A new class of contaminants of concern? A comprehensive review of liquid crystal monomers.

Liquid crystal monomers (LCMs) are a class of emerging contaminants of concern predicted to be persistent, bioaccumulative and toxic (PBT). Being one of the key components in liquid crystal displays (LCDs), the disposal of LCD containing devices is closely related to the emission of LCMs into the environment. LCMs have been detected in a wide range of environmental matrices including dust, sediment, soil, sewage leachate, and air, with concentration ranges between 17 and 2121 ng/g found in indoor residential dust. Furthermore, they have been detected on human skin at concentrations up to 2,071,000 ng/m2 and in the serum of e-waste dismantling workers, at concentrations ranging from 3.9 to 276 ng/mL. Despite the far-reaching contamination of these compounds, there is limited knowledge of their environmental behaviour, fate, and toxicity. Model predictions show that 297 of 330 LCMs are persistent and bioaccumulative compounds, with many more indicated as being toxic. However, current knowledge of their physicochemical and PBT properties is largely restricted to theoretical predictions and limited to a small number of experimental toxicity studies. As an emerging class of contaminants of concern, a lack of standardisation between studies was identified as a key challenge to advancing the state of knowledge of these compounds. Not only are harmonised analytical methods for their determination and quantification in environmental media yet to be established, but there is also a need for a universal abbreviation system. To further harmonise the reporting of data on LCMs we propose reporting the sum concentration of ten priority LCMs, selected on the basis detection frequency, toxicity and potential for human exposure. Of the ten priority LCMs five are fluorinated biphenyls and analogues, four are biphenyls/bicyclohexyls and analogues and one is a cyanobiphenyl.

In vitro screening of understudied PFAS with a focus on lipid metabolism disruption.

Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals used in many industrial applications. Exposure to PFAS is associated with several health risks, including a decrease in infant birth weight, hepatoxicity, disruption of lipid metabolism, and decreased immune response. We used the in vitro cell models to screen six less studied PFAS [perfluorooctane sulfonamide (PFOSA), perfluoropentanoic acid (PFPeA), perfluoropropionic acid (PFPrA), 6:2 fluorotelomer alcohol (6:2 FTOH), 6:2 fluorotelomer sulfonic acid (6:2 FTSA), and 8:2 fluorotelomer sulfonic acid (8:2 FTSA)] for their capacity to activate nuclear receptors and to cause differential expression of genes involved in lipid metabolism. Cytotoxicity assays were run in parallel to exclude that observed differential gene expression was due to cytotoxicity. Based on the cytotoxicity assays and gene expression studies, PFOSA was shown to be more potent than other tested PFAS. PFOSA decreased the gene expression of crucial genes involved in bile acid synthesis and detoxification, cholesterol synthesis, bile acid and cholesterol transport, and lipid metabolism regulation. Except for 6:2 FTOH and 8:2 FTSA, all tested PFAS downregulated PPARA gene expression. The reporter gene assay also showed that 8:2 FTSA transactivated the farnesoid X receptor (FXR). Based on this study, PFOSA, 6:2 FTSA, and 8:2 FTSA were prioritized for further studies to confirm and understand their possible effects on hepatic lipid metabolism.

Legacy and alternative per- and polyfluoroalkyl substances (PFAS) alter the lipid profile of HepaRG cells.

