Risk assessment and predictors of the exposure to polycyclic aromatic hydrocarbons in Spanish adults by urinary human biomonitoring.

Polycyclic aromatic hydrocarbons (PAHs) are produced primarily during incomplete combustion of organic matter and in various industrial processes. They are widespread environmental pollutants that are of significant interest due to their potential toxicity. Humans can be exposed to PAHs through ingestion, inhalation and dermal contact. In the present study, ten urinary PAH biomarkers were determined in first-morning urine samples (n = 504) from the adult population (aged 18-65 years) residing in the Valencian Region of Spain. These samples were analysed using liquid-liquid extraction followed by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). All PAH biomarkers were quantified in more than 65 % of the urine samples. Naphthalene biomarkers, 1-hydroxynaphthalene (1OHNAP) and 2-hydroxynaphthalene (2OHNAP), exhibited the highest levels with geometric means (GMs) of 0.7 and 11.9 µg L-1, respectively. The 95th percentile of all PAH biomarkers ranged from 0.22 to 64.8 µg L-1. Estimated daily intakes (EDIs) for the analysed PAH families in the studied population ranged from 17 (pyrene) to 18581 (naphthalene) ng·kg-bw-1·day-1 (GM). Significant associations were observed between the quantified urinary metabolites of PAHs and smoking status, home location, annual household incomes, BMI, and the consumption of grilled food in the last 24 h. Hazard quotients (HQs) of naphthalene and consequently, hazard indexes (HIs) were above 1, especially for smokers. Therefore, potential health risks associated with PAH exposure in the target population could not be discarded.

Untargeted analysis and tentative identification of unknown substances in human tears by ultra-high performance liquid chromatography-high resolution mass spectrometry: Pilot study.

In this work, a new approach for the identification of unknown compounds in human tears has been developed and validated using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) linked to an intelligent data acquisition mode (AcquireX DS-dd-MS2) coupled to an automated data processing software (Compound Discoverer™ 3.2). As a pilot research study, four human tear samples from volunteers were analyzed. Data were acquired in both positive and negative ionization modes and exact mass, isotope pattern, and MS2 spectra match were used for the tentative identification. Following this approach, 58 substances were identified, 47 in positive mode and 11 in negative mode, with an estimated concentration ranging from 0.1 to 9000 ng mL-1. Most of them were amino acids, hormones, metabolites, and pharmaceuticals. In order to validate the proposed method, the system suitability was evaluated and 29 commercial analytical standards of the tentatively identified substances were analyzed, of which 28 were confirmed obtaining a high identification accuracy (96.6 %). These results confirm that the screening tool presented in this work can facilitate the discovery of new metabolites, novel potential biomarkers, and substances to which the person is exposed, such as pollutants.

Assessment of acrylamide exposure in Spain by human biomonitoring: Risk and predictors of exposure.

Acrylamide (AA), a chemical compound currently classified as "reasonably anticipated to be a human carcinogen", is formed through the Maillard reaction in processed carbohydrate-rich foods and is also present in tobacco smoke. The primary sources of AA exposure in the general population are dietary intake and inhalation. Within a 24-h period, humans eliminate approximately 50% of AA in the urine, predominantly in the form of mercapturic acid conjugates such as N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA), N-acetyl-S-(2-carbamoyl-2- hydroxyethyl)-L-cysteine (GAMA3), and N-acetyl-3-[(3-amino-3-oxopropyl)sulfinyl]-L-alanine (AAMA-Sul). These metabolites serve as short-term biomarkers for AA exposure in human biomonitoring studies. In this study, we analysed first-morning urine samples from the adult population (aged 18-65 years) residing in the Valencian Region, Spain, (n = 505). AAMA, GAMA-3 and AAMA-Sul were quantified in 100% of the analysed samples, with geometric means (GM) of 84, 11 and 26 µg L-1, respectively, while the estimated daily intake of AA in the studied population ranged from 1.33 to 2.13 µg·kg-bw-1·day-1 (GM). Statistical analysis of the data indicated that the most significant predictors of AA exposure were smoking and the amount of potato fried products and, biscuits and pastries consumed last 24 h. Based on risk assessment approaches conducted, the findings suggest that exposure to AA could pose a potential health risk. Therefore, it is crucial to closely monitor and continuously evaluate AA exposure to ensure the well-being of the population.

