External Quality Assurance Schemes (EQUASs) and Inter-laboratory Comparison Investigations (ICIs) for human biomonitoring of polycyclic aromatic hydrocarbon (PAH) biomarkers in urine as part of the quality assurance programme under HBM4EU.

Polycyclic aromatic hydrocarbons (PAHs) were included as priority substances for human biomonitoring (HBM) in the European Human Biomonitoring Initiative (HBM4EU), which intended to harmonise and advance HBM across Europe. For this project, a specific Quality Assurance and Quality Control (QA/QC) programme applying Inter-laboratory Comparison Investigations (ICIs) and External Quality Assurance Schemes (EQUASs) was developed to ensure the comparability and accuracy of participating analytical laboratories. This paper presents the results of four ICI/EQUAS rounds for the determination of 13 PAH metabolites in urine, i.e. 1-naphthol, 2-naphthol, 1,2-dihydroxynaphthalene, 2-, 3- and 9-hydroxyfluorene, 1-, 2-, 3-, 4- and 9-hydroxyphenanthrene, 1-hydroxypyrene and 3-hydroxybenzo(a)pyrene. However, 4 PAH metabolites could not be evaluated as the analytical capacity of participating laboratories was too low. Across all rounds and biomarkers, 86% of the participants achieved satisfactory results, although low limits of quantification were required to quantify the urinary metabolites at exposure levels of the general population. Using high-performance liquid or gas chromatography coupled with mass spectrometry (HPLC-MS; GC-MS) and isotope dilution for calibration as well as performing an enzymatic deconjugation step proved to be favourable for the accurate determination of PAHs in urine. Finally, the HBM4EU QA/QC programme identified an international network of laboratories providing comparable results in the analysis of urinary PAH biomarkers, although covering all parameters initially selected was still too challenging.

Interlaboratory Comparison Investigations (ICIs) and External Quality Assurance Schemes (EQUASs) for human biomonitoring of perfluoroalkyl substances (PFASs) in serum as part of the quality assurance programme under HBM4EU.

Perfluoroalkyl substances (PFASs) are of very high concern due to their persistence and accumulative behaviour as well as their manifold adverse health effects. Human biomonitoring (HBM) based on the determination of PFASs in serum samples is an adequate and established strategy for exposure and risk assessment of the population. The suspected health risks associated with exposure levels in the general population call for reliable HBM data verified by Quality Assurance and Quality Control (QA/QC) measures. PFASs were among the chemicals selected as priority substances in HBM4EU, a pan-European project to harmonize and advance HBM within 30 European countries. For this purpose, the analytical comparability and accuracy of PFASs-analysing laboratories was assessed in a QA/QC programme comprising Interlaboratory Comparison Investigations (ICIs) and External Quality Assurance Schemes (EQUASs). This paper presents the evaluation process and discusses the results of four ICI/EQUAS rounds for the determination of eight perfluoroalkyl carboxylic acids and four perfluoroalkyl sulfonic acids (PFBS, PFHxS, PFHpS, PFOS) in serum. All 21 participating laboratories achieved satisfactory results for at least six of these biomarkers, although low limits of quantification (of about 0.1 µg/L) were required to quantify serum PFAS levels at general population exposure levels. The mean relative standard deviation of the participants' results (study RSDR) significantly improved from 22 % to 13 % over all PFAS biomarkers in the course of the four rounds. This QA/QC programme succeeded in establishing a network of laboratories with high analytical comparability and accuracy for the analysis of PFASs across 12 European countries.

Harmonized Quality Assurance/Quality Control Provisions for Nontargeted Measurement of Urinary Pesticide Biomarkers in the HBM4EU Multisite SPECIMEn Study.