Per- and polyfluoroalkyl substances (PFAS) are synthetic chemicals used in various industrial and consumer products. They have gained attention due to their ubiquitous occurrence in the environment and potential for adverse effects on human health, often linked to immune suppression, hepatotoxicity, and altered cholesterol metabolism. This study aimed to explore the impact of ten individual PFAS, 3 H-perfluoro-3-[(3-methoxypropoxy) propanoic acid] (PMPP/Adona), ammonium perfluoro-(2-methyl-3-oxahexanoate) (HFPO-DA/GenX), perfluorobutanoic acid (PFBA), perfluorobutanesulfonic acid (PFBS), perfluorodecanoic acid (PFDA), perfluorohexanoic acid (PFHxA), perfluorohexanesulfonate (PFHxS), perfluorononanoic acid (PFNA), perfluorooctanoic acid (PFOA), and perfluorooctanesulfonic acid (PFOS) on the lipid metabolism in human hepatocyte-like cells (HepaRG). These cells were exposed to different concentrations of PFAS ranging from 10 µM to 5000 µM. Lipids were extracted and analyzed using liquid chromatography coupled with mass spectrometry (LC- MS-QTOF). PFOS at 10 µM and PFOA at 25 µM increased the levels of ceramide (Cer), diacylglycerol (DAG), N-acylethanolamine (NAE), phosphatidylcholine (PC), and triacylglycerol (TAG) lipids, while PMPP/Adona, HFPO-DA/GenX, PFBA, PFBS, PFHxA, and PFHxS decreased the levels of these lipids. Furthermore, PFOA and PFOS markedly reduced the levels of palmitic acid (FA 16.0). The present study shows distinct concentration-dependent effects of PFAS on various lipid species, shedding light on the implications of PFAS for essential cellular functions. Our study revealed that the investigated legacy PFAS (PFOS, PFOA, PFBA, PFDA, PFHxA, PFHxS, and PFNA) and alternative PFAS (PMPP/Adona, HFPO-DA/GenX and PFBS) can potentially disrupt lipid homeostasis and metabolism in hepatic cells. This research offers a comprehensive insight into the impacts of legacy and alternative PFAS on lipid composition in HepaRG cells.

The profile of steroid hormones in human fetal and adult ovaries.

BACKGROUND: Reproduction in women is at risk due to exposure to chemicals that can disrupt the endocrine system during different windows of sensitivity throughout life. Steroid hormone levels are fundamental for the normal development and function of the human reproductive system, including the ovary. This study aims to elucidate steroidogenesis at different life-stages in human ovaries. METHODS: We have developed a sensitive and specific LC-MS/MS method for 21 important steroid hormones and measured them at different life stages: in media from cultures of human fetal ovaries collected from elective terminations of normally progressing pregnancy and in media from adult ovaries from Caesarean section patients, and follicular fluid from women undergoing infertility treatment. Statistically significant differences in steroid hormone levels and their ratios were calculated with parametric tests. Principal component analysis (PCA) was applied to explore clustering of the ovarian-derived steroidogenic profiles. RESULTS: Comparison of the 21 steroid hormones revealed clear differences between the various ovarian-derived steroid profiles. Interestingly, we found biosynthesis of both canonical and "backdoor" pathway steroid hormones and corticosteroids in first and second trimester fetal and adult ovarian tissue cultures. 17α-estradiol, a less potent naturally occurring isomer of 17ß-estradiol, was detected only in follicular fluid. PCA of the ovarian-derived profiles revealed clusters from: adult ovarian tissue cultures with relatively high levels of androgens; first trimester and second trimester fetal ovarian tissue cultures with relatively low estrogen levels; follicular fluid with the lowest androgens, but highest corticosteroid, progestogen and estradiol levels. Furthermore, ratios of specific steroid hormones showed higher estradiol/ testosterone and estrone/androstenedione (indicating higher CYP19A1 activity, p < 0.01) and higher 17-hydroxyprogesterone/progesterone and dehydroepiandrosterone /androstenedione (indicating higher CYP17A1 activity, p < 0.01) in fetal compared to adult ovarian tissue cultures. CONCLUSIONS: Human ovaries demonstrate de novo synthesis of non-canonical and "backdoor" pathway steroid hormones and corticosteroids. Elucidating the steroid profiles in human ovaries improves our understanding of physiological, life-stage dependent, steroidogenic capacity of ovaries and will inform mechanistic studies to identify endocrine disrupting chemicals that affect female reproduction.

Exploring the Relationship Among Lipid Profile Changes, Growth, and Reproduction in Folsomia candida Exposed to Teflubenzuron Over Time.

