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Monitoring the exposure to ethoxyquin between 2000 and 2021 in urine samples from the German Environmental Specimen Bank

Human Biomonitoring (HBM) of emerging chemicals gained increasing attention within the EU in recent years. After evaluating the metabolism, we established a new HBM method for ethoxyquin (EQ), a feed additive, which was banned in 2017 due to concerns regarding the possible exposure of the general population to it and its highly toxic precursor p-phenetidine. The method was applied to 250 urine samples from the Environmental Specimen Bank collected between 2000 and 2021. The major metabolite EQI was quantified in the majority of the study samples illustrating the ubiquitous exposure of the non-occupationally exposed population. A rather constant exposure was observed until 2016 with a significant decline from 2016 to 2021. This drop falls within the EU wide ban of the chemical as a feed additive from June 2017 which led to a gradual removal until its complete suspension in June 2020. The daily intake (DI) was evaluated with respect to the reported derived no-effect level (DNEL) to estimate the potential health risks from EQ exposure. The median DI of 0.0181 Ìg/kg bw/d corresponds to only 0.01 % of the DNEL. Even the observed maxima up to 13.1 Ìg/kg bw/d only accounted for 10 % of the DNEL. Nevertheless, the values suggest a general exposure with the risk of higher burden in a low fraction of the population. In regard to the EQ associated intake of the carcinogen and suspected mutagen p-phenetidine, this level of exposure cannot be evaluated as safe. The recent decrease and the broad exposure substantiate the need for future HBM campaigns in population representative studies to further investigate the observed reductions, potentially find highly exposed subgroups and clarify the impact of the ban as feed additive on EQ exposure. © 2023 The Author(s)

A time trend of urinary 4-methylbenzylidene camphor metabolites in young adults from Germany

4-methylbenzylidene camphor (4-MBC) is used as a UV-B filter in cosmetics. Two oxidized metabolites of 4-MBC - 3-(4-carboxybenzylidene)camphor (cx-MBC) and 3-(4-carboxybenzylidene)-6-hydroxycamphor (cx-MBC-OH) - were analyzed in 250 24-h urine samples from young adults in Germany. The samples were from the German Environmental Specimen Bank (ESB) and represented exposure in the years 1995, 2005, 2010, 2015 and 2019. A UHPLC-MS/MS method enabled the sensitive determination of both metabolites, with limits of quantifi cation at 0.15 mikrogL-1 (cx-MBC) and 0.30 mikrogL-1 (cx-MBC-OH), respectively. A temporal trend of the internal exposure to 4-MBC was clearly noticeable. The metabolite cx-MBC was frequently quantifiable at the beginning of the period: in 70% of the samples in 1995, and 56% in 2005. After 2005, urinary concentrations and detection rates of cx-MBC dropped to reach very low levels. In 2015 and 2019, the detection rate was only 2% and 0%, respectively. A similar trend was observed for cx-MBC-OH, though overall, this metabolite was detected less often and at lower concentration levels than cx-MBC. Nowadays, measurable levels of urinary 4-MBC metabolites are an extremely rare occurrence in Germany. These trends are consistent with the history of 4-MBC use by the cosmetic industry. The highest measured individual concentration of 16.20 mikrogL-1 (in a sample of the year 2005) was still more than 30 times below the health-based guidance value (HBM-I). An investigation of the ratios between both metabolites uncovered several features of the 4-MBC metabolism which have been essentially overlooked until now. In particular, stereochemical aspects should be explored in future studies. As urine was collected in autumn/winter in Northwestern Germany, the 4-MBC metabolites measured prob ably do not arise from sunscreen products in a narrow sense. They rather may reveal the use of other skin care products containing 4-MBC for UV protection as an added feature. © 2023 Elsevier

Human urinary arsenic species, associated exposure determinants and potential health risks assessed in the HBM4EU Aligned Studies