Exposure to non-persistent pesticides in the Spanish population using biomonitoring: A review.

Although Spain does not have a regular national human biomonitoring program yet, different research groups are active in evaluating the exposure of children and adults to chemicals. In the last seven years, several studies in Spain have evaluated the internal exposure of the population to currently used pesticides. The present review analyzes the scope of these studies, the employed analytical methods and the main results of the exposure and risk, mainly for children and mothers. The frequency of exposure to biomarkers of exposure to organophosphate pesticides is high. Some non-specific dialkyl phosphate metabolites (DAPs), such as the diethyl phosphate (DEP), present Detection Frequencies (DFs) in the range of 65-92% in various studies. Also, the specific biomarker of the chlorpyrifos (3,5,6-trichloro-2-pyridinol, TCPy), achieves Detection Frequencies between 74% and 100% in many studies. For pyrethroids, the metabolite 3-phenoxybenzoic acid (PBA) is present, in general, in more than the 65% of the studied samples. Highly polar herbicides were only assessed in one study and both glyphosate and its metabolite aminomethylphosphonic acid showed Detection Frequencies around 60%. However, putting the biomonitoring data in a risk assessment context, the mean Hazard Quotient (HQ), used as a metric for the individual risk, ranges from 0.0006 (glyphosate) to 0.93 in farm workers (parathion), which means that is unlike that the exposure poses a health concern (HQ < 1).

Health Risk Assessment of Exposure to 15 Essential and Toxic Elements in Spanish Women of Reproductive Age: A Case Study.

This case study investigates the exposure of 119 Spanish women of reproductive age to 5 essential (Co, Cu, Mn, V, Zn) and 10 toxic (Ba, Be, Cs, Ni, Pb, Pt, Sb, Th, Al, U) elements and assesses their risk. The essential elements (Co, Cu, Mn, V, and Zn) showed average concentrations (GM: geometric mean) of 0.8, 35, 0.5, 0.2, and 347 µg/L, respectively. Five of the toxic elements (Ba, Cs, Ni, Al, U) exhibited detection frequencies of 100%. The GM concentrations of the novel toxic elements were 12 µg/L (Al), 0.01 µg/L (Pt), 0.02 µg/L (U), 0.12 µg/L (Th), 0.009 µg/L (Be) and 4 µg/L (Cs). The urine analysis was combined with a survey to assess any variations between subgroups and potential predictors of exposure to elements in the female population. Significant differences were obtained between the rural and urban areas studied for the toxic element Cs, with higher levels found in mothers living in urban areas. In relation to diet, statistically significantly higher levels of essential (Cu) and toxic (Ba) elements were detected in women with a high consumption of fish, while mothers who consumed a large quantity of legumes presented higher levels of the toxic element Ni (p = 0.0134). In a risk-assessment context, hazard quotients (HQs) greater than 1 were only observed for the essential elements Zn and Cu in P95. No deficiency was found regarding the only essential element for which a biomonitoring equivalent for nutritional deficit is available (Zn). For the less-studied toxic elements (Al, Pt, U, Th, Be, and Cs), HQs were lower than 1, and thus, the health risk due to exposure to these elements is expected to be low for the female population under study.

Biomonitoring of Phthalates, Bisphenols and Parabens in Children: Exposure, Predictors and Risk Assessment.

Exposure to emerging contaminants, such as phthalates, bisphenols and parabens in children has been associated with possible neurodevelopment and endocrine alterations. In the present study, the biomonitoring of biomarkers in children (5-12 years old) from the Valencia Region (Spain) have been implemented using urines from the BIOVAL program. More than 75% of the children studied (n = 562) were internally exposed (>LOQ) to bisphenols and parabens, and the whole population assessed (n = 557) were exposed to at least one phthalate. The geometric means (GM) of the concentrations of bisphenol A, methyl paraben and propyl paraben were 0.9, 1.4 and 0.39 ng/mL, respectively. Regarding phthalates, monoethyl phthalate GM was 55.0 ng/mL and diethyl hexyl phthalate (as the sum of five metabolites) GM was 60.6 ng/mL. Despite the studied population being widely exposed, the detection frequencies and concentrations were in general lower than in previous studies involving children in Spain and in other countries in recent years. Furthermore, the risk assessment study concluded that the internal exposure to phthalates, bisphenols and parabens is lower than the guidance values established, and, therefore, a health risk derived from the exposure to these compounds in the studied population is not expected.