The integration of untargeted lipidomics approaches in ecotoxicology has emerged as a strategy to enhance the comprehensiveness of environmental risk assessment. Although current toxicity tests with soil microarthropods focus on species performance, that is, growth, reproduction, and survival, understanding the mechanisms of toxicity across all levels of biological organization, from molecule to community is essential for informed decision-making. Our study focused on the impacts of sublethal concentrations of the insecticide teflubenzuron on the springtail Folsomia candida. Untargeted lipidomics was applied to link changes in growth, reproduction, and the overall stress response with lipid profile changes over various exposure durations. The accumulation of teflubenzuron in organisms exposed to the highest test concentration (0.035 mg a.s. kg-1 soil dry wt) significantly impacted reproductive output without compromising growth. The results suggested a resource allocation shift from reproduction to size maintenance. This hypothesis was supported by lipid shifts on day 7, at which point reductions in triacylglycerol and diacylglycerol content corresponded with decreased offspring production on day 21. The hypermetabolism of fatty acids and N-acylethanolamines on days 2 and 7 of exposure indicated oxidative stress and inflammation in the animals in response to teflubenzuron bioaccumulation, as measured using high-performance liquid chromatography-tandem mass spectrometry. Overall, the changes in lipid profiles in comparison with phenotypic adverse outcomes highlight the potential of lipid analysis as an early-warning tool for reproductive disturbances caused by pesticides in F. candida. Environ Toxicol Chem 2024;43:1149-1160. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

A Rapid Screening Method for the Detection of Additives in Electronics and Plastic Consumer Products Using AP-MALDI-qTOF-MS.

A novel method was developed and optimized for the fast-screening analysis of additives in electronics and plastic consumer products using atmospheric pressure matrix-assisted laser desorption ionization (AP-MALDI) coupled with a high-resolution quadrupole time-of-flight (qTOF) mass spectrometer (MS). To simplify sample preparation and increase sample throughput, an innovative 48 well graphene nanoplatelets (GNP) doped AP-MALDI target plate was developed. The GNP incorporated in the target plate fulfilled the role of the MALDI matrix and, therefore, sample extracts could be directly transferred to the AP-MALDI 48 well target plate and analyzed without a subsequent matrix addition. The homogeneously dispersed and immobilized GNP target plates also provided increased signal intensity and reproducibility. Furthermore, analytical standards of various plastic additives and plastic products with known concentrations of additives were studied to assess the AP-MALDI ionization mechanisms and method capability. The analysis time was 15 s per measurement using an automated sequence. The GNP-doped target plates exhibited high desorption/ionization of low molecular weight molecules (<1000 Da) and can be used in both positive and negative ionization modes. The AP-MALDI-qTOF-MS method was applied to screen for additives in various electronics and plastic consumer products. Suspect screening was performed using a database containing 1366 compounds. A total of 56 additives including antioxidants, flame retardants, plasticizers, UV-stabilizers, and UV-filters were identified (confidence level 4). Identification of certain plastic additives in plastic children's toys may indicate that they are recycled from waste electronic and electronic equipment (WEEE).

Identification of biomarkers and outcomes of endocrine disruption in human ovarian cortex using In Vitro Models.

Endocrine disrupting chemicals (EDCs) are raising concerns about adverse effects on fertility in women. However, there is a lack of information regarding mechanisms and effects in humans. Our study aims to identify mechanisms of endocrine disruption using two EDCs, diethylstilbestrol (DES) and ketoconazole (KTZ)1. Human ovarian cortical tissue obtained from Caesarean section patients was exposed to 10-9 M - 10-5 M KTZ and 10-10 M - 10-6 M DES in vitro for 6 days. Follicle survival and growth were studied via histology analysis and liquid-chromatography-mass spectrometry-based steroid quantification. RNA-sequencing was performed on COV434, KGN, and primary ovarian cells that were exposed for 24 h. Significantly lower unilaminar follicle densities were observed in DES 10-10 M group, whereas low KTZ exposure reduced secondary follicle density. KTZ 10-5 M reduced levels of pregnenolone and progesterone. RNA-sequencing revealed that 445 and 233 differentially expressed genes (false discovery rate < 0.1) altogether in DES and KTZ exposed groups. Gene set variation analysis showed that both chemicals modulated pathways that are important for folliculogenesis and steroidogenesis. We selected stearoyl-CoA desaturase (SCD) and 7-dehydrocholesterol reductase (DHCR7) for further validation. Up-regulation of both genes in response to KTZ was confirmed by qPCR and in situ RNA hybridization. Further validation with immunofluorescence focused on the expression of SCD in growing follicles in exposed ovarian tissue. In conclusion, SCD may serve as a potential novel human-relevant biomarker of EDC exposure and effects on ovaries.

Metabolite alterations in zebrafish embryos exposed to hydroxylated polybrominated diphenyl ethers.