A set of quality assurance/quality control (QA/QC) criteria for nontargeted measurement of pesticide exposure markers in a large-scale study of human urine has been proposed and applied across five laboratories within the HBM4EU project. Quality control material, including reference standards and fortified pooled urine samples (QC urine) were prepared in a centralized way and distributed across participants to monitor analytical performance and consistency of the liquid chromatography coupled to high-resolution mass spectrometry data generated with a harmonized workflow. Signal intensities, mass accuracy, and retention times of selected QA/QC markers covering a broad range of physicochemical properties were monitored across QC solvent standards, QC urine samples, study urine samples, and procedural blanks, setting acceptance thresholds for repeatability and accuracy. Overall, results showed high repeatability of the collected data. The RSDs of the signal intensities were typically below 20-30% in QC and study samples, with good stability of the chromatographic separation (retention time drift within 2-4 s intrabatch and 5 s interbatch) and excellent mass accuracy (average error < 2 ppm). The use of the proposed criteria allowed for the identification of handling errors, instrumental issues, and potential batch effects. This is the first elaboration of harmonized QA/QC criteria applied across multiple laboratories to assess the quality of data generated by nontargeted analysis of human samples.

Proficiency and Interlaboratory Variability in the Determination of Phthalate and DINCH Biomarkers in Human Urine: Results from the HBM4EU Project.

A quality assurance/quality control program was implemented in the framework of the EU project HBM4EU to assess and improve the comparability of biomarker analysis and to build a network of competent laboratories. Four rounds of proficiency tests were organized for 15 phthalate and two DINCH urinary biomarkers (0.2-138 ng/mL) over a period of 18 months, with the involvement of 28 laboratories. A substantial improvement in performance was observed after the first round in particular, and by the end of the program, an average satisfactory performance rate of 90% was achieved. The interlaboratory reproducibility as derived from the participants' results varied for the various biomarkers and rounds, with an average of 24% for the biomarkers of eight single-isomer phthalates (e.g., DnBP and DEHP) and 43% for the more challenging biomarkers of the mixed-isomer phthalates (DiNP, DiDP) and DINCH. When the reproducibility was based only on the laboratories that consistently achieved a satisfactory performance, this improved to 17% and 26%, respectively, clearly demonstrating the success of the QA/QC efforts. The program thus aided in building capacity and the establishment of a network of competent laboratories able to generate comparable and accurate HBM data for phthalate and DINCH biomarkers in 14 EU countries. In addition, global comparability was ensured by including external expert laboratories.

Quantitative assessment of multiple pesticides in silicone wristbands of children/guardian pairs living in agricultural areas in South Africa.

Little is known about personal and time-integrated exposure to past and current used pesticides in agricultural areas and within-family exposure similarities. We aimed to assess exposure to pesticides using silicone wristbands in child/guardian pairs living on farms and in villages within two agricultural areas in South Africa. Using silicone wristbands, we quantified 21 pesticides in child/guardian pairs in 38 households over six days in 2018. Levels (in ng/g wristband) of pesticides and their transformation products (12 current-use pesticides and nine organochlorine pesticides) were measured using GC-MS/MS. We assessed the correlation between pesticide levels and between household members using Spearman correlation coefficients (rs). Multivariable generalized least squares (GLS) models, using household id as intercept, were used to determine level of agreement between household members, exposure differences between children and guardians and exposure predictors (study area, household location [farm vs. village] and household pesticide use). We detected 16 pesticides with highest detection frequencies for deltamethrin (89%), chlorpyrifos (78%), boscalid (56%), cypermethrin (55%), and p,p'-DDT (48%). Most wristbands (92%) contained two or more pesticides (median seven (range one to 12)). Children had higher concentrations than guardians for four pesticides. Correlation between the pesticide levels were in most cases moderate (rs 0.30-0.68) and stronger in children than in guardians. Five pesticides showed moderate to strong correlation between household members, with the strongest correlation for boscalid (rs 0.84). Exposure differences between the two agricultural areas were observed for chlorpyrifos, diazinon, prothiofos, cypermethrin, boscalid, p,p'-DDT and p,p'-DDE and within areas for cypermethrin. We showed that for several pesticides children had higher exposure levels than guardians. The positive correlations observed for child/guardian pairs living in the same household suggest non-occupational shared exposure pathways in these communities.

