Phthalate and DINCH metabolites in the urine of Hungarian schoolchildren: Cumulative risk assessment and exposure determinants.

A human biomonitoring study was conducted to assess the exposure of Hungarian children aged 8-11 years to ten phthalate esters (PEs) and DINCH between 2017 and 2018. In addition to collecting urine samples from 262 participants, a questionnaire was completed by the parents or legal guardians to identify potential determinants of exposure. The highest geometric mean concentration was observed for MiBP, followed by MBP, cx- MEHP, OH-MEHP and MEP. Three out of the four DINCH metabolites were detected in more than 90% of the samples. The comparison of the urinary concentrations measured in this study with those observed in the DEMOCOPHES study revealed a significant decreasing trend in all PE metabolites investigated in both studies between 2011/2012 and 2017/2018. Different approaches were used to assess the health risks associated with the exposure to PEs and DINCH. Our results highlighted that the hazard index (HI) values were higher than 1 in 17.6% of the children when the human biomonitoring guidance values were applied. In contrast, less than 3% of the children had HI values exceeding 1 when other sources of reference values were used. By applying a safety factor of 10 for the risk assessment, 17.6-91.6% of the children were characterized by HI values higher than 0.1, indicating the need for risk reduction measures. Overall, DnBP, DiBP and DEHP were identified as the main drivers of the mixture risk. Although PEs and DINCH are ubiquitous contaminants, there are still inconsistencies and gaps in our understanding of the determinants of exposure. The results of the multivariate regression analysis showed significant associations between PE or DINCH metabolite concentrations and certain individual characteristics, use of personal care products, home and school environment and food and beverages consumption 24 h prior to sample collection.

Prenatal exposure to polycyclic aromatic hydrocarbons and growth parameters.

BACKGROUND AND OBJECTIVES: It has long been known that airborne polycyclic aromatic hydrocarbons (PAHs) can negatively affect pregnancy and birth outcomes, such as birth weight, fetal development, and placental growth factors. However, similar studies yield divergent results. Our goal was to estimate the amount of monohydroxylated PAH (OH-PAH) metabolites in the urine of pregnant women/mothers and their newborns in relation to birth outcomes, such as placenta weight, Apgar 5', and the growth parameters of children up to the age of two. METHODS: Two cohorts of children born in 2013 and 2014 during the summer and winter seasons in the Czech Republic in the cities Karviná (N = 144) and Ceské Budejovice (N = 198), which differ significantly in the level of air pollution, were studied. PAH exposure was assessed by the concentration of benzo[a]pyrene (B[a]P) in the air and the concentration of 11 OH-PAH metabolites in the urine of newborns and mothers. Growth parameters and birth outcomes were obtained from medical questionnaires after birth and from pediatric questionnaires during the following 24 months of the child's life. RESULTS: Concentrations of B[a]P were significantly higher in Karviná (p < 0.001). OH-PAH metabolites were significantly higher in the mothers' as well as in the newborns' urine in Karviná and during the winter season. Neonatal length was shorter in newborns in Karviná (p < 0.001), but this difference evened out during the next 3 to 24 months. Compared to Ceské Budejovice, newborns in Karviná showed significantly lower weight gain between birth and three months after delivery. The OH-PAH metabolites in mothers' or newborns' urine did not affect birth weight. The presence of seven OH-PAH (top 25% of values of concentrations higher than the median) metabolites in the newborns' urine is associated with decreased length of newborn. Nine OH-PAH metabolites decreased placenta weight, which was the most significant, while seven OH-PAH metabolites decreased Apgar 5'. CONCLUSION: We have shown a possible connection between higher concentration of OH-PAH metabolites in newborns' urine and decreased length, head circumference, placenta weight, and Apgar 5', but not birth weight.

European interlaboratory comparison investigations (ICI) and external quality assurance schemes (EQUAS) for the analysis of bisphenol A, S and F in human urine: Results from the HBM4EU project.