Hydroxylated polybrominated diphenyl ethers (OH-PBDEs) are formed by metabolism from the flame retardants polybrominated diphenyl ethers (PBDEs). In the aquatic environment, they are also produced naturally. OH-PBDEs are known for their potential to disrupt energy metabolism, the endocrine system, and the nervous system. This is the first study focusing on the effects of OH-PBDEs at the metabolite level in vivo. The aim of the current study was to investigate the metabolic effects of exposure to OH-PBDEs using metabolomics, and to identify potential biomarker(s) for energy disruption of OH-PBDEs. Zebrafish (Danio rerio) embryos were exposed to two different concentrations of 6-OH-BDE47 and 6-OH-BDE85 and a mixture of these two compounds. In total, 342 metabolites were annotated and 79 metabolites were affected in at least one exposure. Several affected metabolites, e.g. succinic acid, glutamic acid, glutamine, tyrosine, tryptophan, adenine, and several fatty acids, could be connected to known toxic mechanisms of OH-PBDEs. Several phospholipids were strongly up-regulated with up to a six-fold increase after exposure to 6-OH-BDE47, a scarcely described effect of OH-PBDEs. Based on the observed metabolic effects, a possible connection between disruption of the energy metabolism, neurotoxicity and potential immunotoxicity of OH-PBDEs was suggested. Single compound exposures to 6-OH-BDE47 and 6-OH-BDE85 showed little overlap in the affected metabolites. This shows that compounds of similar chemical structure can induce different metabolic effects, possibly relating to their different toxic mechanisms. There were inter-concentration differences in the metabolic profiles, indicating that the metabolic effects were concentration dependent. After exposure to the mixture of 6-OH-BDE47 and 6-OH-BDE85, a new metabolic profile distinct from the profiles obtained from the single compounds was observed. Succinic acid was up-regulated at the highest, but still environmentally relevant, concentration of 6-OH-BDE47, 6-OH-BDE85, and the mixture. Therefore, succinic acid is suggested as a potential biomarker for energy disruption of OH-PBDEs.

Fate of Per- and Polyfluoroalkyl Substances from Durable Water-Repellent Clothing during Use.

To make outdoor clothing water- or dirt-repellent, durable water-repellent (DWR) coatings based on side-chain fluorinated polymers (SFPs) are used. During use of outdoor clothing, per- and polyfluoroalkyl substances (PFASs) can be emitted from the DWR to the environment. In this study, the effects of aging, washing, and tumble drying on the concentration of extractable PFASs in the DWR of perfluorohexane-based short-chain SFPs (FC-6 chemistry) and of perfluorooctane-based long-chain SFPs (FC-8 chemistry) were assessed. For this purpose, polyamide (PA) and polyester (PES) fabrics were coated with FC-6- and FC-8-based DWRs. Results show that aging of the coated fabrics causes an increase in concentration and formation of perfluoroalkyl acids (PFAAs). The effect of aging on the volatile PFASs depends on the type of fabric. Washing causes a decrease in PFAA concentrations, and in general, volatile PFASs are partly washed out of the textiles. However, washing can also increase the extractable concentration of volatile PFASs in the fabrics. This effect becomes stronger by a combination of aging and washing. Tumble drying does not affect the PFAS concentrations in textiles. In conclusion, aging and washing of fabrics coated with the DWR based on SFPs release PFASs to the environment.

Migration of hazardous contaminants from WEEE contaminated polymeric toy material by mouthing.

This study evaluated the migration of brominated flame retardants (BFRs), phosphate flame retardants (PFRs), bisphenols (BPA, BPF), and phthalate ester-based plasticizers from recycled polymeric toy material, containing waste electrical and electronic equipment (WEEE), in artificial saliva simulating 1 h of mouthing. In total 12 parts of 9 different toys were tested in triplicate after confirming WEEE specific contamination. Up to 11 contaminants were detected in saliva from one toy sample. The highest migration rate up to 128 ng/(cm2 x h) was found for BPA followed by bis(2-ethylhexyl) phthalate (DEHP) and diisobutyl phthalate (DIBP) with migration rates up to 25.5 and 8.27 ng/(cm2 x h), respectively. In addition to DecaBDE, which was detected in 3 saliva samples at migration rates between 0.09 and 0.31 ng/(cm2 x h), the decaBDE replacements 2,4,6-tris(2,4,6-tribromophenoxy)-1,3,5-triazine (TTBP-TAZ), decabromodiphenyl ethane (DBDPE), resorcinol bis(diphenyl phosphate) (RDP), and hexabromocyclododecane (HBCDD) were detected as well with comparable migration rates. 2,4,6-tribromphenol (246-TBP) reached migration rates up to 1.15 ng/(cm2 x h) in correspondence to the presence of TTBP-TAZ. Tetrabromobisphenol A (TBBPA), BPA, 246-TBP, DEHP, DIBP and triphenyl phosphate (TPHP) were predominantly observed in saliva with a detection frequency between 50 and 75%. Daily intake (DI) values were calculated for relevant analytes and compared to tolerable daily intake (TDI) values. The highest DI values of 72.4, 14.3, 5.74, 2.28 and 2.09 ng/(kg BW x day), were obtained for BPA, DEHP, DIBP, TBBPA, and TPHP, respectively. None of them exceed the TDI value or respective reference dose (RfD).

