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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.

Hydrophilic interaction liquid chromatography is an alternative liquid chromatography mode for separation of polar compounds. In the recent years, this liquid chromatography mode has been recognized as an important solution for the analysis of compounds not amenable to reverse phase chromatography. In this work, we evaluated three different hydrophilic liquid chromatography stationary phases for the determination of 14 highly polar anionic molecules including pesticides such as glyphosate, glufosinate, ethephon and fosetyl, their main metabolites, and bromide, chlorate, and perchlorate. Several mobile phase compositions were evaluated combined with different gradients for the chromatographic run. The two columns that presented the best results were used to assess the performance for the determination of the 14 compounds in challenging highly complex feed materials. Very different matrix effects were observed for most of the compounds in each column, suggesting that different interactions can occur. Using isotopically labeled internal standards, acceptable quantitative performance and identification could be achieved down to 0.02 mg kg−1 (the lowest level tested) for most compounds. While one column was found to be favorable in terms of scope (suited for all 14 compounds), the other one was more suited for quantification and identification at lower levels, however, not for all analytes tested.

Organophosphate pesticides (OPs) are known to inhibit acetylcholine esterase (AChE), a critical effect used to establish health-based guidance values. This study developed a combined in vitro–in silico approach to predict AChE inhibition by the OP profenofos in rats and humans. A physiologically based kinetic (PBK) model was developed for both species. Parameter values for profenofos conversion to 4-bromo-2-chlorophenol (BCP) were derived from in vitro incubations with liver microsomes, liver cytosol, and plasma from rats (catalytic efficiencies of 1.1, 2.8, and 0.19 ml/min/mg protein, respectively) and humans (catalytic efficiencies of 0.17, 0.79, and 0.063 ml/min/mg protein, respectively), whereas other chemical-related parameter values were derived using in silico calculations. The rat PBK model was evaluated against literature data on urinary excretion of conjugated BCP. Concentration-dependent inhibition of rat and human AChE was determined in vitro and these data were translated with the PBK models to predicted dose-dependent AChE inhibition in rats and humans in vivo. Comparing predicted dose-dependent AChE inhibition in rats to literature data on profenofos-induced AChE inhibition revealed an accurate prediction of in vivo effect levels. Comparison of rat predictions (BMDL10 of predicted dose–response data of 0.45 mg/kg bw) and human predictions (BMDL10 of predicted dose–response data of 0.01 mg/kg bw) suggests that humans are more sensitive than rats, being mainly due to differences in kinetics. Altogether, the results demonstrate that in vivo AChE inhibition upon acute exposure to profenofos was closely predicted in rats, indicating the potential of this novel approach method in chemical hazard assessment. Supplementary Information The online version contains supplementary material available at 10.1007/s00204-021-03004-4.

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.

© 2021 The Authors. Published by Elsevier GmbH. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)., 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.

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.

A fundamental objective of the human biomonitoring for Europe initiative (HBM4EU) is to progress toward comparable and robust exposure data for a wide variety of prioritized chemicals in human samples. A programme for Quality Assurance/Quality Control (QA/QC) was designed in HBM4EU with the purpose of creating a network of European laboratories providing comparable analytical data of high quality. Two approaches were chosen for two sets of prioritized chemicals with different timelines: (i) Scheme 1, where interested candidate laboratories participated in multiple rounds of proficiency tests (ii) Scheme 2, where selected expert laboratories participated in three rounds of interlaboratory comparison investigations. In both cases, the results were used to identify laboratories capable of generating consistent and comparable results for sample analysis in the frame of HBM4EU. In total, 84 laboratories from 26 countries were invited to participate in Scheme 1 that covered up to 73 biomarkers from Hexamoll® DINCH, phthalates, bisphenols, per- and polyfluoroalkyl substances, halogenated flame retardants (HFRs), organophosporous flame retardants (OPFRs), polycyclic aromatic hydrocarbons (PAH), cadmium, chromium and aromatic amines. 74 of the participants were successful for at least one biomarker in Scheme 1. Scheme 2 involved 22 biomarkers and successful results were obtained by 2 expert laboratories for arsenic, 5 for acrylamide, 4 for mycotoxins, 2 for pesticides and 2 for UV-filters in skin care products. The QA/QC programme allowed the identification of major difficulties and needs in HBM analysis as well of gaining insight in the analytical capacities of European laboratories. Furthermore, it is the first step towards the establishment of a sustainable European network of HBM laboratories. This study was part of the HBM4EU project receiving funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No 733032. The authors acknowledge all the participating and expert laboratories (Centre de Toxicologie du Québec/Institut National de Santé Publique du Québec; IDEA Consultants, Inc.; Otsuka Pharmaceutical Co., Ltd.; Otsuka Pharmaceutical Co., Ltd.; Arizona Department of Health Services Laboratory; Centers for Disease Control and Prevention (CDC); New York State Department of Health Wadsworth Center; Wisconsin State Laboratory of Hygiene), that made the HBM4EU QA/QC programme possible, as well as the Management and Advisory Boards of HBM4EU. Sí

The world market for food supplements is large and is driven by the claims of these products to, for example, treat obesity, increase focus and alertness, decrease appetite, decrease the need for sleep or reduce impulsivity. The use of illegal compounds in food supplements is a continuous threat, certainly because these compounds and products have not been tested for safety by competent authorities. It is therefore of the utmost importance for the competent authorities to know when new products are being marketed and to warn users against potential health risks. In this study we present an approach to detect new and unknown stimulants in food supplements using machine learning. More than 20 new stimulants were identified from two different data sources, namely scientific literature applying word embedding on > 2 million abstracts and articles from formal and social media on the world wide web using text mining. The results show that the developed approach may be suitable to detect “unknowns” in the emerging risk identification activities performed by the competent authorities, which is currently a major hurdle.