The European Joint Programme HBM4EU coordinated and advanced human biomonitoring (HBM) in Europe in order to provide science-based evidence for chemical policy development and improve chemical management. Arsenic (As) was selected as a priority substance under the HBM4EU initiative for which open, policy relevant questions like the status of exposure had to be answered. Internal exposure to inorganic arsenic (iAs), measured as Toxic Relevant Arsenic (TRA) (the sum of As(III), As(V), MMA, DMA) in urine samples of teenagers differed among the sampling sites (BEA (Spain) > Riksmaten adolescents (Sweden), ESTEBAN (France) > FLEHS IV (Belgium), SLO CRP (Slovenia)) with geometric means between 3.84 and 8.47 mikrog/L. The ratio TRA to TRA + arsenobetaine or the ratio TRA to total arsenic varied between 0.22 and 0.49. Main exposure determinants for TRA were the consumption of rice and seafood. When all studies were combined, Pearson correlation analysis showed significant associations between all considered As species. Higher concentrations of DMA, quantitatively a major constituent of TRA, were found with increasing arsenobetaine concentrations, a marker for organic As intake, e.g. through seafood, indicating that other sources of DMA than metabolism of inorganic As exist, e.g. direct intake of DMA or via the intake of arsenosugars or -lipids. Given the lower toxicity of DMA(V) versus iAs, estimating the amount of DMA not originating from iAs, or normalizing TRA for arsenobetaine intake could be useful for estimating iAs exposure and risk. Comparing urinary TRA concentrations with formerly derived biomonitoring equivalent (BE) for non-carcinogenic effects (6.4 mikrog/L) clearly shows that all 95th percentile exposure values in the different studies exceeded this BE. This together with the fact that cancer risk may not be excluded even at lower iAs levels, suggests a possible health concern for the general population of Europe. © 2023 The Authors

Influence of low oxygen concentrations on biological transformations of trace organic chemicals in sand filter systems

Managed aquifer recharge systems for drinking water reclamation are challenged by trace organic chemicals (TOrCs) since some of them are poorly retained. Although a lot of research has been done to investigate biological transformation of TOrCs in sand filter systems, there are still uncertainties to predict the removal. A laboratory column system with two different filter sands was set up to test TOrC transformation, the influence of low oxygen concentrations as well as the adaptation and influence of spiked TOrC influent concentrations. Bioactivity was quantified with the fluorescence tracer resazurin. In the experiment, a low elimination performance in the first column segment, defined as lag zone, was observed, implying incomplete adaptation or inhibiting co-factors. To assess these lag zones and to determine the dissipation time DT50 for 50% removal, a modified Gompertz model was applied. For acesulfame, formylaminoantipyrine, gabapentin, sulfamethoxazole, and valsartan acid DT50 of less than 10 h were observed, even when influent oxygen concentrations decreased to 0.5 mg/L. In general, TOrC transformations in technical sand with lower bioactivity and especially valsartan acid transformation responded very sensitive to low influent oxygen concentrations of 0.5 mg/L. However, in well adapted sand originating from soil aquifer treatment (SAT) with sufficient bioactivity, TOrC removal was hardly affected by such suboxic conditions. Furthermore, increasing the influent concentrations of TOrCs to 10 ÎÌg/L was found to promote adaptation especially for acesulfame and sulfamethoxazole. Benzotriazole, carbamazepine, diclofenac and venlafaxine were recalcitrant under the applied experimental conditions. © 2023 Elsevier

Glyphosate and AMPA in human urine of HBM4EU-aligned studies: Part A children

Few data are available on the exposure of children to glyphosate (Gly) in Europe. Within HBM4EU, new HBM exposure data were collected from aligned studies at five sampling sites distributed over Europe (studies: SLO CRP (SI); ORGANIKO (CY); GerES V-sub (DE); 3XG (BE); ESTEBAN (FR)). Median Gly concentrations in urine were below or around the detection limit (0.1 Ìg/L). The 95th percentiles ranged between 0.18 and 1.03 Ìg Gly/L. The ratio of AMPA (aminomethylphosphonic acid; main metabolite of Gly) to Gly at molar basis was on average 2.2 and the ratio decreased with higher Gly concentrations, suggesting that other sources of AMPA, independent of metabolism of Gly to AMPA in the monitored participants, may concurrently operate. Using reverse dosimetry and HBM exposure data from five European countries (east, west and south Europe) combined with the proposed ADI (acceptable daily intake) of EFSA for Gly of 0.1 mg/kg bw/day (based on histopathological findings in the salivary gland of rats) indicated no human health risks for Gly in the studied populations at the moment. However, the absence of a group ADI for Gly+AMPA and ongoing discussions on e.g., endocrine disrupting effects cast some uncertainty in relation to the current single substance ADI for Gly. The carcinogenic effects of Gly are still debated in the scientific community. These outcomes would influence the risk conclusions presented here. Finally, regression analyses did not find clear associations between urinary exposure biomarkers and analyzed potential exposure determinants. More information from questionnaires targeting exposure-related behavior just before the sampling is needed. © 2022 by the authors