The Human Biomonitoring for Europe initiative (HBM4EU) aims to study the exposure of citizens to chemicals and potentially associated health effects. One objective of this project has been to build a network of laboratories able to answer to the requirements of European human biomonitoring studies. Within the HBM4EU quality assurance and quality control scheme (QA/QC), a number of interlaboratory comparison investigations (ICIs) and external quality assurance schemes (EQUASs) were organized to ensure data consistency, comparability and reliability. Bisphenols are among the prioritized substance groups in HBM4EU, including bisphenol A (BPA), bisphenol S (BPS) and bisphenol F (BPF) in human urine. In four rounds of ICI/EQUAS, two target concentration levels were considered, related to around P25 and P95 of the typical exposure distribution observed in the European general population. Special attention was paid to the conjugated phase II metabolites known to be most dominant in samples of environmentally exposed individuals, through the analysis of both native samples and samples fortified with glucuronide forms. For the low level, the average percentage of satisfactory results across the four rounds was 83% for BPA, 71% for BPS and 62% for BPF. For the high level, the percentages of satisfactory results increased to 93% for BPA, 89% for BPS and 86% for BPF. 24 out of 32 participating laboratories (75%) were approved for the analyses of BPA in the HBM4EU project according to the defined criterion of Z-scores for both low and high concentration levels in at least two ICI/EQUAS rounds. For BPS and BPF, the number of qualified laboratories was 18 out of 27 (67%) and 13 out of 28 (46%), respectively. These results demonstrate a strong analytical capability for BPA and BPS in Europe, while improvements may be needed for BPF.

Interlaboratory comparison investigations (ICIs) and external quality assurance schemes (EQUASs) for flame retardant analysis in biological matrices: Results from the HBM4EU project.

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 analysing flame retardants (FRs) in serum and urine were investigated by a quality assurance/quality control (QA/QC) scheme 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 performed from 2018 to 2020 for the determination of ten halogenated flame retardants (HFRs) represented by three congeners of polybrominated diphenyl ethers (BDE-47, BDE-153 and BDE-209), two isomers of 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.

Biomonitoring of PFOA, PFOS and PFNA in human milk from Czech Republic, time trends and estimation of infant's daily intake.

BACKGROUND: Perfluoralkylated substances (PFASs) are persistent and bioaccumulative environmental contaminants. They are included on the list of emergent compounds monitored in the frame of HBM4EU project. OBJECTIVES: To analyze PFASs levels in human milk samples collected in the period 2006 through 2017, to follow their time trends, to assess the PFASs exposure in breastfed infants, to calculate the daily intake of PFASs and to compare it with the tolerable daily/weekly) intakes and to quantify risk from exposure using the hazard quotient and hazard index approach. MATERIAL AND METHODS: A broad spectrum of PFASs were analyzed by means of UHPLC-MS/MS in primipara human milk samples collected in four consecutive time periods 2006, 2010/11, 2014, and 2017; N = 46, 183, 164 and 232, respectively. Mothers living in urban and suburban residences were recruited after their delivery at maternity hospitals, and milk samples were taken within 2 and 8 weeks after delivery. The questionnaire was focused on possible sources of exposure, dietary habits and lifestyle. RESULTS: Only perfluorooctane sulfonate (PFOS) and perfluorooctanoid acid PFOA (in 2017, also perfluorononanoic acid (PFNA)) were quantified in more than 90% of analyzed human milk samples. In all sampling periods, the levels of PFOA were higher than those of PFOS (p < 0.05). A significant downward temporal trend (p < 0.001) was observed for both PFOA and PFOS levels. The median concentrations in sampling years 2006, 2010/11, 2014, and 2017 were 0.075, 0.059, 0.035, and 0.023 ng/mL for PFOA and 0.045, 0.031, 0.029, and 0.020 ng/mL for PFOS, respectively. In 2017, PFNA was also quantified in 99% of samples with the median concentration of 0.007 ng/mL. The levels of PFASs correlated with maternal sea fish consumption. No maternal age-related relationship was observed. Using the tolerably daily intake (TDI) values for PFOS and PFOA set by the European Food Safety Authority (EFSA) in 2008, the calculated daily intakes from breastfeeding were clearly below these limits. Using the new, more conservative EFSA Provisional Tolerably Weekly Intake (PTWI) values set in 2018, we demonstrated a considerable exceedance of PTWI, with a hazard index above 1. CONCLUSION: Significant time-related decreasing trends in the PFOS and PFOA levels in human milk were observed. Nevertheless, the body burden of infants from breastfeeding might pose an enhanced health risk to infants when the current PTWI values are applied. These findings strongly support the present EU efforts to phase out PFOA, its salts and PFOA related compounds. Since PFOS exposure there has still been widely detected despite PFOS usage reduction measures, the major exposure routes should be further monitored and, if possible, eliminated.