Considering that pesticides have been used in Europe for over 70 years, a system for monitoring pesticide residues in EU soils and their effects on soil health is long overdue. In an attempt to address this problem, we tested 340 EU agricultural topsoil samples for multiple pesticide residues. These samples originated from 4 representative EU case study sites (CSS), which covered 3 countries and four of the main EU crops: vegetable and orange production in Spain (S-V and S-O, respectively), grape production in Portugal (P-G), and potato production in the Netherlands (N-P). Soil samples were collected between 2015 and 2018 after harvest or before the start of the growing season, depending on the CSS. Conventional and organic farming results were compared in S-V, S-O and N-P. Soils from conventional farms presented mostly mixtures of pesticide residues, with a maximum of 16 residues/sample. Soils from organic farms had significantly fewer residues, with a maximum of 5 residues/sample. The residues with the highest frequency of detection and the highest content in soil were herbicides: glyphosate and its main metabolite AMPA (P-G, N-P, S-O), and pendimethalin (S-V). Total residue content in soil reached values of 0.8 mg kg-1 for S-V, 2 mg kg-1 for S-O and N-P, and 12 mg kg-1 for P-G. Organic soils presented 70-90% lower residue concentrations than the corresponding conventional soils. There is a severe knowledge gap concerning the effects of the accumulated and complex mixtures of pesticide residues found in soil on soil biota and soil health. Safety benchmarks should be defined and introduced into (soil) legislation as soon as possible. Soil remediation techniques should be developed to keep the levels of pesticide residues below such benchmarks. Furthermore, the process of transitioning to organic farming should take into consideration the residue mixtures and their residence time in soil.

Current farm systems rely on the use of Plant Protection Products (PPP) to secure high productivity and control threats to the quality of the crops. However, PPP use may have considerable impacts on human health and the environment. A study protocol is presented aiming to determine the occurrence and levels of PPP residues in plants (crops), animals (livestock), humans and other non-target species (ecosystem representatives) for exposure modelling and impact assessment. To achieve this, we designed a cross-sectional study to compare conventional and organic farm systems across Europe. Environmental and biological samples were/are being/will be collected during the 2021 growing season, at 10 case study sites in Europe covering a range of climate zones and crops. An additional study site in Argentina will inform the impact of PPP use on growing soybean which is an important European protein-source in animal feed. We will study the impact of PPP mixtures using an integrated risk assessment methodology. The fate of PPP in environmental media (soil, water and air) and in the homes of farmers will be monitored. This will be complemented by biomonitoring to estimate PPP uptake by humans and farm animals (cow, goat, sheep and chicken), and by collection of samples from non-target species (earthworms, fish, aquatic and terrestrial macroinvertebrates, bats, and farm cats). We will use data on PPP residues in environmental and biological matrices to estimate exposures by modelling. These exposure estimates together with health and toxicity data will be used to predict the impact of PPP use on environment, plant, animal and human health. The outcome of this study will then be integrated with socio-economic information leading to an overall assessment used to identify transition pathways towards more sustainable plant protection and inform decision makers, practitioners and other stakeholders regarding farming practices and land use policy. EEA Balcarce Fil: Silva, Vera. Wageningen University and Research. Soil Physics and Land Management Group; Países Bajos. Fil: Alaoui, Abdallah. University of Bern. Institute of Geography. Centre for Development and Environment; Suiza. Fil: Schlünssen, Vivi. Aarhus University. Department of Public Health; Dinamarca. National Research Centre for the Working Environment; Dinamarca. Fil: Vested, Anne. Aarhus University. Department of Public Health; Dinamarca. Fil: Graumans, Martien. Radboud Institute for Health Sciences; Países Bajos. Fil: Van Dael, Maurice. Radboud Institute for Health Sciences; Países Bajos. Fil: Trevisan, Marco. Università Cattolica del Sacro Cuore. Department for Sustainable Food Process; Italia. Fil: Suciu, Nicoleta. Università Cattolica del Sacro Cuore. Department for Sustainable Food Process; Italia. Fil: Mol, Hans. Wageningen University and Research. Wageningen Food Safety Research; Países Bajos. Fil: Beekmann, Karsten. Wageningen University and Research. Wageningen Food Safety Research; Países Bajos. Fil: Figueiredo, Daniel. Utrecht University. Institute for Risk Assessment Sciences; Países Bajos. Fil: Harkes, Paula. Wageningen University and Research. Soil Physics and Land Management Group; Países Bajos. Fil: Hofman, Jakub. Masaryk University. Faculty of Science. Research Centre for Toxic Compounds in the Environment; Republica Checa. Fil: Kandeler, Ellen. University of Hohenheim. Institute of Soil Science and Land Evaluation; Alemania. Fil: Abrantes, Nelson. University of Aveiro. Centre for Environmental and Marine Studies and Department of Environment and Planning; Portugal. Fil: Campos, Isabel. University of Aveiro. Centre for Environmental and Marine Studies and Department of Environment and Planning; Portugal. Fil: Martínez, María Ángeles. Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas; España. Fil: Pereira, Joana Luísa. Centre for Environmental and Marine Studies and Department of Environment and Planning; Portugal. Fil: Goossens, Dirk. Wageningen University and Research. Soil Physics and Land Management Group. Países Bajos. KU Leuven Department of Earth and Environmental Sciences; Belgica. Fil: Gandrass, Juergen. Institute of Coastal Environmental Chemistry; Alemania. Fil: Debler, Freya. Institute of Coastal Environmental Chemistry; Alemania. Fil: Huerta Lwanga, Esperanza. Wageningen University and Research. Soil Physics and Land Management Group; Países Bajos. Fil: Jonker, Marlot. Dutch Mammal Society; Países Bajos. Fil: Van Langevelde, Frank. Wageningen University and Research. Wildlife Ecology and Conservation Group; Países Bajos. Fil: Sorensen, Martin. Aarhus University. Department of Animal Science; Dinamarca. Fil: Wells, Jerry. Wageningen University and Research. Host-Microbe Interactomics, Animal Sciences Group; Países Bajos. Fil: Boekhorst, Jos. Wageningen University and Research. Host-Microbe Interactomics, Animal Sciences Group; Países Bajos. Fil: Huss, Anke. Utrecht University. Institute for Risk Assessment Sciences; Países Bajos. Fil: Mandrioli, Daniele. Ramazzini Institute. Cesare Maltoni Cancer Research Center; Italia. Fil: Sgargi, Daria. Ramazzini Institute. Cesare Maltoni Cancer Research Center; Italia. Fil: Nathanail, Paul. Land Quality Management; Reino Unido. Fil: Nathanail, Judith. Land Quality Management; Reino Unido. Fil: Tamm, Lucius. Research Institute of Organic Agriculture; Suiza. Fil: Fantke, Peter. Technical University of Denmark. Department of Technology, Management and Economics; Dinamarca. Fil: Mark, Jennifer. Research Institute of Organic Agriculture; Suiza. Fil: Grovermann, Christian. Research Institute of Organic Agriculture; Suiza. Fil: Larsen, Ana Frelih. Ecologic Institute; Alemania. Fil: Herb, Irina. Ecologic Institute; Alemania. Fil: Chivers, Charlotte Anne. University of Gloucestershire. Countryside and Community Research Institute; Reino Unido. Fil: Mills, Jane. University of Gloucestershire. Countryside and Community Research Institute; Reino Unido. Fil: Alcon, Francisco. Universidad Politécnica de Cartagena; España. Fil: Contreras, Josefina. Universidad Politécnica de Cartagena; España. Fil: Baldi, Isabelle. Bordeaux University. INSERM U1219; Francia. Fil: Pasković, Igor. Institute of Agriculture and Tourism, Department of Agriculture and Nutrition; Croacia. Fil: Matjaz, Glavan. University of Ljubljana. Biotechnical Faculty; Eslovenia. Fil: Norgaard, Trine. Aarhus University. Department of Agroecology; Dinamarca. Fil: Aparicio, Virginia Carolina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Fil: Ritsema, Coen. Wageningen University and Research. Soil Physics and Land Management Group; Países Bajos. Fil: Geissen, Violette. University and Research. Soil Physics and Land Management Group; Países Bajos. Fil: Scheepers, Paul. Radboud Institute for Health Sciences; Países Bajos.