Biomonitoring of glyphosate and AMPA in the urine of Spanish lactating mothers.

The objectives of this study were to evaluate the urinary levels of Glyphosate (Gly) and its metabolite aminomethylphosphonic acid (AMPA) in Spanish breastfeeding mothers (n = 97), to identify the main predictors of exposure and to perform a risk assessment. Urine samples were analyzed using a method based on solid phase extraction and liquid chromatography tandem mass spectrometry (LC-MS/MS) determination. The developed method showed limits of quantification of 0.1 µg/L for both analytes. The detection frequencies (DFs) were 54% for Gly and 60% for AMPA, with geometric means (GMs) of 0.12 µg/L and 0.14 µg/L, respectively. In the statistical analysis, no relationship was found between the urinary levels of Gly and AMPA. Multiple regression analysis showed a significant relationship between the intake of eggs and fruits and Gly levels in urine. Estimated daily intakes (EDIs), hazard quotients (HQs), and a hazard index (HI) were calculated to analyze the obtained data from a health risk perspective. The GMs of the EDIs were 0.31 and 0.37 µg/kg of body weight (BW)/day for Gly and AMPA, respectively. The HQs were calculated considering 0.5 mg/kg BW/day as an acceptable daily intake (ADI), which EFSA has established as a health-based reference value for both analytes. The values obtained were lower than 1, and thus, low health risk due to Gly and AMPA exposure was expected for the population under study.

Liquid chromatography-Orbitrap Tribrid high-resolution mass spectrometry using data dependent-tandem mass spectrometry with triple stage fragmentation as a screening tool to perform identification and risk assessment of unknown substances in food contact epoxy resin.

A new, fast, and automatic approach has been applied for the tentative identification of unknown substances released by food contact epoxy resin after performing a migration test with food simulant. This approach combines intelligent data acquisition with AcquireX linked to liquid chromatography-Orbitrap Tribrid high-resolution mass spectrometry using data dependent-tandem mass spectrometry with triple stage fragmentation coupled to Compound Discoverer™ software for automated data processing and compound identification. The identification of the observed features was performed using a set of identification criteria, including exact mass, isotope pattern, tandem mass spectrometry spectra match, and retention time. With these criteria, 263 substances were tentatively identified. Most of the identified compounds were additives, such as plasticisers, stabilizers, and antioxidants, used in different plastic applications. However, metabolites, biological constituents with pharmacological activity, and other substances with industrial applications were also detected. In order to perform a risk assessment of the food contact epoxy resin, threshold of toxicological concern approach was applied for the identified compounds. There was not risk associated with the migration of the identified substances.

Risk assessment of exposure to phthalates in breastfeeding women using human biomonitoring.

In this study, we assessed the presence of 14 phthalate metabolites in the urine of 104 lactating mothers from Valencia (Spain) who took part in the human biomonitoring project BETTERMILK. Nine of the metabolites studied showed detection frequencies >80%, whereas the rest of the metabolites presented low detection frequencies (<5%). The concentrations ranged from

Biomonitoring of parabens in human milk and estimated daily intake for breastfed infants.

In this study, we assessed the presence of four parabens in human milk of 120 mothers from Valencia (Spain) which took part in a human biomonitoring project (BETTERMILK). The detection frequency ranges of parabens were 41-60% and 61-89% for unconjugated- and total (unconjugated + conjugated)-parabens, respectively. The concentrations ranged from

Optimization of Resolving Power, Fragmentation, and Mass Calibration in an Orbitrap Spectrometer for Analysis of 24 Pesticide Metabolites in Urine.

Mass spectrometer parameters such as Resolving Power, type of fragmentation, and mass calibration mode were optimized in the analysis of 24 pesticide metabolites in human urine using Ultra-High Pressure Liquid Chromatography coupled to Orbitrap High-Resolution Mass Spectrometer (UHPLC-HRMS). The best results were achieved with a Resolving Power of 25,000 FWHM and by applying Collision Induced Dissociation fragmentation mode (40 eV).

Biomonitoring of bisphenols A, F, S in human milk and probabilistic risk assessment for breastfed infants.