A human biomonitoring (HBM) Global Registry Framework: Further advancement of HBM research following the FAIR principles.

Data generated by the rapidly evolving human biomonitoring (HBM) programmes are providing invaluable opportunities to support and advance regulatory risk assessment and management of chemicals in occupational and environmental health domains. However, heterogeneity across studies, in terms of design, terminology, biomarker nomenclature, and data formats, limits our capacity to compare and integrate data sets retrospectively (reuse). Registration of HBM studies is common for clinical trials; however, the study designs and resulting data collections cannot be traced easily. We argue that an HBM Global Registry Framework (HBM GRF) could be the solution to several of challenges hampering the (re)use of HBM (meta)data. The aim is to develop a global, host-independent HBM registry framework based on the use of harmonised open-access protocol templates from designing, undertaking of an HBM study to the use and possible reuse of the resulting HBM (meta)data. This framework should apply FAIR (Findable, Accessible, Interoperable and Reusable) principles as a core data management strategy to enable the (re)use of HBM (meta)data to its full potential through the data value chain. Moreover, we believe that implementation of FAIR principles is a fundamental enabler for digital transformation within environmental health. The HBM GRF would encompass internationally harmonised and agreed open access templates for HBM study protocols, structured web-based functionalities to deposit, find, and access harmonised protocols of HBM studies. Registration of HBM studies using the HBM GRF is anticipated to increase FAIRness of the resulting (meta)data. It is also considered that harmonisation of existing data sets could be performed retrospectively. As a consequence, data wrangling activities to make data ready for analysis will be minimised. In addition, this framework would enable the HBM (inter)national community to trace new HBM studies already in the planning phase and their results once finalised. The HBM GRF could also serve as a platform enhancing communication between scientists, risk assessors, and risk managers/policy makers. The planned European Partnership for the Assessment of Risk from Chemicals (PARC) work along these lines, based on the experience obtained in previous joint European initiatives. Therefore, PARC could very well bring a first demonstration of first essential functionalities within the development of the HBM GRF.

A health risk for consumers: the presence of adulterated food supplements in the Netherlands.

The use of food supplements is increasing. They are marketed as beneficial for health, well-being, physical or mental condition and performance, or to prevent diseases. Producers add synthetic compounds or illicit herbal material to food supplements to claim desired effects. Claims made to support marketing without scientific evidence are, however, illegal. Intake of adulterated food supplements may lead to serious adverse effects. The aim of this paper is to report the results of analyses of (adulterated) food supplements conducted by the Netherlands Food and Consumer Product Safety Authority between October 2013 and October 2018. In total, 416 supplements were analysed of which 264 (64%) contained one or more pharmacological active substances or plant toxins, such as caffeine, synephrine, sildenafil, icariin, sibutramine, higenamine, hordenine, phenethylamine, methylsynephrine, DMAA, phenolphthalein, octopamine and ephedrine. When compared to dose levels that are considered safe, daily doses of the substances in the food supplements were sometimes much higher, causing a risk for consumers who are unaware of the presence of these pharmacologically active substances. In many cases, neither food nor medicines legislation (easily) enables enforcement actions. This means that some products containing pharmacologically active substances (i.e. synthetic medicines and their illicit analogues), stay available on the market. An undesirable situation because for many of these substances no detailed toxicity data are available.

Performance Evaluation of LC-MS Methods for Multimycotoxin Determination.