The European Human Biomonitoring Initiative (HBM4EU) is coordinating and advancing human biomonitoring (HBM). For this purpose, a network of laboratories delivering reliable analytical data on human exposure is fundamental. The analytical comparability and accuracy of laboratories analysingflame retardants(FRs) in serum and urine were investigated by a quality assurance/quality control (QA/QC) scheme comprisinginterlaboratory comparisoninvestigations (ICIs) and external quality assurance schemes (EQUASs). This paper presents the evaluation process and discusses the results of four ICI/EQUAS rounds performed from 2018 to 2020 for the determination of ten halogenated flame retardants (HFRs) represented by three congeners ofpolybrominated diphenyl ethers(BDE-47, BDE-153 and BDE-209), twoisomersof hexabromocyclododecane (α-HBCD and γ-HBCD), two dechloranes (anti-DP and syn-DP), tetrabromobisphenol A (TBBPA), decabromodiphenylethane (DBDPE), and 2,4,6-tribromophenol (2,4,6-TBP) in serum, and four metabolites of organophosphorus flame retardants (OPFRs) in urine, at two concentration levels. The number of satisfactory results reported by laboratories increased during the four rounds. In the case of HFRs, the scope of the participating laboratories varied substantially (from two to ten) and in most cases did not cover the entire target spectrum of chemicals. The highest participation rate was reached for BDE-47 and BDE-153. The majority of participants achieved more than 70% satisfactory results for these two compounds over all rounds. For other HFRs, the percentage of successful laboratories varied from 44 to 100%. The evaluation of TBBPA, DBDPE, and 2,4,6-TBP was not possible because the number of participating laboratories was too small. Only seven laboratories participated in the ICI/EQUAS scheme for OPFR metabolites and five of them were successful for at least two biomarkers. Nevertheless, the evaluation of laboratory performance using Z-scores in the first three rounds required an alternative approach compared to HFRs because of the small number of participants and the high variability of experts' results. The obtained results within the ICI/EQUAS programme showed a significant core network of comparable European laboratories for HBM of BDE-47, BDE-153, BDE-209, α-HBCD, γ-HBCD, anti-DP, and syn-DP. On the other hand, the data revealed a critically low analytical capacity in Europe for HBM of TBBPA, DBDPE, and 2,4,6-TBP as well as for the OPFR biomarkers.

Considering that pesticides have been used in Europe for over 70 years, a system for monitoring pesticide residues in EU soils and their effects on soil health is long overdue. In an attempt to address this problem, we tested 340 EU agricultural topsoil samples for multiple pesticide residues. These samples originated from 4 representative EU case study sites (CSS), which covered 3 countries and four of the main EU crops: vegetable and orange production in Spain (S–V and S–O, respectively), grape production in Portugal (P-G), and potato production in the Netherlands (N–P). Soil samples were collected between 2015 and 2018 after harvest or before the start of the growing season, depending on the CSS. Conventional and organic farming results were compared in S–V, S–O and N–P. Soils from conventional farms presented mostly mixtures of pesticide residues, with a maximum of 16 residues/sample. Soils from organic farms had significantly fewer residues, with a maximum of 5 residues/sample. The residues with the highest frequency of detection and the highest content in soil were herbicides: glyphosate and its main metabolite AMPA (P-G, N–P, S–O), and pendimethalin (S–V). Total residue content in soil reached values of 0.8 mg kg−1 for S–V, 2 mg kg−1 for S–O and N–P, and 12 mg kg−1 for P-G. Organic soils presented 70–90% lower residue concentrations than the corresponding conventional soils. There is a severe knowledge gap concerning the effects of the accumulated and complex mixtures of pesticide residues found in soil on soil biota and soil health. Safety benchmarks should be defined and introduced into (soil) legislation as soon as possible. Furthermore, the process of transitioning to organic farming should take into consideration the residue mixtures at the conversion time and their residence time in soil.