Bisphenols in daily clothes from conventional and recycled material: evaluation of dermal exposure to potentially toxic substances.

Given the increasing concern about chemical exposure from textiles, our study examines the risks of dermal exposure to bisphenol A (BPA), bisphenol S (BPS), bisphenol B (BPB) and bisphenol F (BPF) from conventional and recycled textiles for adults, aiming to obtain new data, assess exposure, and evaluate the impact of washing on bisphenol levels. A total of 57 textile samples (33 from recycled and 24 from conventional material) were subjected to ultrasound-assisted extraction (UAE) followed by ultra-high performance liquid chromatography with tandem mass spectrometry analysis (UHPLC-MS/MS). The BPA and BPS concentrations varied widely (BPA: < 0.050 to 625 ng/g, BPS: 0.277-2,474 ng/g). The median BPA content in recycled textiles (13.5 ng/g) was almost twice as high as that of 7.66 ng/g in conventional textiles. BPS showed a median of 1.85 ng/g in recycled textiles and 3.42 ng/g in conventional textiles, indicating a shift from BPA to BPS in manufacturing practices. Simulated laundry experiments showed an overall reduction in bisphenols concentrations after washing. The study also assessed potential health implications via dermal exposure to dry and sweat-wet textiles compared to a tolerable daily intake (TDI) of 0.2 ng/kg bw/day for BPA set by the European Food Safety Authority (EFSA). Exposure from dry textiles remained below this threshold, while exposure from wet textiles often exceeded it, indicating an increased risk under conditions that simulate sweating or humidity. By finding the widespread presence of bisphenols in textiles, our study emphasises the importance of being aware of the potential risks associated with recycling materials as well as the benefits.

Novel strategies for the determination of plastic additives derived from agricultural plastics in soil using ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS).

Certain agricultural plastics, i.e., mulching films, are generally considered as potent sources of micro- and nanoplastics (MNPs), due to their direct application on soil and waste mishandling. During the synthesis and fabrication of such agricultural plastics, it is necessary to use chemicals, the so-called plastic additives (PAs), improving the physicochemical properties of the final polymeric product. However, since PAs are loosely bound on the polymer matrix, they can potentially leach into the soil environment with unidentified effects. Clearly, to monitor the fate of PAs in the terrestrial ecosystem, it is necessary to develop accurate, sensitive and robust analytical methods. To this end, a comprehensive analytical strategy was developed for monitoring 16 PAs with diverse physicochemical properties (partition coefficient; -3 < logP<19) in soil samples using ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). For this purpose, two different extraction procedures were developed, namely, a single step ultrasound-assisted extraction (UAE) using ethyl acetate or an aqueous solution of methanol and a binary extraction, combining Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) and UAE principles with n-hexane as the extractant. Interestingly, within the sample preparation investigation, we identified in-lab contamination sources of PAs, e.g., centrifuge tubes or microfilters. Such consumables are made of plastic contaminating the procedural blanks and omitting their use was necessary to acquire satisfactory analytical performance. In detail, method validation was performed for 16 compounds achieving recoveries mainly in the range 70-120 %, repeatability (expressed as relative standard deviation, RSD %) < 20 % and limits of quantification (LOQs) ranging between 0.2 and 20 ng/g dry weight (dw). Importantly, the presented strategies are added to the very limited available for PA determination in soil, a topical issue with a significant and rather understudied impact on agriculture.

Production and characterisation of environmentally relevant microplastic test materials derived from agricultural plastics.

Soil environments across the globe, particularly in agricultural settings, have now been shown to be contaminated with microplastics. Agricultural plastics - such as mulching films - are used in close or direct contact with soils and there is growing evidence demonstrating that they represent a potential source of microplastics. There is a demand to undertake fate and effects studies to understand the behaviour and potential long-term ecological risks of this contamination. Yet, there is a lack of test materials available for this purpose. This study describes the manufacture and characterisation of five large (1-40 kg) batches of microplastic test materials derived from agricultural mulching films. Batches were produced from either polyethylene-based conventional mulching films or starch-polybutadiene adipate terephthalate blend mulching films that are certified biodegradable in soil. Challenges encountered and overcome during the micronisation process provide valuable insights into the future of microplastic test material generation from these material types. This includes difficulties in micronising virgin polyethylene film materials. All five batches were subjected to a thorough physical and chemical characterisation - both of the original virgin films and the subsequent microplastic particles generated - including a screening for the presence of chemical additives. This is a critical step to provide essential information for interpreting particle fate or effects in scientific testing. Trade-offs between obtaining preferred particle typologies and time and cost constraints are elucidated. Several recommendations emerging from the experiences gained in this study are put forward to advance the research field towards greater harmonisation and utilisation of environmentally relevant test materials.