The co-occurrence of regulated mycotoxins in foods and feeds, together with modified ("masked") and emerging mycotoxins, has been increasingly reported worldwide in recent years. Therefore, sensitive, accurate, and validated methods for the simultaneous determination of these hazardous contaminants in different matrices are highly demanded to fulfil regulatory requirements and to carry out reliable surveillance programs. In these last years, LC-MS methodologies for multimycotoxin screening and/or quantification are being routinely used in control laboratories. However, to date, only one European Standard for multimycotoxin determination is based on LC-MS (EN 16877:2016). The need for standardized LC-MS methods for multimycotoxin determination has been highlighted by regulatory authorities and scientific advisory bodies, including the U.S. Food and Drug Administration and the European Commission. The European Committee for Standardization (CEN) has issued calls for tender for the development of standardized LC-MS methods for mycotoxins in food and animal feeding stuffs. As deliverables, some LC-MS based methods for multimycotoxin determination are currently under approval as European Standards. In addition, the European Commission has recently established specific criteria with which screening methods for mycotoxins, including LC-MS methods, have to comply for use for regulatory purposes. Validation procedures by single-laboratory and collaborative trials have been defined. This paper provides insights and advances on guidelines and tools for performance evaluation of LC-MS methods intended for quantitative determination and for semiquantitative screening of multimycotoxins. In particular, performance criteria set in the European Union and the United States are critically overviewed, and expectations, needs, and future challenges relevant to LC-MS methods for multimycotoxin determination are also discussed.

Pesticide residues in Nepalese vegetables and potential health risks.

We conducted this study in order to assess the pesticide residues in vegetables and examine the related human health risk. Therefore, residues of 23 pesticides (organophosphates, organochlorines, acaricides, fungicides, and insecticides of biological origin) were analysed in the three main vegetable crops grown in Southern Nepal: 27 eggplant, 27 chilli and 32 tomato samples representing (i) conventional (N = 67) and ii) integrated pest management (IPM) fields (N = 19). Pesticide residues were found in 93% of the eggplant samples and in all of the chilli and tomato samples. Multiple residues were observed in 56% of the eggplant samples, 96% of chilli samples and all of the tomato samples. The range (µg/kg) of total detected pesticide residues in eggplants, chillies and tomatoes was 1.71-231, 4.97-507, 13.1-3465, respectively. The most frequently detected pesticides in these vegetables were carbendazim and chloropyrifos. Pesticide residues in 4% of the eggplant, 44% of the tomato and 19% of the chilli samples exceeded the EU maximum residue limits (MRLs). The residues of triazophos, omethoate, chloropyrifos and carbendazim exceeded the EU MRLs. Compared to chilli and eggplant crops, more carbendazim was sprayed onto tomato crops (p < 0.05). We assessed adolescent and adult dietary exposure using hazard quotient (HQ) and hazard index (HI) equations for the identified pesticides. HQ> 1 was observed for chloropyrifos, triazophos and carbendazim in eggplants; profenofos, triazophos, dimethoate, omethoate, chloropyrifos and carbendazim in tomatoes; and dichlorvos and chloropyrifos in chillies. Of all of the HQs, the highest acute HQ (aHQ) was for triazophos (tomato) in adolescents (aHQ=657) and adults (aHQ=677), showing the highest risks of dietary exposure. The cumulative dietary exposure showed a higher HI for organophosphates (HI>83) and a lower HI for organochlorines, acaricides and biological insecticides (HI<1). The concentration of pesticide residues in the vegetable crops from the IPM field was considerably lower, suggesting a greater ability of IPM systems to reduce the dietary risks from exposure to pesticides.

Pesticide residues in European agricultural soils - A hidden reality unfolded.

Pesticide use is a major foundation of the agricultural intensification observed over the last few decades. As a result, soil contamination by pesticide residues has become an issue of increasing concern due to some pesticides' high soil persistence and toxicity to non-target species. In this study, the distribution of 76 pesticide residues was evaluated in 317 agricultural topsoil samples from across the European Union. The soils were collected in 2015 and originated from 11 EU Member States and 6 main cropping systems. Over 80% of the tested soils contained pesticide residues (25% of samples had 1 residue, 58% of samples had mixtures of two or more residues), in a total of 166 different pesticide combinations. Glyphosate and its metabolite AMPA, DDTs (DDT and its metabolites) and the broad-spectrum fungicides boscalid, epoxiconazole and tebuconazole were the compounds most frequently found in soil samples and the compounds found at the highest concentrations. These compounds occasionally exceeded their predicted environmental concentrations in soil but were below the respective toxic endpoints for standard in-soil organisms. Maximum individual pesticide content assessed in a soil sample was 2.05 mg kg-1 while maximum total pesticide content was 2.87 mg kg-1. This study reveals that the presence of mixtures of pesticide residues in soils are the rule rather than the exception, indicating that environmental risk assessment procedures should be adapted accordingly to minimize related risks to soil life and beyond. This information can be used to implement monitoring programs for pesticide residues in soil and to trigger toxicity assessments of mixtures of pesticide residues on a wider range of soil species in order to perform more comprehensive and accurate risk assessments.