Human biomonitoring (HBM) of cadmium is essential to assess and prevent toxic exposure. Generally, low cadmium levels in urine and blood of the general population place particularly high demands on quality assurance and control measures (QA/QC) for cadmium determination. One of the aims of the HBM4EU project is to harmonize and advance HBM in Europe. Cadmium is one of the chemicals selected as a priority substance for HBM implementation in the 30 European countries under HBM4EU. For this purpose, analytical comparability and accuracy of the analytical laboratories of participating countries was investigated in a QA/QC programme comprising interlaboratory comparison investigations (ICI) and external quality assurance schemes (EQUAS). This paper presents the evaluation process and discusses the results of four ICI/EQUAS rounds for the determination of cadmium in urine and blood. The majority of the 43 participating laboratories achieved satisfactory results, although low limits of quantification were required to quantify Cd concentrations at general population exposure levels. The relative standard deviation of the participants' results obtained from all ICI and EQUAS runs ranged from 8 to 36% for cadmium in urine and 8-28% for cadmium in blood. Applying inductively-coupled plasma mass spectrometry (ICP-MS), using an internal standard, and eliminating molybdenum oxide interferences was favourable for the accurate determination of cadmium in urine and blood. Furthermore, the analysis of cadmium in urine was found to have a critical point at approximately 0.05 μg/l, below which variability increased and laboratory proficiency decreased. This QA/QC programme succeeded in establishing a network of laboratories with high analytical comparability and accuracy for the analysis of cadmium across 20 European countries. We gratefully acknowledge funding by the European Uninon's Horizon 2020 (Grant no. 733032). The authors would like to thank the HBM4EU Secretariat at the German Environment Agency for administrative support. The authors would like to express their special thanks to IDEA Consultants, Inc. (Japan), Division of Environmental Health Sciences at Wadsworth Center, New York State Department of Health, (USA), Occupational and Environmental Medicine at Laboratory Medicine, Lund (Sweden), Department of Environmental Sciences at Jožef Stefan Institute (Slovenia), Metals Laboratory – Biomarkers Unit at National Center for Environmental Health, Institute of Health Carlos III (Spain) and the laboratory team at IPASUM (Germany) for their significant support as expert laboratories. The authors acknowledge all the participating laboratories that made the HBM4EU QA/QC programme possible as well as the Management and Advisory Boards of HBM4EU. Sí

Various generic extraction methods have been used to determine pesticide residues, mycotoxins, and polycyclic aromatic hydrocarbons (PAHs) in food and animal feed to ensure consumer safety. However, these methods cannot extract all relevant compounds at an acceptable rate of recovery. This study presents a new extraction method. This new method facilitated the identification of 231 compounds, including 196 pesticides, 11 mycotoxins, and 24 PAHs over a broad range of polarities. These compounds were identified in various sample matrices, including those that are lipid-rich. The processed sample is first extracted with water, acetonitrile, formic acid, and heptane. The addition of ammonium formate results in separation into three phases and enables analysis of the aqueous phase. Solid-phase extraction clean-up procedures were performed as necessary followed by analysis by liquid or gas chromatography and mass spectrometry. Analyte recoveries were typically in the range of 70 - 120% with relative standard deviations below 20%.

The world market for food supplements is large and is driven by the claims of these products to, for example, treat obesity, increase focus and alertness, decrease appetite, decrease the need for sleep or reduce impulsivity. The use of illegal compounds in food supplements is a continuous threat, certainly because these compounds and products have not been tested for safety by competent authorities. It is therefore of the utmost importance for the competent authorities to know when new products are being marketed and to warn users against potential health risks. In this study, an approach is presented to detect new and unknown stimulants in food supplements using machine learning. More than 20 new stimulants were identified from two different data sources, namely scientific literature applying word embedding on > 2 million abstracts and articles from formal and social media on the world wide web using text mining. The results show that the developed approach may be suitable to detect “unknowns” in the emerging risk identification activities performed by the competent authorities, which is currently a major hurdle.

The current study was conducted to assess the levels of organophosphates and carbamates in vegetables in Kenya and to examine potential consumer health risks. A total of 90 samples were analysed by liquid chromatography/high-resolution tandem mass spectrometry. Residues of acephate, chlorpyrifos, methamidophos, omethoate and profenofos were found in 22% of the samples, ranging from 10 to 1343 μg/kg. The EU MRL was exceeded in 21%, 10%, 8% and 22% of the samples of French beans, kales, spinach and tomatoes, respectively. Chlorpyrifos in spinach had an acute HQ of 3.3 and 2.2 for children and adults, respectively, implying that potential health risks with respect to acute dietary exposure cannot be excluded. For chronic dietary exposure, all chronic HQs were below 1. The HI for the pesticides was 0.54 and 0.34 for children and adults. Routine monitoring of OPs and carbamates in vegetables is recommended to minimise consumer’s health risks.

Introduction: Vineyard is a crop where a large number of pesticides are applied; exposure to pesticides may occur in farmers and the general population living close to the treated area. This work aimed to investigate hair as a matrix for the assessment of cumulative and aggregate exposure to pesticides in potentially exposed individuals. Methods: Twenty agricultural workers (AW), 4 agricultural worker relatives (AR), and 5 research staff members (RS) were involved in the study. Hair samples were collected before and after the application season (PRE- and POST-EXP samples) to obtain 18 paired samples. Records with the name and the quantity of applied pesticides were obtained; twenty-seven pesticides were measured in hair by solvent extraction and LC-MS/MS. Results: During the study season, AW applied 14 different pesticides with median amount ranging from 12 to 7200 g. The most popular pesticides were dimethomorph, penconazole, cyazofamid, fenamidone and quinoxyfen, applied from 94 to 69% of AW. In AW, in PRE-EXP samples the majority of used pesticides was detectable (with detection rates from 6 to 88%), with median concentrations of few pg/mg hair; in the POST-EXP samples the frequency of detected values increased (from 25 to 100%), with median concentrations up to two orders of magnitude higher. In AR, most pesticides were quantifiable only in POST-EXP samples and with lower concentration in comparison with AW; in RS, in both PRE- and POST-EXP samples only a few pesticides were quantifiable with very low levels. In AW, a linear correlation (r = 0.682 on log-transformed data, p < 0.01) was found between the total amounts of applied pesticides during the season and their concentration in hair. Conclusion: The study shows that the majority of assessed pesticides was incorporated into hair of AW and AR. The increased frequency of detection and level at the end of the season and the correlation between pesticide in hair and the amount of applied pesticides, reinforce the use of hair for quantitative biomonitoring of cumulative exposure to pesticides.