Chlorinated paraffins and tris (1-chloro-2-propyl) phosphate in spray polyurethane foams - A source for indoor exposure?

In this study, we investigated chemical additives present in new and used spray polyurethane foams (SPFs) and assessed the dermal transfer through direct contact. This first study shows that cured do-it-yourself spray one-component SPFs (OCFs) often contain chlorinated paraffins (C14-C37), and tris (1-chloro-2-propyl) phosphate (TCIPP), ranging 0.2-50%, and 0.9-30% w/w, respectively. Six OCFs contained CP levels ranging 22-50% w/w, whereas nine OCFs used for similar applications only contained CP levels ranging 2-17% w/w. It is unclear if the combination CPs/TCIPP is meant to improve the flame retardancy of products, and could suggest an unnecessary use of high CPs/TCIPP concentrations in OCFs. The two-component SPFs (TCFs) contained only TCIPP with levels ranging from 7.0% to 9.0%. The CPs and TCIPP were easily transferred from cured OCFs to the hands. Levels up to 590 µg per hand for CPs and up to 2.7 µg per hand for TCIPP were found. After end-of-life, it is challenging to recycle used SPFs. They may, therefore, end up at landfills where the TCIPP/CPs may leach into the environment. Therefore, further investigation is needed to assess potential exposure risks associated with general and occupational use, and the impact of landfill leaching on the environment.

Progress towards an OECD reporting framework for transcriptomics and metabolomics in regulatory toxicology.

Omics methodologies are widely used in toxicological research to understand modes and mechanisms of toxicity. Increasingly, these methodologies are being applied to questions of regulatory interest such as molecular point-of-departure derivation and chemical grouping/read-across. Despite its value, widespread regulatory acceptance of omics data has not yet occurred. Barriers to the routine application of omics data in regulatory decision making have been: 1) lack of transparency for data processing methods used to convert raw data into an interpretable list of observations; and 2) lack of standardization in reporting to ensure that omics data, associated metadata and the methodologies used to generate results are available for review by stakeholders, including regulators. Thus, in 2017, the Organisation for Economic Co-operation and Development (OECD) Extended Advisory Group on Molecular Screening and Toxicogenomics (EAGMST) launched a project to develop guidance for the reporting of omics data aimed at fostering further regulatory use. Here, we report on the ongoing development of the first formal reporting framework describing the processing and analysis of both transcriptomic and metabolomic data for regulatory toxicology. We introduce the modular structure, content, harmonization and strategy for trialling this reporting framework prior to its publication by the OECD.

Combining High-Resolution Gas Chromatographic Continuous Fraction Collection with Nuclear Magnetic Resonance Spectroscopy: Possibilities of Analyzing a Whole GC Chromatogram.

This study presents, for the first time, the successful application of analyzing a whole gas chromatography (GC) chromatogram by nuclear magnetic resonance (NMR) spectroscopy using a continuous repeatable and stable (n = 280) high-resolution (HR) GC fractionation platform with a 96-well plate. Typically with GC- or liquid chromatography-mass spectrometry analysis, (isomer) standards and/or additional NMR analysis are needed to confirm the identification and/or structure of the analyte of interest. In the case of complex substances (e.g., UVCBs), isomer standards are often unavailable and NMR spectra too complex to achieve this. This proof of concept study shows that a HR GC fractionation collection platform was successfully applied to separate, purify, and enrich isomers in complex substances from a whole GC chromatogram, which would facilitate NMR analysis. As a model substance, a chlorinated paraffin (CP) mixture (>8,000 isomers) was chosen. NMR spectra were obtained from all 96 collected fractions, which provides important information for unravelling their full structure. As a proof of concept, a spectral interpretation of a few NMR spectra was made to assign sub-structures. More research is ongoing for the full characterization of CP isomers using multivariate statistical analysis. For the first time, up to only a few CP isomers per fraction were isolated from a highly complex mixture. These may be further purified and certified as standards, which are urgently needed, and can also be used for persistency, bioaccumulation, or toxicity studies.