Evaluation of Mycotoxin Screening Tests in a Verification Study Involving First Time Users.

(AFB1) in maize and wheat using LFD and LC-HRMS, respectively. The results of analyses were used to calculate intermediate precision (RSDip, covering the inter-analyst variability in preparing the analytical samples and the precision under repeatability conditions) cut-off values and false suspect rates. RSDip ranged from 6.5% to 30% for DON, and from 16% to 33% for AFB1. The highest obtained variances were associated with the AFB1 analyses due to working with much lower mass fractions. The rate of false suspect results were lower than 0.1% for all tested methods. All methods showed a fit-for-purpose method performance profile, which allowed a clear distinction of samples containing the analytes at the screening target concentration (STC) from negative control samples. Moreover, the first time users obtained method performances similar to those obtained for validation studies previously performed on the screening methods included in the training course.

Dynamics of glyphosate and AMPA in the soil surface layer of glyphosate-resistant crop cultivations in the loess Pampas of Argentina.

This study investigates the dynamics of glyphosate and AMPA in the soil surface layer of two fields growing glyphosate-resistant crops in the loess Pampas of Córdoba Province, Argentina. Glyphosate decay and AMPA formation/decay were studied after a single application, using decay kinetic models. Furthermore, glyphosate and AMPA concentrations were investigated in runoff to evaluate their off-site risk. During a 2.5-month study, cultivations of glyphosate-resistant soybean and maize received an application of 1.0 and 0.81 kg a.e. ha-1, respectively, of Roundup UltraMax©. Topsoil samples (0-1, 1-2 cm) were collected weekly (including before application) and analysed for glyphosate, AMPA and soil moisture (SM) contents. Runoff was collected from runoff plots (3 m2) and weirs after 2 erosive rainfall events, and analysed for glyphosate and AMPA contents (water, eroded-sediment). Under both cultivations, background residues in soil before application were 0.27-0.42 mg kg-1 for glyphosate and 1.3-1.7 mg kg-1 for AMPA. In the soybean area, the single-first-order (SFO) model performed best for glyphosate decay. In the maize area, the bi-phasic Hockey-Stick (HS) model performed best for glyphosate decay, due to an abrupt change in SM regimes after high rainfall. Glyphosate half-life and DT90 were 6.0 and 19.8 days, respectively, in the soybean area, and 11.1 and 15.4 days, respectively, in the maize area. In the soybean area, 24% of the glyphosate was degraded to AMPA. In the maize area, it was only 5%. AMPA half-life and DT90 were 54.7 and 182 days, respectively, in the soybean area, and 71.0 and 236 days, respectively, in the maize area. Glyphosate and AMPA contents were 1.1-17.5 times higher in water-eroded sediment than in soil. We conclude that AMPA persists and may accumulate in soil, whereas both glyphosate and AMPA are prone to off-site transport with water erosion, representing a contamination risk for surface waters and adjacent fields.

Spatial glyphosate and AMPA redistribution on the soil surface driven by sediment transport processes - A flume experiment.