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. Glyphosate and AMPA dynamics in the soil surface layer of cultivation areas from the loess Pampas of Argentina show high risk of AMPA accumulation, while water erosion represents a high risk for their transport to off-target areas.

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.

Viticulture, which plays a major role in the economy of several EU countries, is highly dependent of pesticides. In the São Lourenço experimental catchment, located in an important wine-growing region in Portugal, vineyards are the dominant land cover. Previous studies in this catchment showed the presence of different pesticide residues in surface water and erosion rates in vineyards of up to 30 Mg ha-1 y-1. However, information on the presence of pesticide residues in topsoils and water-eroded sediments are lacking, not just in this catchment but in general. To address these knowledge gaps, we conducted two soil sampling campaigns (fall 2015, fall 2016) and a field runoff-erosion experiment (winter 2015). We sampled 3 vineyards in the first campaign and 9 in the second, both covering different soil types (humic cambisols, calcic cambisols and chromic luvisols). Soil samples were collected within and between the vine rows, at the top, middle and bottom of the slope, and at two soil depths (0-2 and 15 cm). The runoff-erosion experiment involved 9 plots of ~12 m2; pesticide residues were monitored in topsoil and eroded sediments at six occasions after important rainfall events (>50 mm). 47 prioritized pesticide residues were analysed in 162 soil and 108 sediment samples. The levels of residues in soil were compared with their predicted environmental concentrations (PECs), calculated according to European Food Safety Authority (EFSA) pesticide application recommendations and to local farmers records. All soil samples contained pesticide residues, dimethomorph, glyphosate and its metabolite AMPA being the most frequent detected compounds. Measured levels of the different residues in soil occasionally exceeded predictions. Pesticide levels seemed related to slope position, generally being higher at the bottom than at the top or middle of the slopes, but not to the position within or between vine rows. Soil depth played a noticeable role in pesticide levels, with 80% lower concentrations of glyphosate and tebuconazole at 15 cm than 0-2 cm depth. For dieldrin (the only long-banned pesticide detected), the concentration was higher at 15 than 0-2 cm depth. Sediments had fewer compounds than soils, but the frequencies with which they were detected were similarly high. Pesticide levels were significantly higher in sediments than soils, especially in the case of the finest sediments (

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).

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).

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. The downslope off-site transport and deposition of glyphosate and AMPA by water erosion onto the soil surface can be considerable, with the consequent pollution risk of off-target terrestrial environments.

In this work, an easy and fast procedure for the selective multiresidue determination of 14 highly polar pesticides (including glyphosate, glufosinate, ethephon and fosetyl) and metabolites in beverages is presented. After an initial sample dilution (1:1, v/v), the extract is shaken and centrifuged, further diluted and then injected directly into the LC-MS/MS system, using hydrophilic interaction liquid chromatography (HILIC) and tandem mass spectrometry. No clean-up procedure was needed. The method was validated according to the current European guidelines for pesticide residue analysis in food and feed and linearity, limits of detection and quantification, matrix effects, trueness and precision were assessed. For plant-based milk, wine and beer samples, 10, 11 and 12 analytes, respectively, out of 14 were fully validated at 10 µg kg−1, the lowest spike level tested. The matrix effect was negative in most of the cases, showing for some compounds, such as HEPA, up to 80% suppression when compared to the response from standards in solvent. The use of isotopically labelled internal standards is required for the optimal quantification, as it compensates for high and varying matrix effects and also for recovery losses during extraction.

A fast screening approach for simultaneous detection of twelve pesticides and seven mycotoxins in food and feed samples using flow injection (FI) mass spectrometry (MS) was developed and evaluated. Ten selected pesticides represented five pairs of structural isomers to challenge their MS differentiation without chromatographic separation. Samples of cereals/grains and animal feed were prepared using QuEChERS extraction, diluted and fortified with the selected contaminants to yield their representative maximum levels (MLs) or maximum residue levels (MRLs). Sensitivity of the developed approach was assessed for qualitative screening of the selected contaminants using triple quadrupole MS/MS and ion mobility (IM) HR-TOFMS. Despite the high complexity of the selected matrices and the absence of chromatography, limits of detection in fortified extracts were below established regulatory levels for 14 and 15 selected contaminants by FI-MS/MS and FI-IM-TOFMS analysis, respectively, in 36–100% samples. Experiments with ion mobility TOFMS did not result in successful separation of pesticide structural isomers. However, for three out of five pairs of pesticide structural isomers: cyproconazole and uniconazole, methiocarb and ethiofencarb, and vernolate and pebulate, unique fragment ions were found and confirmed with HR-QTOFMS for their MS differentiation. The developed approach is sensitive enough for rapid multi-residue screening of some pesticides and mycotoxins in complex samples of food and feed, providing high speed and throughput, simplicity and effective use of high-end instrumentation.

Forensic examination of fire debris evidence is a notoriously difficult analytical task due to the complexity and variability of sample composition. The use of comprehensive two-dimensional gas chromatography with mass spectrometry detection (GC × GC–MS) allows the coupling of orthogonal retention mechanisms and therefore a high peak capacity.We demonstrate recent innovations in combining chemometric techniques for data reduction and feature selection, with evaluation of the evidence for forensic questions pertaining to the detection and subsequent classification of ignitable liquid residue (ILR) in fire debris samples. Chromatograms are divided into non-overlapping spatially delimited regions; for each of these regions a Local Ion Signature (LIS) is computed by summing the intensities, per nominal mass/charge over all points contained within each region. This yields a reduced feature space representing the original data as a set of consolidated ion traces. Subsequent feature selection is performed by evaluating the individual efficacy of each feature using a univariate score-based likelihood ratio (LR) approach for discriminating between pairs of same or different type samples. The retained features are used to model each ILR class using linear discriminant analysis (LDA).Results are demonstrated for 155 arson samples containing a diversity of substrate compounds and several known ignitable liquids. ILR detection is performed at 84% accuracy with fewer than 1% false positives followed by subsequent classification. Likelihood ratio distributions are presented referring to both detection and classification tasks.

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 16 mg kg− 1 of glyphosate, subjected to three SM contents (20% WHC, 60% WHC, saturation), and incubated for 30 days 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 53.5 days, while its DT90 varied between 8.0 and 280 days, depending on the treatment. AMPA persisted longer in soil than glyphosate, with its DT50 at 30 °C ranging between 26.4 and 44.5 days, and its DT90 between 87.8 and 148 days. 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.