Analysis of recycled rubber: Development of an analytical method and determination of polycyclic aromatic hydrocarbons and heterocyclic aromatic compounds in rubber matrices.

Recycled crumb rubber (CR) is rich in compounds with unrecognized toxic potency; this study aims at the development of an analytical method that would allow identification and quantification of a very wide range of organic compounds extractable from the complex rubber matrix. The analytical set-up includes target analysis of polycyclic aromatic hydrocarbons (PAHs) and methyl-PAHs and suspect screening of raw extracts to tentatively identify primary organic compounds present, but not included in the standard target analysis of recycled rubber, followed by analytical method development and target analysis of identified compounds. Analyzed samples included weathered and new CR originating from football turf granulates, rubber mats, and end-of-life car tires (ELTs). The developed analytical method involves sonication extraction, followed by solid phase extraction (SPE) fractionation that enables simple and efficient separation of analytes with broad polarity scale. The application of SPE fractionation resolves coelution problems and simplifies the chromatograms. This analytical approach allowed to identify and quantify 46 sample specific compounds, including several heterocyclic PAHs like 2-methylthiobenzothiazole, benzonapthothiophenes, benzonaphthofuranes and aromatic amines like diphenylamine and N-phenyl-2-naphthylamine, which to our knowledge were not determined before. The PAHs concentrations determined in CR tiles purchased in Dutch and Spanish shops exceed the EU limits for articles marketed for use by the public. Furthermore, sets of methylated PAHs, dibenzothiazoles and aromatic amines were identified and quantified, and several other compounds were tentatively identified. The obtained results stress the need for expanding the list of target compounds analyzed in CR and the need for longitudinal studies on weathering processes taking place in CR.

In vitro biotransformation and evaluation of potential transformation products of chlorinated paraffins by high resolution accurate mass spectrometry.

Chlorinated paraffins (CPs) are high production chemicals, which leads to their ubiquitous presence in the environment. To date, few studies have measured CPs in humans and typically at relatively low concentrations, despite indications that exposure may be high compared to various persistent organic pollutants. The aim of this study is to investigate the in vitro biotransformation of CPs by human liver fractions. We determined the changes of the CP concentrations after the enzymatic transformation with human liver microsomes using a two-tiered in vitro approach. CP concentrations decreased with human liver microsomes, with the decreases of 33-94% after incubating with different groups of enzymes for 2 h. The profiles of CP rapidly shifted after the incubation with human liver microsomes. In addition, the concentrations of CPs and the biotransformation products were tentatively measured using high-resolution mass spectrometric analysis, including very short CP (carbon chain length <10), alcohols, ketones, and carboxylic acids. C‒C bond cleavage is a potential transformation pathway for CPs, and ketones are potential products of CP biotransformation, especially for long-chain CPs (C>17). The ketone products may be investigated as CP exposure biomarker in biomonitoring studies.

The ENDpoiNTs Project: Novel Testing Strategies for Endocrine Disruptors Linked to Developmental Neurotoxicity.

Ubiquitous exposure to endocrine-disrupting chemicals (EDCs) has caused serious concerns about the ability of these chemicals to affect neurodevelopment, among others. Since endocrine disruption (ED)-induced developmental neurotoxicity (DNT) is hardly covered by the chemical testing tools that are currently in regulatory use, the Horizon 2020 research and innovation action ENDpoiNTs has been launched to fill the scientific and methodological gaps related to the assessment of this type of chemical toxicity. The ENDpoiNTs project will generate new knowledge about ED-induced DNT and aims to develop and improve in vitro, in vivo, and in silico models pertaining to ED-linked DNT outcomes for chemical testing. This will be achieved by establishing correlative and causal links between known and novel neurodevelopmental endpoints and endocrine pathways through integration of molecular, cellular, and organismal data from in vitro and in vivo models. Based on this knowledge, the project aims to provide adverse outcome pathways (AOPs) for ED-induced DNT and to develop and integrate new testing tools with high relevance for human health into European and international regulatory frameworks.