New insights on in vitro biotransformation of anticoagulant rodenticides in fish

he assessment of the bioaccumulation potential of chemicals is an essential and mandatory part of their regulatory environmental risk and hazard assessment. So far, in vitro data on fish metabolism is rarely available for biocidal active substances such as anticoagulant rodenticides. In this case study we present in vitro biotransformation rates of eight biocidal and one pharmaceutical anticoagulants in rainbow trout (Oncorhynchus mykiss) liver subcellular S9 fraction (RT-S9) determined following the Organisation for Economic Co-operation and Development test guideline 319B method at two different incubation temperatures (i.e., 12 +/- 1 ËÌC and 23 +/- 2 ËÌC). Furthermore, we address challenges associated with the usability and interpretation of in vitro data to support the decision making within the regulatory bioaccumulation assessment in bridging the gap between in silico methods and in vivo studies. According to our results, four of the tested substances (i.e., chlorophacinone, coumatetralyl, bromadiolone, and difenacoum) exhibited significant intrinsic clearance (p < .001) in the RT-S9 assay. Overall, the observed metabolism was (very) slow and clearance rates were temperature-dependent. Whether the determined in vitro biotransformation rate had a substantial influence on the predicted bioconcentration factor during extrapolation was subject to the lipophilicity of the test substance. Further improvements of existing concepts are needed to overcome uncertainties in the prediction of bioconcentration factors for chemicals such as anticoagulants. © 2022 Elsevier Ltd.

Monitoring framework for the use of natural resources in Germany

Indicators are required to monitor the progress of resource and circular economy policies. The German Sustainable Development Strategy and Resource Efficiency Program already include a number of indicators for mapping Germany's resource use and socio-economic metabolism. However, currently used indicators only include a subset of natural resources and often lack an impact evaluation (e.g., considering resource scarcity or environmental relevance). Resource and environmental footprints indirectly caused through the trade of goods have so far only partly been considered by German resource policy and in official statistics. As a result, burden shifting between different resource categories, world regions, or environmental effects can remain undetected. To fill this gap, we discuss the overall scope of natural resource monitoring in Germany and review existing resource indicators evaluating them against a set of predefined criteria. We then propose a possible monitoring framework for Germany consisting of a materials-layer (the focus of resource and circular economy policies to date) for the evaluation of material flows and stocks, and corresponding water, land, and emissions-layers which should be monitored simultaneously to track contributions to the overarching objectives of resource and circular economy policies. Possible indicators and data sources are discussed and an outlook for future research provided. © 2021 The Authors

Sediment contamination of an urban canal - a case study approach for an integrated assessment of organic sediment contamination

Purpose The pesticide DDT and its metabolites represent a contamination risk for the aquatic environment, especially the polar metabolite DDA. The study provides a quantitative assessment of long-term pollution risks from sedimentary DDT residues with a special focus on DDA. It presents an overview of the contamination range of different DDX compounds in the sediments of a canal in Berlin (Germany), resulting from a former industrial point source that has implications for drinking water resources in the nearby area. The comprehensive analysis scheme provides information on free accessible and potentially metabolized precursors also in the non-extractable residues. This allows a quantitative assessment of the DDA pollution potential derived from the sedimentary DDT residues. Materials and methods The area was investigated for fine-grained sediment by means of a geo-electric mapping. Twelve sediment cores were taken in four areas (three in each section). A wider range of precursor metabolites has been included due to their transformation potential to the polar metabolite DDA. The sediments were analysed quantitatively for extractable and easily releasable fractions by application of a variety of degradation techniques as well as a dispersion extraction procedure on the sediment samples. These extracts were fractionated and subsequently analysed by GC-MS. Results and discussion Concentrations were obtained for extractable and bound metabolites. Different scenarios for the calculation of the amount of contaminated sediment are displayed as a tool for contamination assessment. The formation potential of DDA as the water-soluble metabolite is presented. Several precursor metabolites, e.g. DDD and DDMS, extractable from the sediment organic matter, revealed a high potential for a long-term formation of DDA, especially in the easily releasable fraction (via hydrolysis) with a mean concentration of up to 11,000 (micro)g g-1 dry sediment. The resulting DDA contamination potential represents a significant pollution risk for the groundwater from a downstream waterworks area and by remobilisation into the whole ecosystem and adjacent rivers. Conclusions The application of the presented methods provides a tool for a quantitative assessment of the long-term release potential of DDA under different scenarios by a comprehensive analysis of contaminated sediments (and soils). This approach can be transferred to pollutants that are also characterized by a complex metabolism accompanied by bound residue formation. © The Author(s) 2020