A survey on Alternaria toxins in the food categories dried figs (n = 14), sunflower products (n = 24) and tomato products (n = 43) was carried out in the Netherlands on samples collected in retail stores in autumn 2014. The occurrence data from this survey confirmed the previously reported data of a pilot survey in 2013, with tenuazonic acid being an ubiquitous Alternaria toxin in dried figs (100% of the samples), sunflower seeds (80% of the samples) and tomato products (60% of samples) at relatively high concentrations, up to 1728 μg/kg in dried figs and 1350 μg/kg in sunflower seeds. Despite the occurring of high concentrations of tenuazonic acid in sunflower seeds and figs, it is unlikely that the population in the Netherlands is exposed to levels of concern. Alternariol was detected in 27% of the tomato products ranging from LOQ (2 μg/kg) up to 26 μg/kg, while alternariol monomethyl ether was present in 7% of the tomato products. In the worst case situation, consumption of these products may result in exposure at levels above the threshold of toxicological concern of 2.5 ng/kg body weight/day for alternariol. None of seven conjugates of Alternaria toxins was detected above the limit of quantification (LOQ) of 2.5 μg/kg.

Besides the identification point system to assure adequate set-up of instrumentation, European Commission Decision 2002/657/EC includes performance criteria regarding relative ion abundances in mass spectrometry and chromatographic retention time. In confirmatory analysis, the relative abundance of two product ions, acquired in selected reaction monitoring mode, the ion ratio should be within certain ranges for confirmation of the identity of a substance. The acceptable tolerance of the ion ratio varies with the relative abundance of the two product ions and for retention time, CD 2002/657/EC allows a tolerance of 5%. Because of rapid technical advances in analytical instruments and new approaches applied in the field of contaminant testing in food products (multi-compound and multi-class methods) a critical assessment of these criteria is justified. In this study a large number of representative, though challenging sample extracts were prepared, including muscle, urine, milk and liver, spiked with 100 registered and banned veterinary drugs at levels ranging from 0.5 to 100 µg/kg. These extracts were analysed using SRM mode using different chromatographic conditions and mass spectrometers from different vendors. In the initial study, robust data was collected using four different instrumental set-ups. Based on a unique and highly relevant data set, consisting of over 39 000 data points, the ion ratio and retention time criteria for applicability in confirmatory analysis were assessed. The outcomes were verified based on a collaborative trial including laboratories from all over the world. It was concluded that the ion ratio deviation is not related to the value of the ion ratio, but rather to the intensity of the lowest product ion. Therefore a fixed ion ratio deviation tolerance of 50% (relative) is proposed, which also is applicable for compounds present at sub-ppb levels or having poor ionisation efficiency. Furthermore, it was observed that retention time shifts, when using gradient elution, as is common practice nowadays, are mainly observed for early eluting compounds. Therefore a maximum retention time deviation of 0.2 min (absolute) is proposed. These findings should serve as input for discussions on the revision of currently applied criteria and the establishment of a new, globally accepted, criterion document for confirmatory analysis. Copyright © 2016 John Wiley & Sons, Ltd.

In 2013 the Dutch authorities issued a warning against a dietary supplement that was linked to 11 reported adverse reactions, including heart problems and in one case even a cardiac arrest. In the UK a 20-year-old woman, said to have overdosed on this supplement, died. Since according to the label the product was a herbal mixture, initial LC-MS/MS analysis focused on the detection of plant toxins. Yohimbe alkaloids, which are not allowed to be present in herbal preparations according to Dutch legislation, were found at relatively high levels (400-900 mg kg(-1)). However, their presence did not explain the adverse health effects reported. Based on these effects the supplement was screened for the presence of a β-agonist, using three different biosensor assays, i.e. the validated competitive radioligand β2-adrenergic receptor binding assay, a validated β-agonists ELISA and a newly developed multiplex microsphere (bead)-based β-agonist assay with imaging detection (MAGPIX(®)). The high responses obtained in these three biosensors suggested strongly the presence of a β-agonist. Inspection of the label indicated the presence of N-isopropyloctopamine. A pure standard of this compound was bought and shown to have a strong activity in the three biosensor assays. Analysis by LC-full-scan high-resolution MS confirmed the presence of this 'unknown known' β3-agonist N-isopropyloctopamine, reported to lead to heart problems at high doses. A confirmatory quantitative analysis revealed that one dose of the preparation resulted in an intake of 40-60 mg, which is within the therapeutic range of this compound. The case shows the strength of combining bioassays with chemical analytical techniques for identification of illegal pharmacologically active substances in food supplements.

In the past decade, several major food safety crises originated from problems with feed. Consequently, there is an urgent need for early detection of fraudulent adulteration and contamination in the feed chain. Strategies are presented for two specific cases, viz. adulterations of (i) soybean meal with melamine and other types of adulterants/contaminants and (ii) vegetable oils with mineral oil, transformer oil or other oils. These strategies comprise screening at the feed mill or port of entry with non-destructive spectroscopic methods (NIRS and Raman), followed by post-screening and confirmation in the laboratory with MS-based methods. The spectroscopic techniques are suitable for on-site and on-line applications. Currently they are suited to detect fraudulent adulteration at relatively high levels but not to detect low level contamination. The potential use of the strategies for non-targeted analysis is demonstrated.

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.

