Association among serum per- and polyfluoroalkyl substances, lipid profile and metabolic syndrome in Czech adults, HBM-EHES survey 2019.

OBJECTIVES: Per- and polyfluoroalkyl substances (PFASs) are a large group of persistent synthetic chemicals widely used commercially. They accumulate increasingly in all environmental components and enter the organisms, including humans. Some of them are associated with the risk of harm to health, among others with metabolic disorders. To test the associations between blood serum levels of PFASs and blood lipid profile as well as metabolic syndrome, we linked human biomonitoring with the Czech Health Examination Survey (CZ-EHES) conducted in 2019. METHODS: A total of 168 participants of the CZ-EHES survey aged 25-64 years were examined including anthropometrical data and analyses for serum PFAS and blood lipid levels. Extended model approach in multiple linear regression models was used for identification of the associations between serum levels of 11 PFASs and lipid profile components. The relation between PFAS serum levels and metabolic syndrome prevalence was tested using a logistic regression model. RESULTS: Six PFASs were detected over the limit of quantification in at least 40% cases and were examined in subsequent analyses: perfluorodecanoic acid (PFDA), perfluorohexane sulfonic acid (PFHxS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorooctane sulfonic acid (PFOS), perfluoroundecanoic acid (PFUdA). The most dominant was PFOS with the mean value amounting to 4.81 ng/ml. After adjusting for potential confounders, we found a significant positive association between serum PFHxS and blood total cholesterol (p = 0.005) as well as LDL-cholesterol (p = 0.008). Significant positive association was also found between PFDA and HDL-cholesterol levels (p = 0.010). No significant associations were detected between PFASs and triglycerides, and between PFASs and metabolic syndrome. CONCLUSIONS: We found some evidence of a significant association between blood serum PFAS levels and blood cholesterol levels. Our results did not confirm an association between serum PFASs and the metabolic syndrome prevalence.

Exposure to Mixed Phthalates in Czech Preschool and School Children.

Knowledge of population exposure to phthalates based on the urinary metabolite levels is of the highest importance for health risk assessment. Such data are scarce in the Czech population. In the study conducted in 2016, six urinary phthalate metabolites were analysed in a total of 370 first morning urine samples of healthy children aged 5 and 9 years, namely mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (5OH-MEHP), mono(2-ethyl-5-oxohexyl) phthalate (5oxo-MEHP), mono-benzyl phthalate (MBzP), mono-iso-butyl phthalate (MiBP), and mono-n-butyl phthalate (MnBP). The two latter mono-butyl phthalate isoforms dominated among all samples with geometric means of 63.0 µg/L (MnBP) and 44.1 µg/L (MiBP), followed by 5OH-MEHP (20.6 µg/L), 5oxo-MEHP (12.9 µg/L), MBzP (3.65 µg/L), and MEHP (2.31 µg/L). Daily intake (DI) of the parent phthalates was estimated using the creatinine-based model. The highest DI values were found for di-n-butyl phthalate (DnBP) (median 2.5 µg/kg bw/day; 95th percentile 7.8 µg/kg bw/day) and di-2-ethylhexyl phthalate (DEHP) (median 2.3 µg/kg bw/day; 95th percentile 8.9 µg/kg bw/day) in 5-year-old children. The tolerable daily intake (TDI) set by the European Food Safety Authority (EFSA) was exceeded in case of DnBP (in 1% of 9-year-olds and in 3% of 5-year-olds). Exposure risk was assessed based on hazard quotients calculation and cumulative approach for similar health effect. The combined exposure to four phthalates expressed by hazard index (HI) for reprotoxicity revealed exceeding of HI threshold in 14% of 5-year-olds and in 9% of 9-year-olds. These findings strongly support the need to reduce the burden of children by phthalates.

Alternative methods in vitro for screening of endocrine disruptors.

OBJECTIVES: The aim of this study was to compare in silico data with results obtained in two alternative in vitro methods; and to investigate the potential endocrine activity of bisphenol A analogues. This article contributes to recent findings and brings up-to-date information on development of EU legislation and in vitro testing methods of endocrine disruption. METHODS: In silico approach based on the OECD QSAR Toolbox was used for prediction of potential ligands of human estrogen receptor α. Estrogen Receptor Transactivation in vitro Assay to Detect Estrogen Receptor Agonists and Antagonists (OECD TG 455/457) using the VM7Luc4E2 (formerly designated BG1Luc4E2) cell line was performed for measurement of transactivation activity of the tested substances. Commercially available yeast-based microplate assay (XenoScreen YES/YAS, Xenometrix, Switzerland) for detection of compounds with estrogenic and androgenic agonistic/antagonistic activity was used as a comparative test to estrogen receptor transactivation assay (OECD TG 455/457) and for screening of the agonistic/antagonistic potential of human estrogen receptor and agonistic/antagonistic activity of tested compounds on human androgen receptor. RESULTS: The study showed good correlation between the two in vitro assays and significant correlation with in silico data. All tested substances were identified as agonists for human estrogen receptor α by methods in silico and in vitro, four substances showed a potentially higher estrogenic activity comparing to bisphenol A, two substances were identified as very weak antagonists of human androgen receptor and one compound showed a potential of agonistic activity to human androgen receptor. CONCLUSIONS: The study contributes to recent findings and brings new in silico and in vitro data of bisphenol A analogues, revealing that these analogous substances should be further tested as they may show similar or higher activity in vivo comparing to bisphenol A, which has been recently legislatively regulated.