Assessment of organohalogenated pollutants in breast milk from the Czech Republic.

This biomonitoring survey brings new information on the occurrence of a total of 94 organohalogenated pollutants in 231 human breast milk samples collected in 2019 and 2021 from women living in two regions of the Czech Republic (Karvina and Ceske Budejovice). This study aimed to evaluate the concentrations of 6 indicator polychlorinated biphenyls (PCBs), 10 organochlorine pesticides (OCPs), 34 halogenated flame retardants (HFRs), 29 perfluoroalkyl and polyfluoroalkyl substances (PFAS) and 15 polychlorinated naphthalenes (PCNs). PCBs, OCPs, most of HFRs and PCNs were identified/quantified by gas chromatography coupled to (tandem) mass spectrometry (GC-MS(/MS)), while PFAS, hexabromocyclododecane isomers (HBCD), brominated phenols, and tetrabromobisphenol A (TBBPA) by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The mean value of the sum of the 6 indicator PCBs was 123.12 nanogram per gram of lipid weight (ng g-1 lw). Hexachlorobenzene (HCB), ß-hexachlorocyclohexane (ß-HCH) and p,p'-dichlorodiphenyl-dichloroethylene (p,p'-DDE) were the most abundant OCPs, detected in 100 % (mean 11.8 ng g-1 lw), 94.8 % (mean 6.1 ng g-1 lw) and 100 % (mean 101.5 ng g-1 lw) of samples, respectively. PCN congeners 20, 52 and 66 were detected in <1 % of the samples. The HFRs concentrations were relatively low compared to the levels of OCP; The detection rate of polybrominated diphenyl ethers (PBDEs, # 47, 99 and 153) ranged 21-68 % with a mean concentrations of 0.34 ng g-1 lw - 0.42 ng g-1 lw. PFAS concentrations were also low, with perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) dominant in this group (means of 22 pg ml-1 and 21 pg ml-1, respectively). Our results confirmed the long-term trend of declining levels of banned POPs in Czech mothers. The amounts of PCBs and OCPs were higher in older breastfeeding primiparous women.

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

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

Harmonized human biomonitoring in European children, teenagers and adults: EU-wide exposure data of 11 chemical substance groups from the HBM4EU Aligned Studies (2014-2021).

As one of the core elements of the European Human Biomonitoring Initiative (HBM4EU) a human biomonitoring (HBM) survey was conducted in 23 countries to generate EU-wide comparable HBM data. This survey has built on existing HBM capacity in Europe by aligning national or regional HBM studies, referred to as the HBM4EU Aligned Studies. The HBM4EU Aligned Studies included a total of 10,795 participants of three age groups: (i) 3,576 children aged 6-12 years, (ii) 3,117 teenagers aged 12-18 years and (iii) 4,102 young adults aged 20-39 years. The participants were recruited between 2014 and 2021 in 11-12 countries per age group, geographically distributed across Europe. Depending on the age group, internal exposure to phthalates and the substitute DINCH, halogenated and organophosphorus flame retardants, per- and polyfluoroalkyl substances (PFASs), cadmium, bisphenols, polycyclic aromatic hydrocarbons (PAHs), arsenic species, acrylamide, mycotoxins (deoxynivalenol (total DON)), benzophenones and selected pesticides was assessed by measuring substance specific biomarkers subjected to stringent quality control programs for chemical analysis. For substance groups analyzed in different age groups higher average exposure levels were observed in the youngest age group, i.e., phthalates/DINCH in children versus teenagers, acrylamide and pesticides in children versus adults, benzophenones in teenagers versus adults. Many biomarkers in teenagers and adults varied significantly according to educational attainment, with higher exposure levels of bisphenols, phthalates, benzophenones, PAHs and acrylamide in participants (from households) with lower educational attainment, while teenagers from households with higher educational attainment have higher exposure levels for PFASs and arsenic. In children, a social gradient was only observed for the non-specific pyrethroid metabolite 3-PBA and di-isodecyl phthalate (DiDP), with higher levels in children from households with higher educational attainment. Geographical variations were seen for all exposure biomarkers. For 15 biomarkers, the available health-based HBM guidance values were exceeded with highest exceedance rates for toxicologically relevant arsenic in teenagers (40%), 3-PBA in children (36%), and between 11 and 14% for total DON, Σ (PFOA + PFNA + PFHxS + PFOS), bisphenol S and cadmium. The infrastructure and harmonized approach succeeded in obtaining comparable European wide internal exposure data for a prioritized set of 11 chemical groups. These data serve as a reference for comparison at the global level, provide a baseline to compare the efficacy of the European Commission's chemical strategy for sustainability and will give leverage to national policy makers for the implementation of targeted measures.