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

Rapid screening methods are currently recognized as a strategic tool for mycotoxin issues management. Specific guidelines for validation and verification of mycotoxin screening methods are set in the Commission Regulation (EU) No 2014/519. This regulation establishes that the “aim of the validation is to demonstrate the fitness-for-purpose of the screening method„ and focuses the entire validation procedure on determining specific cut-off values ensuring a maximum rate of false negative results of 5%. In addition, the assessment of the rate of false suspect results is addressed. With regard to rapid test-kits, ‘fitness-for-purpose’ includes not only the criteria more commonly considered when discussing laboratory-based methods (specificity, accuracy, and precision), but also more “practical„ parameters such as speed and ease of implementation in a new operational environment. The latter means demonstrating under local conditions that performance parameters, as established during the validation, can be achieved by first time users. This goal can be achieved through “method verification„. The aim of the present study was to verify the fitness-for-purpose of mycotoxin screening methods when applied by first time users. This was achieved in one laboratory facility via results of a training course with multiple technicians attending. The verification study was organized similarly to a collaborative exercise and involved two groups comprising of 10 technicians each that used the methods for the first time. Different screening methods were applied for deoxynivalenol (DON) in wheat, which was mainly Enzyme Linked Immunosorbent Assay (ELISA), lateral flow device (LFD), fluorescence polarization immunoassay (FPIA), and liquid chromatography-high resolution mass spectrometry (LC-HRMS). An additional verification was done for aflatoxin B1 (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.

Based on the EFSA proposal ‘Survey on sterigmatocystin in food’ (GP/EFSA/CONTAM/2013/02), this study provides a survey on the occurrence of this mycotoxin. A total of 1,259 samples of cereal grains (429), cereal products (713), beer (53) and nuts (64) were analysed for the presence of sterigmatocystin (STC). Samples were mainly collected at processing plants, storage facilities, wholesale and retail between August 2013 and November 2014, in nine European countries, mostly Greece, Italy, the Netherlands and the United Kingdom. The products originated from 27 European countries and 18 other countries. All samples were analysed by methods based on liquid chromatography coupled with tandem mass spectrometry. The limit of quantification (LOQ) was 0.5 µg/kg and the limit of detection (LOD) was in the range 0.05-0.15 µg/kg (0.005-0.01 µg/l for beer). Overall, STC was identified in 10% of the samples; it was not detected in either beer or nut samples. More than 50% of the contaminated samples contained levels between LOD and LOQ; in the other cases, levels were between 0.5-6 µg/kg with one exception (33 µg/kg in oats). In cereal grains, rice and oats seemed the cereals most prone to STC contamination (100% unprocessed rice, 22% oats grains); however the number of rice samples was limited (n=28) and the samples were collected almost exclusively in Italy and Greece. In cereal products, levels were lower than in cereal grains. The highest incidence was in processed rice (21%) and breakfast cereals (19%), while for the other cereal products this was between 5-7%. In the contaminated cereal products, rice and oats were often present as ingredients.

The decay characteristics and erosion-related transport of glyphosate and aminomethylphosphonic acid (AMPA) were monitored for 35 d at different slope gradients and rates of application in plots with loess soil on the Loess Plateau, China. The initial glyphosate decayed rapidly (half-life of 3.5d) in the upper 2 cm of soil following a first-order rate of decay. AMPA content in the 0-2 cm soil layer correspondingly peaked 3d after glyphosate application and then gradually decreased. The residues of glyphosate and AMPA decreased significantly with soil depth (p0.05) independently of the slope inclination and application rate. About 0.36% of the glyphosate initially applied was transported from plots after one erosive rain 2d after the application. Glyphosate and AMPA concentrations in runoff were low while the contents in the sediment were much higher than in the upper 2 cm of the soil.Although the rate of glyphosate decay is rapid in Chinese loess soil, the risks of glyphosate and AMPA need to be taken into account especially in the area with highly erosive rainfall.

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 0.1 mg/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 240 mg/kg. The levels of codeine and thebaine ranged from below LOQ to 348 mg/kg and from below LOQ to 106 mg/kg, respectively. Sixty percent of the samples exceeded the guidance reference value of 4 mg/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.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is one of the most widely used techniques for identification (and quantification) of residues and contaminants across a number of different chemical domains. Although the same analytical technique is used, the parameters and criteria for identification vary depending on where in the world the analysis is performed and for what purpose (e.g. determination of pesticides, veterinary drugs, forensic toxicology, sports doping). The rationale for these differences is not clear and in most cases the criteria are essentially based on expert opinions rather than underpinned by experimental data. In the current study, the variability of the two key identification parameters, retention time and ion ratio, was assessed and compared against requirements set out in different legal and guidance documents. The study involved the analysis of 120 pesticides, representing various chemical classes, polarities, molecular weights, and detector response factors, in 21 different fruit and vegetable matrices of varying degrees of complexity. The samples were analysed non-fortified, and fortified at 10, 50 and 200 μg kg(-1), in five laboratories using different LC-MS/MS instruments and conditions. In total, over 135,000 extracted-ion chromatograms were manually verified to provide an extensive data set for the assessment. The experimental data do not support relative tolerances for retention time, or different tolerances for ion ratios depending on relative abundance of the two product ions measured. Retention times in today's chromatographic systems are sufficiently stable to justify an absolute tolerance of ±0.1 min. Ion ratios are stable as long as sufficient response is obtained for both product ions. Ion ratio deviations are typically within ±20% (relative), and within ±45% (relative) in case the response of product ions are close to the limit of detection. Ion ratio tolerances up to 50% did not result in false positives and reduced the false negative rate for pesticides with product ions in the low S/N range to

A straightforward GC–MS method was developed to determine the occurrence of fourteen flavouring compounds in food. It was successfully validated for four generic types of food (liquids, semi-solids, dry solids and fatty solids) in terms of limit of quantification, linearity, selectivity, matrix effects, recovery (53–120%) and repeatability (3–22%). The method was applied to a survey of 61 Dutch food products. The survey was designed to cover all the food commodities for which the EU Regulation 1334/2008 set maximum permitted levels. All samples were compliant with EU legislation. However, the levels of coumarin (0.6–63 mg/kg) may result in an exposure that, in case of children, would exceed the tolerable daily intake (TDI) of 0.1 mg/kg bw/day. In addition to coumarin, estragole, methyl-eugenol, (R)-(+)-pulegone and thujone were EU-regulated substances detected in thirty-one of the products. The non-EU regulated alkenylbenzenes, trans -anethole and myristicin, were commonly present in beverages and in herbs-containing products.

