Exposure assessment of Spanish lactating mothers to acrylamide via human biomonitoring.

Acrylamide (AA) is an organic compound classified as "Probably carcinogenic to humans" (Group 2 A) that can be found principally in processed carbohydrate-rich foods and tobacco smoke. In humans, after exposure, AA is rapidly metabolized and excreted in urine, predominantly 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), which can be used as short-term biomarkers of exposure to AA. In this study, the presence of AA metabolites in urine samples of lactating mothers living in Spain (n = 114) was analyzed by "dilute and shoot" and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). All urinary metabolites were detected in 100% of the analyzed samples, with geometric means of 70, 33 and 15 ng ml-1, for AAMA, AAMA-Sul and GAMA3, respectively. The consumption of coffee, bread and precooked food products were found to be significant predictors of internal exposure to AA. An estimated daily intake (EDI) of AA based on its urinary metabolites was calculated, obtaining mean values between 1.2 and 1.9 µg AA·kg bw-1·day-1 in the target population. The risk assessment was evaluated using both reverse and forward dosimetry, showing an average margin of exposure (MOE) of 349 and a hazard quotient (HQ) of 5.5. Therefore, AA exposure should be considered a medium priority for risk assessment follow-up in the Spanish population, since a health concern with respect to non-neoplastic toxicity could not be discarded.

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).

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.

Determination of 60 Migrant Substances in Plastic Food Contact Materials by Vortex-Assisted Liquid-Liquid Extraction and GC-Q-Orbitrap HRMS.

A GC-HRMS analytical method for the determination of 60 migrant substances, including aldehydes, ketones, phthalates and other plasticizers, phenol derivatives, acrylates, and methacrylates, in plastic food contact materials (FCM) has been developed and validated. The proposed method includes migration tests, according to Commission Regulation (EU) 10/2011, using four food simulants (A, B, C, and D1), followed by vortex-assisted liquid-liquid extraction (VA-LLE) and GC-Q-Orbitrap HRMS analysis in selected ion monitoring (SIM) mode, with a resolving power of 30,000 FWHM and a mass accuracy ≤5 ppm. The method was validated, showing satisfactory linearity (R2 ≥ 0.98 from 40 to 400 µg L-1), limits of quantification (40 µg L-1), precision (RSD, 0.6-12.6%), and relative recovery (81-120%). The proposed method was applied to the analysis of field samples, including an epoxy-coated tin food can, a drinking bottle made of Tritan copolyester, a disposable glass made of polycarbonate, and a baby feeding bottle made of polypropylene, showing that they were in compliance with the current European regulation regarding the studied substances.

Biomonitoring of bisphenols A, F, S and parabens in urine of breastfeeding mothers: Exposure and risk assessment.

In the present study we used human biomonitoring to assess the internal exposure and the risk to four parabens and three bisphenols in 103 Spanish breastfeeding mothers participating in the BETTERMIILK project. Urinary methylparaben (MP), ethylparaben (EP), propylparaben (PP) and butylparaben (BP) presented detection frequencies ranging from 12% (BP) to 92% (MP), while bisphenol A (BPA), bisphenol F (BPF) and bisphenol S (BPS) were detected in 76% (BPA) and 20% (BPF, BPS) of the mothers. Average paraben concentrations (geometric mean) ranged from 0.021 ng mL-1 (BP) to 17.7 ng mL-1 (MP), whereas bisphenols had geometric means concentrations from 0.042 ng mL-1 (BPF) to 0.927 ng mL-1 (BPA). Except for BPA, the estimated daily intakes (EDI) were calculated in order to interpret urinary levels in a risk assessment context. The obtained EDIs ranged from 0.00042 mg/kg/day for PP to 0.0434 mg/kg/day for MP and EP. A hazard quotient (HQ) was calculated for BPA (0.0049) and parabens (0.001-0.004), showing no risk in the studied population. Sociodemographic characteristics, food consumption, and usage patterns of personal care products (PCPs) were investigated as possible determinants of exposure. Use of makeup and skincare products were associated with higher concentrations of MP and PP, respectively. Regarding dietary habits, MP was also associated with the consumption of packaged and bakery products.

Distributions and determinants of urinary biomarkers of organophosphate pesticide exposure in a prospective Spanish birth cohort study.