A biomonitoring study assessing the exposure of young German adults to butylated hydroxytoluene (BHT)

The antioxidant 2,6-di-tert-butyl-4-methylphenol (butylated hydroxytoluene, BHT) is used ubiquitously in food, cosmetics, pharmaceuticals, fuels, plastics, rubbers and many other products. Therefore, exposure of the general population to this substance is likely. We analyzed the BHT metabolite 3,5-di-tert-butyl-4-hydroxybenzoic acid ("BHT acid") in 24-h urine samples from the German Environmental Specimen Bank with the aim of gaining a better understanding of the internal burden of BHT in young nonspecifically exposed adults. The study population consisted of students between 20 and 29 years of age at the time of sampling, all from Halle/Saale in Central Germany. In total, 329 samples collected in the years 2000, 2004, 2008, 2012, 2015, and 2018 were measured by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). BHT acid was detected above the limit of quantification (0.2 My g/L) in 98% of the samples. The median of the measured concentrations was 1.06 My g/L and 1.24 My g/g creatinine respectively, the median of the daily excretion was 1.76 My g/24 h and - additionally normalized for body weight - 26.8 ng/24 h * kg bw respectively. The corresponding 90th percentiles were 3.28 My g/L, 3.91 My g/g creatinine, 5.05 My g/24 h, and 81.9 ng/24 h * kg bw. Medians of creatinine-corrected values were slightly higher in women than in men, while the opposite situation was observed for the volume concentrations and the 24-h excretion values (not corrected for body weight). Values simultaneously normalized both for 24-h excretion and body weight did not exhibit any significant differences between males and females, probably indicating a virtually identical magnitude of exposure for both genders. The background exposure of the investigated population was found to be largely constant since the year 2000, with only weak temporal trends at most. Daily intakes were estimated from excretion values and found to be largely below the acceptable daily intake (ADI) of BHT at 0.25 mg/kg bw: our worst-case estimate is a daily BHT intake of approximately 0.1 mg/kg bw at the 95th percentile level. However, these intake assessments rely on very limited quantitative data regarding human metabolism of BHT. © 2020 Elsevier GmbH. All rights reserved.

Comparison of software tools for kinetic evaluation of chemical degradation data

Background For evaluating the fate of xenobiotics in the environment, a variety of degradation or environmental metabolism experiments are routinely conducted. The data generated in such experiments are evaluated by optimizing the parameters of kinetic models in a way that the model simulation fits the data. No comparison of the main software tools currently in use has been published to date. This article shows a comparison of numerical results as well as an overall, somewhat subjective comparison based on a scoring system using a set of criteria. The scoring was separately performed for two types of uses. Uses of type I are routine evaluations involving standard kinetic models and up to three metabolites in a single compartment. Evaluations involving non-standard model components, more than three metabolites or more than a single compartment belong to use type II. For use type I, usability is most important, while the flexibility of the model definition is most important for use type II. Results Test datasets were assembled that can be used to compare the numerical results for different software tools. These datasets can also be used to ensure that no unintended or erroneous behaviour is introduced in newer versions. In the comparison of numerical results, good agreement between the parameter estimates was observed for datasets with up to three metabolites. For the now unmaintained reference software DegKinManager/ModelMaker, and for OpenModel which is still under development, user options were identified that should be taken care of in order to obtain results that are as reliable as possible. Based on the scoring system mentioned above, the software tools gmkin, KinGUII and CAKE received the best scores for use type I. Out of the 15 software packages compared with respect to use type II, again gmkin and KinGUII were the first two, followed by the script based tool mkin, which is the technical basis for gmkin, and by OpenModel. Conclusions Based on the evaluation using the system of criteria mentioned above and the comparison of numerical results for the suite of test datasets, the software tools gmkin, KinGUII and CAKE are recommended for use type I, and gmkin and KinGUII for use type II. For users that prefer to work with scripts instead of graphical user interfaces, mkin is recommended. For future software evaluations, it is recommended to include a measure for the total time that a typical user needs for a kinetic evaluation into the scoring scheme. It is the hope of the authors that the publication of test data, source code and overall rankings foster the evolution of useful and reliable software in the field. © The Author(s) 2018

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