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 2 mg kg− 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 straightforward analytical method was developed and validated to determine the mycotoxin moniliformin in cereal-based foods. Moniliformin is extracted with water and quantified with liquid chromatography tandem mass spectrometry, and its presence confirmed with liquid chromatography-Orbitrap-high-resolution mass spectrometry. The method was validated for flour, bread, pasta and maize samples in terms of linearity, matrix effect, recovery, repeatability and limit of quantification. Quantification was conducted by matrix-matched calibration. Positive samples were confirmed by standard addition. Recovery ranged from 77 to 114% and repeatability from 1 to 14%. The limit of quantification, defined as the lowest concentration tested at which the validation criteria of recovery and repeatability were fulfilled, was 10 μg/kg. The method was applied to 102 cereal-based food samples collected in the Netherlands and Germany. Moniliformin was not detected in bread samples. One of 22 flour samples contained moniliformin at 10.6 μg/kg. Moniliformin occurred in seven out of 25 pasta samples at levels around 10 μg/kg. Moniliformin (MON) was present in eight out of 23 maize products at levels ranging from 12 to 207 μg/kg. Electronic supplementary material The online version of this article (doi:10.1007/s12550-017-0287-9) contains supplementary material, which is available to authorized users.

Phomopsins (PHO) are mycotoxins produced by the fungus Diaporthe toxica (also referred to as Phomopsis leptostromiformis). Lupin is the most important host crop for this fungus and PHO are suspected as cause of lupinosis, a deadly liver disease, in sheep. Lupin is currently in use to replace genetically modified soy in many food products available on the European market. However, a validated method for analysis of PHO is not available until now. In this work, a dilute-and-shoot LC-MS/MS-based method was developed for the quantitative determination and identification of phomopsin A (PHO-A) in lupin and lupin-containing food. The method involved extraction by a mixture of acetonitrile/water/acetic acid (80/20/1 v/v), dilution of the sample in water, and direct injection of the crude extract after centrifugation. The method was validated at 5 and 25 µg PHO-A kg(-1) product. The average recovery and RSD obtained were 79% and 9%, respectively. The LOQ (the lowest level for which adequate recovery and RSD were demonstrated) was 5 µg PHO-A kg(-1). Identification of PHO-A was based on retention time and two transitions (789 > 226 and 789 > 323). Using the average of solvent standards from the sequence as a reference, retention times were all within ± 0.03 min and ion ratios were within ± 12%, which is compliant with European Union requirements. The LOD (S/N = 3 for the least sensitive transition) was 1 µg PHO-A kg(-1) product. Forty-two samples of lupin and lupin-containing food products were collected in 2011-2012 from grocery stores and internet shops in the Netherlands and analysed. In none of the samples was PHO-A detected.

A novel probabilistic Bayesian strategy is proposed to resolve highly coeluting peaks in high-resolution GC-MS (Orbitrap) data. Opposed to a deterministic approach, we propose to solve the problem probabilistically,using a complete pipeline. First, the retention time(s) for a (probabilistic) number of compounds for each mass channel are estimated. The statistical dependency between m/z channels was implied by including penalties in the model objective function. Second, Bayesian Information Criterion (BIC) is used as Occam's razor for the probabilistic assessment of the number of components. Third, a probabilistic set of resolved spectra, and their associated retention times are estimated. Finally, a probabilistic library search is proposed, computing the spectral match with a high resolution library. More specifically, a correlative measure was used that included the uncertainties in the least square fitting, as well as the probability for different proposals for the number of compounds in the mixture. The method was tested on simulated high resolution data, as well as on a set of pesticides injected in a GC-Orbitrap with high coelution. The proposed pipeline was able to detect accurately the retention times and the spectra of the peaks. For our case, with extremely high coelution situation, 5 out of the 7 existing compounds under the selected region of interest, were correctly assessed. Finally, the comparison with the classical methods of deconvolution (i.e., MCR and AMDIS) indicates a better performance of the proposed algorithm in terms of the number of correctly resolved compounds. (C) 2017 Elsevier B.V. All rights reserved.

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 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.

Plant toxins, secondary metabolites that are not essential for the survival of the organism itself but are toxic to human health, are produced by many plants. Plant toxins can be present as inherent metabolites in daily foods such as potatoes, herbs and spices or in herbal preparations. Plant toxins can be present as contaminants in foods as a result of unintentionally co-harvested weeds, transfer from feed to products of animal origin, or as residue after application of plant toxins as natural pesticides. Incidents with plant toxins are reported in Europe, mainly as a result of mixing-up of plant species used in herbal remedies or similarities between edible crops and certain weeds. EU legislation on plant toxins in food, if existing at all, is scattered over different legal acts, such as acts on contaminants, flavourings and residues. Limits for plant toxins are, furthermore, mentioned in product specifications of approved novel foods and thus related to one specific plant product. It was concluded that there is a need for a more coherent legislation on plant toxins in food in the EU to ensure consumer health.

Gas chromatography with electron ionization and full scan high resolution mass spectrometry with an Orbitrap mass analyzer (GC-EI-full scan Orbitrap HRMS) was evaluated for residue analysis. Pesticides in fruit and vegetables were taken as an example application. The relevant aspects for GC-MS based residue analysis, including the resolving power (15,000 to 120,000 FWHM at m/z 200), scan rate, dynamic range, selectivity, sensitivity, analyte identification, and utility of existing EI-libraries, are assessed and discussed in detail. The optimum acquisition conditions in full scan mode (m/z 50-500) were a resolving power of 60,000 and an automatic-gain-control target value of 3E6. These conditions provided (i) an optimum mass accuracy: within 2 ppm over a wide concentration range, with/without matrix, enabling the use of ±5 ppm mass extraction windows (ii) adequate scan speed: minimum 12 scans/peak, (iii) an intra-scan dynamic range sufficient to achieve LOD/LOQs ≤0.5 pg in fruit/vegetable matrices (corresponding to ≤0.5 μg kg(-1)) for most pesticides. EI-Orbitrap spectra were consistent over a very wide concentration range (5 orders) with good match values against NIST (EI-quadrupole) spectra. The applicability for quantitative residue analysis was verified by validation of 54 pesticides in three matrices (tomato, leek, orange) at 10 and 50 μg/kg. The method involved a QuEChERS-based extraction with a solvent switch into iso-octane, and 1 μL hot splitless injection into the GC-HRMS system. A recovery between 70 and 120% and a repeatability RSD