This study investigates the influence of small-scale sediment transport on glyphosate and AMPA redistribution on the soil surface and on their off-site transport during water erosion events. Both a smooth surface (T1) and a surface with "seeding lines on the contour" (T2) were tested in a rainfall simulation experiment using soil flumes (1 × 0.5 m) with a 5% slope. A dose of 178 mg m-2 of a glyphosate-based formulation (CLINIC®) was applied on the upper 0.2 m of the flumes. Four 15-min rainfall events (RE) with 30-min interval in between and a total rainfall intensity of 30 mm h-1 were applied. Runoff samples were collected after each RE in a collector at the flume outlet. At the end of the four REs, soil and sediment samples were collected in the application area and in four 20 cm-segments downslope of the application area. Samples were collected according to the following visually distinguished soil surface groups: light sedimentation (LS), dark sedimentation (DS), background and aggregates. Results showed that runoff, suspended sediment and associated glyphosate and AMPA off-site transport were significantly lower in T2 than in T1. Glyphosate and AMPA off-site deposition was higher for T2 than for T1, and their contents on the soil surface decreased with increasing distance from the application area for all soil surface groups and in both treatments. The LS and DS groups presented the highest glyphosate and AMPA contents, but the background group contributed the most to the downslope off-site deposition. Glyphosate and AMPA off-target particle-bound transport was 9.4% (T1) and 17.8% (T2) of the applied amount, while water-dissolved transport was 2.8% (T1) and 0.5% (T2). Particle size and organic matter influenced the mobility of glyphosate and AMPA to off-target areas. These results indicate that the pollution risk of terrestrial and aquatic environments through runoff and deposition can be considerable.

Enhancing detectability of anabolic-steroid residues in bovine urine by actively modulated online comprehensive two-dimensional liquid chromatography - high-resolution mass spectrometry.

In this study we describe an approach to enhance the sensitivity of an online comprehensive two-dimensional liquid chromatography (LC × LC) high-resolution mass spectrometry method for the separation and detection of trace levels of anabolic-steroid residues in complex urine matrices. Compared to one-dimensional liquid chromatography (1D-LC), LC × LC methods offer higher separation power, thanks to the combined effect of two different selectivities and a higher peak capacity. However, when using state-of-the-art LC × LC instrumentation, the price paid for the increase in separation power is a decrease in sensitivity and detectability of trace-level analytes. This can be ascribed to the sample dilution that takes place during each of the two chromatographic steps. The way in which fractions are collected and transferred from the first to the second column is also of paramount importance, especially the volume and the solvent composition of the fractions injected in the second column. To overcome the detection limitation, we present an active-modulation strategy, based on concentrating the fractions of the first-dimension effluent using a modulation interface that employs trap columns. We obtained a signal enhancement for anabolic-steroid compounds in a bovine-urine sample by a factor of 2.4-7.6 and an increase in the signal-to-noise ratio up to a factor of 7 in comparison with a standard loop-based modulation interface. In addition, thanks to the increased sensitivity of our method, a substantially larger number of peaks were detected (76 vs. 36). Moreover, we could reduce the solvent consumption by a factor of three (160 mL vs. 500 mL per run).

Application of Gas Chromatography Coupled to Quadrupole-Orbitrap Mass Spectrometry for Pesticide Residue Analysis in Cereals and Feed Ingredients.

A method for residue analysis of pesticides and polychlorinated biphenyls in cereals and feed ingredients based on QuEChERS extraction, programmed temperature vaporizer large-volume injection, and GC with electron ionization (EI) quadrupole Orbitrap full-scan high-resolution MS (60 000 full width at half-maximum at m/z 200) has been developed. In addition to full-scan acquisition, simultaneous full-scan and selected-ion monitoring acquisition was used to improve detectability in incidental cases in which analytes coeluted with intense signals from coextractants. The method was successfully validated down to 10 µg/kg for a single commodity (wheat) using matrix-matched calibration, and for multiple-feed matrixes using standard addition. Identification according to European Union requirements was achieved in >90% of the analyte/matrix combinations, and suggestions for further increasing identification rates have been made. Performance characteristics were compared to an existing method for residue analysis based on GC with EI tandem MS (triple quadrupole).

Straightforward analytical method to determine opium alkaloids in poppy seeds and bakery products.