Toddler behavior, the home environment, and flame retardant exposure.

Toddlers are at increased risk of dust ingestion and subsequently flame retardant (FR) exposure because they often play close to the floor and mouth hands and objects. Exposure to some FRs have been associated to endocrine disruption and neurodevelopmental disorders. Previous research has shown higher FR concentrations in toddlers' serum and urine, but which toddler-behaviors influence exposure levels remains to be determined. We investigated how toddler-behaviors are associated to FRs in hand wipes (HWs) and saliva. Fifty 8-18 month-old children from the Linking EDCs in maternal Nutrition to Child health study, were visited at home. The child's behavior was observed and assessed using a questionnaire. Hand-to-object behavior frequency was associated with HW tris(chloroethyl) phosphate (TCEP), tris(1,3-dichloroisopropyl) phosphate (TDCIPP), tris(phenyl) phosphate, tris(methylphenyl) phosphate, and resorcinol bis(diphenyl phosphate) levels above the detection limit. Children playing with electronics multiple times per week had higher TDCIPP HW levels compared to children playing with electronics once per month or never (p = 0.032 and p = 0.046). Frequent mouth-to-object and frequent mouthing a pacifier were associated with lower TDCIPP (p = 0.019) and tris(2-chloroisopropyl) phosphate (TCIPP) HW levels, respectively (p = 0.002-0.019). Exposure estimates based on hand-to-mouth behavior did not exceed the available reference doses. This is the first study investigating toddler-behavior in relation to FR hand loadings. Although a range of behaviors was investigated, only a few showed a relation with FR HW levels, suggesting that toddler-behavior might not alone be responsible for the elevated FR levels in children. It is therefore important to explore other pathways including dermal absorption and inhalation.

Spatial variation of short- and medium-chain chlorinated paraffins in ambient air across Australia.

Atmospheric levels of chlorinated paraffins (CPs) at five remote, six rural and four urban sites in Australia were measured using XAD-2 passive air samplers (XAD-PAS). While long-chain CP (LCCP, C>17) levels were below method detection limits (MDLs), short-chain CPs (SCCPs, C10-13) and, for the first time, medium-chain CPs (MCCPs, C14-17) and CPs with a carbon chain length of nine (CP-C9) were found at many sites (88%, 81% and 88%, respectively) across the Australian continent, representing a range of environmental conditions. Applying preliminary sampling rates of the XAD-PAS for CPs, gaseous CP levels in Australian air were

The effect of weathering on per- and polyfluoroalkyl substances (PFASs) from durable water repellent (DWR) clothing.

To assess the effects of weathering on per- and polyfluoroalkyl substances (PFASs) from durable water repellent (DWR) clothing, thirteen commercial textile samples were exposed to elevated ultra violet (UV) radiation, humidity, and temperature in an aging device for 300 h, which mimics the lifespan of outdoor clothing. Before and after aging, the textile samples were extracted and analysed for the ionic PFASs (perfluoroalkyl acids (PFAAs), perfluorooctane sulfonamide (FOSA)) and volatile PFASs (fluorotelomer alcohols (FTOHs), acrylates (FTACs) and methacrylates (FTMACs)). Results showed that weathering can have an effect on PFASs used in DWR of outdoor clothing, both on the PFAS profile and on the measured concentrations. In most weathered samples the PFAA concentrations increased by 5- to more than 100-fold, while PFAAs not detected in the original textiles were detected in the weathered samples. DWR chemistries are based on side-chain fluorinated polymers. A possible explanation for the increase in concentration of the PFAAs is hydrolysis of the fluorotelomer based polymers (FTPs), or degradation of the FTOHs, which are used in the manufacturing of the FTPs. The concentrations of volatile PFASs also increased, by a factor up to 20. Suggested explanations are the degradation of the DWR polymers, making non-extractable fluorines extractable, or the transformation or degradation of unknown precursors. Further research is needed to unravel the details of these processes and to determine the transformation routes. This study shows that setting maximum tolerance limits only for a few individual PFASs is not sufficient to control these harmful substances in outdoor clothing.