The occurrence of perfluoroalkyl substances (PFAS) in drinking water in the Czech Republic: a pilot study.

Drinking water is one of the main contributors to overall human exposure to per- and polyfluoroalkyl substances (PFAS), a broad group of environmental contaminants with arising concerns on the impact on human health; therefore, it is necessary to monitor its quality. Here, we present a solid-phase extraction-based method to determine 22 PFAS in water, using 100 mL of the sample. The instrumental analysis employing an ultra-high-performance liquid chromatography coupled with tandem mass spectrometry achieved low limits of quantification (0.025-0.25 ng/L). The validated method (recoveries 70-120% and repeatabilities ≤ 20% at tested concentrations (0.05, 0.1 and 0.5 ng/L)) was applied to 67 tap water and 31 bottled water samples collected in the Czech Republic. The most abundant compounds were perfluorononanoic acid (88% positives; 0.034-13.3 ng/L) and perfluoroheptanoic acid (23% positives; 0.035-0.106 ng/L), respectively. ∑PFAS in positive samples ranged from 0.029 to 300 ng/L (99% positives, median 2.34 ng/L) in tap water data and 0.033 to 4.48 ng/L (32% positives, median 0.097 ng/L) in bottled water samples. Current-use fluoroalkyl ethers, dodecafluoro-3H-4,8-dioxanonanoate and 11-chloroeicosafluoro-3-oxaundecane-1-sulfonate, were occasionally detected in tap. Based on the median data, PFAS intake by an adult from a tap or bottled water represented units of % of the tolerable weekly intake set by the European Food Safety Authority and therefore did not represent a severe risk. The described method and obtained first data on PFAS in the Czech drinking water provided a solid basis for an ongoing national study on the presence of PFAS in tap water.

Critical Assessment of Clean-Up Techniques Employed in Simultaneous Analysis of Persistent Organic Pollutants and Polycyclic Aromatic Hydrocarbons in Fatty Samples.

Interference of residual lipids is a very common problem in ultratrace analysis of contaminants in fatty matrices. Therefore, quick and effective clean-up techniques applicable to multiple groups of analytes are much needed. Cartridge and dispersive solid-phase extraction (SPE and dSPE) are often used for this purpose. In this context, we evaluated the lipid clean-up efficiency and performance of four commonly used sorbents-silica, C18, Z-Sep, and EMR-lipid-for the determination of organic pollutants in fatty fish samples (10%) extracted using ethyl acetate or the QuEChERS method. Namely, 17 polychlorinated biphenyls (PCBs), 22 organochlorine pesticides (OCPs), 13 brominated flame retardants (BFRs), 19 per- and polyfluoroalkyl substances (PFAS), and 16 polycyclic aromatic hydrocarbons (PAHs) were determined in this study. The clean-up efficiency was evaluated by direct analysis in real time coupled with time-of-flight mass spectrometry (DART-HRMS). The triacylglycerols (TAGs) content in the purified extracts were significantly reduced. The EMR-lipid sorbent was the most efficient of the dSPE sorbents used for the determination of POPs and PAHs in this study. The recoveries of the POPs and PAHs obtained by the validated QuEChERS method followed by the dSPE EMR-lipid sorbent ranged between 59 and 120%, with repeatabilities ranging between 2 and 23% and LOQs ranging between 0.02 and 1.50 µg·kg-1.