A straightforward method to determine the content of six opium alkaloids (morphine, codeine, thebaine, noscapine, papaverine and narceine) in poppy seeds and bakery products was developed and validated down to a limit of quantification (LOQ) of 0.1mg/kg. The method was based on extraction with acetonitrile/water/formic acid, ten-fold dilution and analysis by LC-MS/MS using a pH 10 carbonate buffer. The method was applied for the analysis of 41 samples collected in 2015 in the Netherlands and Germany. All samples contained morphine ranging from 0.2 to 240mg/kg. The levels of codeine and thebaine ranged from below LOQ to 348mg/kg and from below LOQ to 106mg/kg, respectively. Sixty percent of the samples exceeded the guidance reference value of 4mg/kg of morphine set by BfR in Germany, whereas 25% of the samples did not comply with the limits set for morphine, codeine, thebaine and noscapine by Hungarian legislation.

Distribution of glyphosate and aminomethylphosphonic acid (AMPA) in agricultural topsoils of the European Union.

Approval for glyphosate-based herbicides in the European Union (EU) is under intense debate due to concern about their effects on the environment and human health. The occurrence of glyphosate residues in European water bodies is rather well documented whereas only few, fragmented and outdated information is available for European soils. We provide the first large-scale assessment of distribution (occurrence and concentrations) of glyphosate and its main metabolite aminomethylphosphonic acid (AMPA) in EU agricultural topsoils, and estimate their potential spreading by wind and water erosion. Glyphosate and/or AMPA were present in 45% of the topsoils collected, originating from eleven countries and six crop systems, with a maximum concentration of 2mgkg-1. Several glyphosate and AMPA hotspots were identified across the EU. Soil loss rates (obtained from recently derived European maps) were used to estimate the potential export of glyphosate and AMPA by wind and water erosion. The estimated exports, result of a conceptually simple model, clearly indicate that particulate transport can contribute to human and environmental exposure to herbicide residues. Residue threshold values in soils are urgently needed to define potential risks for soil health and off site effects related to export by wind and water erosion.

A global inter-laboratory study to assess acquisition modes for multi-compound confirmatory analysis of veterinary drugs using liquid chromatography coupled to triple quadrupole, time of flight and orbitrap mass spectrometry.

According to EU legislation a confirmatory method used for residue analysis should be able to confirm the identity of a compound beyond reasonable doubt. To provide an adequate instrumental set-up, Commission Decision 2002/657/EC introduced the concept of "identification points". A second aspect to assure unequivocal confirmation, is the establishment of ion ratio and retention time criteria. Currently, the gold standard for confirmatory analysis of most veterinary drug residues is liquid chromatography (LC) coupled to tandem mass spectrometry (MS/MS) in selected reaction monitoring (SRM) acquisition mode, isolating one precursor ion and monitoring two a priori selected product ions, yielding 4 identification points. We comprehensively evaluated the use of different low and high resolution LC-MS(/MS) techniques and acquisition modes with respect to the selectivity of 100 veterinary drugs in liver, muscle and urine extracts aiming to critically review the currently established identification points system. A comparison among MS/MS in SRM mode with high resolution mass spectrometry (HRMS) in full scan, all ion fragmentation and targeted MS/MS was made based on a unique inter-laboratory study, which comprises 21 laboratories from four different continents and equipment from all major vendors. In total 186 samples were analysed yielding results for 9282 analyte/matrix combinations. It was observed that the false positive rate approximately doubles if no ion ratio criterion is applied indicating that this criterion is important to prevent false positive results. Full scan HRMS analysis, only monitoring the molecular ion and allowing a ±5 ppm mass tolerance is, in general, less selective than low resolution MS/MS using SRM, and thus full scan alone is considered not sufficient for confirmatory analysis. Furthermore, even though the number of data on all ion fragmentation and targeted MS/MS at high resolution was limited, based on the data obtained, it was observed that the acquisition mode as well as the mass resolution needed, very much depend on the matrix and the compound itself. For complex matrix extracts and non-selective compounds (worst-case situation), only targeted MS/MS, monitoring the precursor ion and a single product ion in HR-MS using a maximum of ±5 ppm mass deviation, leads to comparable selectivity and false positive and negative rate as SRM monitoring two product ions in LR-MS. We conclude that the currently applied identification point system as established in commission decision 2002/657/EC should be revised with respect to the allocation of identification points.