Urine Levels of Phthalate Metabolites and Bisphenol A in Relation to Main Metabolic Syndrome Components: Dyslipidemia, Hypertension and Type 2 Diabetes. A Pilot Study.

AIM: Human exposure to organic pollutants (some of them also called endocrine disruptors) can be associated with adverse metabolic health outcomes including type 2 diabetes. The goal of this study was to compare the urine levels of bisphenol A and phthalate metabolites in subgroups of patients with metabolic syndrome composed of patients with and without three important components of metabolic syndrome (hypertension, dyslipidemia and diabetes). METHODS: We have investigated 24 hr urine samples of 168 patients with metabolic syndrome from the Metabolic Outpatient Department of General University Hospital in Prague. Using standard metabolic syndrome criteria, we classified patients as dyslipidemic (n=87), hypertensive (n=96), and type 2 diabetic (n=58). Bisphenol A and 15 metabolites of phthalates were evaluated in relation to creatinine excretion. Samples were analysed with enzymatic cleavage of glucuronide using ultra-high-performance liquid chromatography-electrospray ionization tandem mass spectrometry in one laboratory with external quality control. RESULTS: Four metabolites, mono-n-butyl phthalate, mono-(2-ethyl-5-hydroxyhexyl) phthalate, mono-(2-ethyl-5-oxohexyl) phthalate, and mono-(2-ethyl-5-carboxypentyl) phthalate showed significantly higher levels in diabetic compared to non-diabetic patients (p<0.001, p=0.002, p=0.002, and p=0.005, respectively). The differences remained significant after adjustment to hypertension, dyslipidemia, age, and BMI. No difference was found between either the hypertensive and non-hypertensive or dyslipidemic and non-dyslipidemic patients. There was no significant relation of bisphenol A level to diabetes, hypertension, dyslipidemia, age, and BMI. CONCLUSIONS: Urine levels of four phthalate metabolites were significantly higher in type 2 diabetics independently on specified predictors. Phthalate levels can be in relation to beta cell dysfunction in type 2 diabetic patients but this study is not able to show if the relation is causal.

[Potential sources of phthalates and bisphenol A and their significance in the development of metabolic diseases]. / Potenciální zdroje ftalátu a bisfenolu A a jejich význam u metabolických onemocnení.

Nowadays, there is increasing evidence showing that the development of the metabolic syndrome combining obesity, type 2 diabetes mellitus, arterial hypertension and dyslipidemia involves except of traditional risk factors (overnutrition, lack of physical activity, genetic predisposition) also the effect of environmental organic substances called organic pollutants or endocrine disruptors. These chemicals can be found in plastic covers, paints, flame retardants, exhaust gases, fertilizers as well as diverse daily utensils. Phthalates, used primarily as plasticizers, and bisphenol A, are among the most wide-spread members of this group.The aim of this article is to provide a basic overview of the relationship between phthalates and bisphenol A and the etiopathogenesis of the metabolic syndrome and to highlight their potential sources. According to the analysis of materials used for parenteral nutrition and urinary excretion of phthalate metabolites and bisphenol A in subjects on long-term parenteral nutrition we suppose that currently used medical materials are safe with respect to the exposure to both phthalates and bisphenol A and that home environment, especially cosmetic products, might constitute a more probable source of these substances.

New approach methods for assessing indoor air toxicity.

Indoor air is typically a mixture of many chemicals at low concentrations without any adverse health effects alone, but in mixtures they may cause toxicity and risks to human health. The aim of this study was by using new approach methods to assess the potential toxicity of indoor air condensates. In specific, different in vitro test methods including cyto-and immunotoxicity, skin sensitization and endocrine disruption were applied. In addition to biological effects, the indoor air samples were subjected to targeted analysis of 25 volatile organic compounds (VOCs) and Genapol X-80 (a nonionic emulsifier) suspected to be present in the samples, and to a non-targeted "total chemical scan" to find out whether the chemical composition of the samples is associated with the biological effects. The results confirm that assessing health risks of indoor air by analysing individual chemicals is not an adequate approach: We were not able to detect the VOCs and Genapol X-80 in the indoor air samples, yet, several types of toxicity, namely, cytotoxicity, immunotoxicity, skin sensitization and endocrine disruption were detected. In the non-targeted total chemical scan of the indoor air samples, a larger number of compounds were found in the cytotoxic samples than in the non-cytotoxic samples supporting the biological findings. If only one biological method would be selected for the screening of indoor air quality, THP-1 macrophage/WST-1 assay would best fit for the purpose as it is sensitive and serves as a good representative for different sub-toxic end points, including immunotoxicity, (skin) sensitization and endocrine disruption.