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

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

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

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

Biomonitoring of 89 POPs in blood serum samples of Czech city policemen.

In this biomonitoring study, we evaluated the concentrations of 8 polychlorinated biphenyls (PCBs), 11 organochlorinated pesticides (OCPs), 33 brominated flame retardants (BFRs), 7 novel brominated and chlorinated flame retardants (novel FRs) and 30 per- and polyfluoroalkylated substances (PFAS) in human serum samples (n = 274). A total of 89 persistent organic pollutants (POPs) were measured in blood serum samples of city policemen living in three large cities and their adjacent areas (Ostrava, Prague, and Ceske Budejovice) in the Czech Republic. All samples were collected during the year 2019 in two sampling periods (spring and autumn). The identification/quantification of PCBs, OCPs, BFRs, novel FRs and PFAS was performed by means of gas chromatography coupled to (tandem) mass spectrometry (GC-MS/(MS)) and ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry (UHPLC-MS/MS). The most frequently detected pollutants were perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorooctanesulfonate (PFOS), perfluorohexanesulfonate (PFHxS), 2,2',3,4,4',5'-hexachlorobiphenyl (CB 138), 2,2',4,4',5,5'-hexachlorobiphenyl (CB 153), 2,2',3,3',4,4',5-heptachlorobiphenyl (CB 170), 2,2',3,4,4',5,5'-heptachlorobiphenyl (CB 180), hexachlorobenzene (HCB), and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) quantified in 100% of serum samples. In the serum samples, the concentrations of determined POPs were in the range of 0.108-900 ng g-1 lipid weight (lw) for PCBs, 0.106-1016 ng g-1 lw for OCPs, <0.1-618 ng g-1 lw for FRs and <0.01-18.3 ng mL-1 for PFAS, respectively. Locality, sampling season, and age were significantly associated with several POP concentrations. One of the important conclusions was that within the spring sampling period, statistically significant higher concentrations of CB 170 and CB 180 were observed in the samples from Ostrava (industrial area) compared to Prague and Ceske Budejovice. Older policemen had higher concentrations of five PCBs and two OCPs in blood serum.

The European human biomonitoring platform - Design and implementation of a laboratory quality assurance/quality control (QA/QC) programme for selected priority chemicals.

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.

Interlaboratory comparison investigations (ICI) and external quality assurance schemes (EQUAS) for cadmium in urine and blood: Results from the HBM4EU project.

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.

Comparison of polycyclic aromatic hydrocarbon metabolite concentrations in urine of mothers and their newborns.

Polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants produced during incomplete combustion of organic matter. Humans can be exposed to them via several pathways (inhalation, digestion, dermal exposure). The aim of this study was to assess the concentration of 11 monohydroxylated metabolites of PAHs (OH-PAHs) in 660 urine samples collected from mothers and their newborns residing in two localities of the Czech Republic - Most and Ceske Budejovice - in 2016 and 2017. After enzymatic hydrolysis, the target analytes were extracted from the urine samples using liquid-liquid extraction, with extraction solvent ethyl acetate and a clean-up step using dispersive solid-phase extraction (d-SPE) with the Z-Sep sorbent. For identification and quantification, ultra-high performance liquid chromatography coupled with tandem mass spectrometry was applied. 2-OH-NAP was the compound present in all of the measured samples and it was also the compound at the highest concentration in both mothers' and newborns' urine samples (median concentration 5.15 µg/g creatinine and 3.58 µg/g creatinine). The total concentrations of OH-PAHs in urine samples collected from mothers were 2 times higher compared to their children. The most contaminated samples were collected in Most in the period October 2016-March 2017 from both mothers (12.59 µg/g creatinine) and their newborns (8.29 µg/g creatinine). The concentrations of OH-PAHs in urine samples, which were collected from both mothers and their newborns as presented in this study, are comparable with those found in our previous study between 2013 and 2014. In addition, they are slightly lower or comparable to other studies from Poland, USA, Germany, China, and Australia. The results might indicate that the population in the previously highly air-polluted mining districts carries some long-term changes (maybe existing changes in genetic information), which also affect the metabolism of PAHs. It could be related to the long-lasting effect, and thus corresponding to the shortened life expectancy.