Glyphosate and AMPA distribution in wind-eroded sediment derived from loess soil.

Glyphosate is one of the most used herbicides in agricultural lands worldwide. Wind-eroded sediment and dust, as an environmental transport pathway of glyphosate and of its main metabolite aminomethylphosphonic acid (AMPA), can result in environmental- and human exposure far beyond the agricultural areas where it has been applied. Therefore, special attention is required to the airborne transport of glyphosate and AMPA. In this study, we investigated the behavior of glyphosate and AMPA in wind-eroded sediment by measuring their content in different size fractions (median diameters between 715 and 8 µm) of a loess soil, during a period of 28 days after glyphosate application. Granulometrical extraction was done using a wind tunnel and a Soil Fine Particle Extractor. Extractions were conducted on days 0, 3, 7, 14, 21 and 28 after glyphosate application. Results indicated that glyphosate and AMPA contents were significantly higher in the finest particle fractions (median diameters between 8 and 18 µm), and lowered significantly with the increase in particle size. However, their content remained constant when aggregates were present in the sample. Glyphosate and AMPA contents correlated positively with clay, organic matter, and silt content. The dissipation of glyphosate over time was very low, which was most probably due to the low soil moisture content of the sediment. Consequently, the formation of AMPA was also very low. The low dissipation of glyphosate in our study indicates that the risk of glyphosate transport in dry sediment to off-target areas by wind can be very high. The highest glyphosate and AMPA contents were found in the smallest soil fractions (PM10 and less), which are easily inhaled and, therefore, contribute to human exposure.

Persistence of glyphosate and aminomethylphosphonic acid in loess soil under different combinations of temperature, soil moisture and light/darkness.

The dissipation kinetics of glyphosate and its metabolite aminomethylphosphonic acid (AMPA) were studied in loess soil, under biotic and abiotic conditions, as affected by temperature, soil moisture (SM) and light/darkness. Nonsterile and sterile soil samples were spiked with 16mgkg-1 of glyphosate, subjected to three SM contents (20% WHC, 60% WHC, saturation), and incubated for 30days at 5°C and 30°C, under dark and light regimes. Glyphosate and AMPA dissipation kinetics were fit to single-first-order (SFO) or first-order-multicompartment (FOMC) models, per treatment combination. AMPA kinetic model included both the formation and decline phases. Glyphosate dissipation kinetics followed SFO at 5°C, but FOMC at 30°C. AMPA followed SFO dissipation kinetics for all treatments. Glyphosate and AMPA dissipation occurred mostly by microbial activity. Abiotic processes played a negligible role for both compounds. Under biotic conditions, glyphosate dissipation and AMPA formation/dissipation were primarily affected by temperature, but also by SM. Light regimes didn't play a significant role. Glyphosate DT50 varied between 1.5 and 53.5days, while its DT90 varied between 8.0 and 280days, depending on the treatment. AMPA persisted longer in soil than glyphosate, with its DT50 at 30°C ranging between 26.4 and 44.5days, and its DT90 between 87.8 and 148days. The shortest DT50/DT90 values for both compounds occurred at 30°C and under optimal/saturated moisture conditions, while the largest occurred at 5°C and reaching drought stress conditions. Based on these results, we conclude that glyphosate and AMPA dissipate rapidly under warm and rainy climate conditions. However, repeated glyphosate applications in fallows or winter crops in countries where cold and dry winters normally occur could lead to on-site soil pollution, with consequent potential risks to the environment and human health. To our knowledge, this study is the first evaluating the combined effect of temperature, soil moisture and light/dark conditions on AMPA formation/dissipation kinetics and behaviour.