The present study addresses the presence of bisphenols A (BPA) and its analogs bisphenol F (BPF) and S (BPS) in milk of 120 mothers living in Valencia (Spain) and participating in the BETTERMILK project (year 2015). We also studied the factors that could influence the BPA levels and estimated the exposure and the risk for breast fed infants. The frequency of detection of total (conjugated + unconjugated) and unconjugated-BPA were 83% and 77%, with a geometric mean of 0.29 ng/mL and 0.15 ng/mL, respectively. The frequency of detection was much lower for total-BPF (22%) and total-BPS (1.1%). The place of residence of the mother and the use of personal care products showed significant association with BPA concentrations. The estimated daily intake of total-BPA for breastfed infants amounted to a geometric mean of 0.04 µg/kg bw and a 95th percentile of 1.0 µg/kg bw, below the tolerable daily intake of 4 µg/kg bw-day established by EFSA. To our knowledge, this is the largest biomonitoring study of bisphenols in human milk in Europe.

Determination of four parabens and bisphenols A, F and S in human breast milk using QuEChERS and liquid chromatography coupled to mass spectrometry.

For the first time, a multiresidue, sensitive and high throughput method for determination of bisphenol A, F and S and 4 parabens (methyl paraben, ethyl paraben, propyl paraben and butyl paraben) in human breast milk was developed. The proposed method includes an extraction and clean-up procedure based on QuEChERS methodology followed by liquid chromatography coupled to triple quadrupole mass spectrometry determination. Negative atmospheric pressure chemical ionization in the selected reaction monitoring mode was used for mass detection. During the method validation the recoveries varied between 83 and 115% with a precision lower than 20% for all analytes using spiked levels from 0.1 to 50 ng mL-1. The LOQ was 0.10 ng mL-1 for most of the analytes. The proposed method was successfully applied for the determination of these compounds in 10 breast milk samples from volunteer lactating mothers from the Valencian region (Spain). Among parabens, methyl paraben presented the highest detection frequency (80%) with a concentration range of 0.11-7.00 ng mL-1, while bisphenol A was detected more frequently than BPF and BPS (80% of detection frequency) with concentrations ranging from 0.13 to 1.62 ng mL-1.

Biomonitoring of mercury in hair of breastfeeding mothers living in the Valencian Region (Spain). Levels and predictors of exposure.

This study focused on the evaluation of the levels of total mercury in hair among 120 breastfeeding mothers aged 20 to 45 -. The concentrations of Hg ranged from 0.07 to 6.87 µg/g with a geometric mean (GM) of 1.22 µg/g. This GM is six times higher than the average internal exposure of mothers from other 17 European countries (0.225 µg/g). Near 70% of mothers presented levels of Hg above the USA EPA internal exposure guideline of 1 µg/g, and 27% exceeded the EFSA health-based guidance value of 1.9 µg/g. The multivariate regression analysis revealed that age, smoking and fish consumption (sword fish, small fat fish, small lean fish) were the major predictors of mercury in hair.

Retrospective analysis of pesticide metabolites in urine using liquid chromatography coupled to high-resolution mass spectrometry.

A comprehensive retrospective analysis of pesticide metabolites in urine was developed, using liquid chromatography coupled to Orbitrap high-resolution mass spectrometry (UHPLC-HRMS) that includes both post-run target (suspect screening) and non-target screening. An accurate-mass database comprising 263 pesticide metabolites was built and used for the post-run screening analysis. For non-target analysis, a "fragmentation-degradation" relationship strategy was selected. The proposed methodology was applied to 49 real urine samples from pregnant women. In the post-target analysis 26 pesticide metabolites were tentatively identified, 8 of which (2-diethylamino-6-methyl-pyrimidinol; 3-ketocarbofuran; 4,6-dimethoxy-2-pyrimidinamine; 4-hydroxy-2-isporopyl-6-methylpyrimidine; diethyl malate; diethyl maleate; N-(2-Ethyl-6-methylphenyl)-2-hydroxyacetamide and propachlor oxanilic acid ) were confirmed using analytical standards. Likewise, one unknown degradation product, methyl-N-phenylcarbamate was elucidated in the non-target screening. Finally, the real urine samples were grouped according to their origin applying a metabolomic approach.