Monitoring of pesticides in drinking water: finding the right balance between under- and over-monitoring - experience from the Czech Republic.

The modern, risk-based approach requires that only those pollutants which are likely to be present in a given water supply should be monitored in drinking water. From this perspective, defining an adequate approach to the monitoring of pesticides in areas with intensive agriculture is currently one of the greatest issues of regulation. This article shows the development and detailed results of pesticide monitoring in drinking water in the Czech Republic (CR). More than 4000 water supply zones serving around a 9.5 million population are routinely monitored, with nearly 250 thousand analyses of over 200 different pesticides and their metabolites being performed every year, with a non-compliance rate of ca. 0.3%. In 2017, pesticides accounted for most derogations in the CR, concerning a total of 64 water supply systems serving more than a 250 thousand population. A representative survey targeting 21 selected chemicals showed that 75% of water supply systems contained up to 11 pesticides per sample. The most commonly found were metabolites of the herbicides used to protect oilseed rape, maize, and sugar beet: acetochlor ESA, alachlor ESA, metazachlor OA, and chloridazon-desphenyl. The health risk assessment did not reveal any risks from these chemicals, even at the highest levels detected or in the most abundant mixtures, to the most vulnerable population (infants). Nevertheless, the increased presence of pesticides undermines the public's trust in drinking water safety.

Easy and Inexpensive Method for Multiclass Analysis of 41 Food Contact Related Contaminants in Fatty Food by Liquid Chromatography-Tandem Mass Spectrometry.

Food contact materials (FCMs) may release their chemical components into food and thus raise safety concerns. This paper attempted to study the presence of four major groups of FCM-related endocrine disruptors in fatty food: dialkyl phthalates, bisphenols, printing ink photoinitiators, and polyfluoroalkyl substances. All 41 target compounds were analyzed simultaneously by means of liquid chromatography coupled to tandem mass spectrometry. The sample preparation was significantly streamlined to reduce analysis costs by employing acetonitrile extraction, extract modification by water, and refrigeration at 5 °C. The new method was validated and applied to 60 real samples, including edible oils, butter, and chocolate, where 16 target compounds were measured at levels ≤13000 ng/g. The study also described the blank level increase and sensitivity loss caused by impurities present in the HPLC methanol solvent.

Toxicity of food contact paper evaluated by combined biological and chemical methods.

The study was focused on assessment of potential health risks of paper-based food contact materials (FCMs) in a step-wise approach using three toxicological bioassays in vitro and chemical analyses of migrating contaminants. 3T3 NRU cytotoxicity test showed high sensitivity to detect basal toxicity of FCMs extracts and served as a first-line test for selection of samples for further testing. The reconstructed human intestine model EpiIntestinal showed more realistic tissue response than cell culture monolayer and higher resistance despite prolonged exposure to the selected 6 samples, i.e. negligible decrease of viability and intestinal penetration, nevertheless an increase of IL-8 after exposure to black printed sample extract. Yeast based assays identified weak agonistic/antagonostic activity to human androgen receptor of the black printed sample. In accordance with the biological effects, the targeted LC and GC analytical methods confirmed the presence of high amounts of phthalates, photoinitiators and PAHs that could justify the hazard of the black printed sample. Heavily printed uncoated FCMs are recognized not to be suitable for direct contact with food. The selected bioassays and chemical analyses might be useful tools to detect targeted biological effects of xenobiotics suspected to contribute to human exposure from food.

Solution for blank and matrix difficulties encountered during phthalate analysis of edible oils by high performance liquid chromatography coupled with tandem mass spectrometry.

Worldwide production of phthalates has led to their undesirable presence in the food chain. Particularly edible oils have become an area of growing concern owing to numerous reported occurrences of phthalates. The analytical methods used in this field face difficulties associated mainly with matrix complexity or phthalate contamination which this study has aimed to describe and resolve. The proposed procedure consisting of liquid-liquid extraction, solid phase extraction and high performance liquid chromatography coupled with tandem mass spectrometry allowed us to analyze simultaneously 6 individual phthalates and 2 phthalate isomeric mixtures. DSC-18 SPE phase was selected for cleanup owing to the most efficient co-extract removal (assessed using high resolution mass spectrometry). Several sources of phthalate contamination were identified, however, the mobile phase was the most serious. The key improvement was achieved by equipping a contamination trap, a 50-mm reverse phase HPLC column, generating a delay between target and mobile phase peaks of the same compounds. RSDs ranging between 2.4 and 16 % confirm good precision and LOQs between 5.5 and 110µgkg(-1) reflect satisfactory blank management. With up to 19 occurrences in 25 analyzed edible oil samples and levels up to 33mgkg(-1), bis(2-ethylhexyl), diisononyl and diisodecyl phthalates were the most important contaminants.