intervals for the residues in these three vegetables was obtained, taking into account the maximum residue levels established by the existing legislation. They were all lower than the ones specified by the makers of commercial formulates, which ensures a safe enough consumption.

Chiral separation of organic phosphonate compounds on cellulose CSP (chiral stationary phase) under reversed phase mode.

The enantiomers of fourteen O,O-dialkyl-2-benzyloxycarbonyl-aminoarylmethyl-phosphonates are directly separated on the tris(3,5-dimethylphenylcarbamate) cellulose column (Chiralcel OD-R) under reversed phase mode. The results of the chiral separation are different from the results obtained in the normal phase mode. The mobile phase plays an essential role in chiral discrimination when using Chiralcel OD-R. The influences of the mobile phase composition on the retention and the enantioselectivity are investigated. The influences on chiral separation of the length and steric hindrance of alkoxy groups of the phosphonate ester and of the nature of the substituent on the benzene ring that is attached to the chiral carbon atom are also discussed.

Assessment of relevant factors and relationships concerning human dermal exposure to pesticides in greenhouse applications.

Principal component analysis (PCA) was applied to the gas chromatographic data obtained from 23 different greenhouse trials. This was used to establish which factors, including application technique (very small, small, medium and large drop-size), crop characteristics (short/tall, thin/dense) and pattern application of the operator (walking towards or away from the treated area) are relevant to the dermal exposure levels of greenhouse applicators. The results showed that the highest exposure by pesticides during field applications in greenhouses, in the climatic conditions and in the crop conditions typical of a southern European country, occurs on the lower legs and front thighs of the applicators. Similar results were obtained by hierarchical cluster analysis (HCA). Drop-size seems to be very important in determining total exposure, while height and density of crops have little influence on total exposure under the conditions of the present study. No pesticide type is a major factor in total exposure. The application of multiple regression analysis (MRA) allowed assessment of the relationships between the pesticide exposure of the less affected parts of the body with the most affected parts.

Quality control criteria for analysis of organic traces in water.

The aim of this paper is the discussion of quality control (QC) criteria for environmental monitoring of organic contaminants at trace levels in water. In addition, QC criteria in the identification and confirmation of target analytes have been considered.