BACKGROUND: Prenatal exposure to organophosphate pesticides (OPs) has been associated with impaired child development. Pesticide exposure determinants need to be studied in order to identify sources and pathways of pesticide exposure. The aim of this paper is to describe prenatal exposure to OPs and evaluate the associated factors in pregnant women. METHODS: The study population consisted of pregnant women (n = 573) who participated in the INMA birth cohort study in Valencia (Spain, 2003-2006). OP metabolites were analyzed in maternal urine at the 32nd week of gestation using a liquid chromatography-high resolution mass spectrometry method. The analysis included non-specific (diethyl phosphate [DEP], diethyl thiophosphate [DETP], dimethyl thiophosphate [DMTP], dimethyl dithiophosphate [DMDTP]) and specific metabolites (2-diethylamino-6-methyl-4-pyrimidinol [DEAMPY], 2-isopropyl-4-methyl-6-hydroxypyrimidine [IMPY], para-nitrophenol [PNP], and 3,5,6-trichloro-2-pyridinol [TCPY]). Information about the sociodemographic, environmental, and dietary characteristics was obtained by questionnaire. The association between log-transformed OPs and covariates was analyzed using multivariable interval censored regression. RESULTS: The detection frequencies were low, DMTP and TCPY being the most frequently detected metabolites (53.8% and 39.1%, respectively). All the OP metabolites were positively associated with maternal intake of fruits and vegetables. Other maternal characteristics related to the OPs were body mass index (BMI) before pregnancy and smoking habit during pregnancy. Women with lower BMI and those who did not smoke presented higher OP concentrations. Moreover, mothers who had a yard or garden with plants at home or who lived in an urban area were also more exposed to OPs. CONCLUSIONS: The OP detection frequencies and the concentrations observed in our study population were low, compared with most of the previously published studies. Given the high vulnerability of the fetus to neurotoxicant exposure, further research on the determinants of the body burden of OPs during pregnancy would be necessary. The knowledge gained from such studies would enhance the effectiveness of public health control and future recommendations in order to reduce the risk to both the health of pregnant women and the health and development of their children.

Utilization of long duration high-volume sampling coupled to SPME-GC-MS/MS for the assessment of airborne pesticides variability in an urban area (Strasbourg, France) during agricultural application.

Atmospheric samples have been collected between 14 March and 12 September 2012 on a 2-week basis (15 days of sampling and exchange of traps each 7 days) in Strasbourg (east of France) for the analysis of 43 pesticides. Samples (particle and gas phases) were separately extracted using Accelerated Solvent Extraction (ASE) and pre-concentrated by Solid Phase Micro-Extraction (SPME) before analysis by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). Four SPME consecutive injections at distinct temperatures were made in order to increase the sensitivity of detection for the all monitored pesticides. Currently used detected pesticides can be grouped in four classes; those used in maize crops (acetochlor, benoxacor, dicamba, s-metolachlor, pendimethalin, and bromoxynil), in cereal crops (benoxacor, chlorothalonil, fenpropimorph, and propiconazole), in vineyards (tebuconazole), and as herbicides for orchards, meadows of green spaces (2,4-MCPA, trichlopyr). This is in accordance with the diversity of crops found in the Alsace region and trends observed are in accordance with the period of application of these pesticides. Variations observed permit also to demonstrate that the long time sampling duration used in this study is efficient to visualize temporal variations of airborne pesticides concentrations. Then, long time high-volume sampling could be a simple method permitting atmospheric survey of atmospheric contamination without any long analysis time and consequently low cost.

Biomonitoring exposure assessment to contemporary pesticides in a school children population of Spain.

The exposure to pesticides amongst school-aged children (6-11 years old) was assessed in this study. One hundred twenty-five volunteer children were selected from two public schools located in an agricultural and in an urban area of Valencia Region, Spain. Twenty pesticide metabolites were analyzed in children's urine as biomarkers of exposure to organophosphate (OP) insecticides, synthetic pyrethroid insecticides, and herbicides. These data were combined with a survey to evaluate the main predictors of pesticide exposure in the children's population. A total of 15 metabolites were present in the urine samples with detection frequencies (DF) ranging from 5% to 86%. The most frequently detected metabolites with DF>53%, were 3,5,6-trichloro-2-pyridinol (TCPy, metabolite of chlorpyrifos), diethyl phosphate (DEP, generic metabolite of OP insecticides), 2-isopropyl-4-methyl-6-hydroxypyrimidine (IMPY, metabolite of diazinon) and para-nitrophenol (PNP, metabolite of parathion and methyl parathion). The calculated geometric means ranged from 0.47 to 3.36 µg/g creatinine, with TCPy and IMPY showing the higher mean concentrations. Statistical significant differences were found between exposure subgroups (Mann-Whitney test, p<0.05) for TCPy, DEP, and IMPY. Children living in the agricultural area had significantly higher concentrations of DEP than those living in the urban area. In contrast, children aged 6-8 years from the urban area, showed statistically higher IMPY levels than those from agricultural area. Higher levels of TCPy were also found in children with high consumption of vegetables and higher levels of DEP in children whose parents did not have university degree studies. The multivariable regression analysis showed that age, vegetable consumption, and residential use of pesticides were predictors of exposure for TCPy, and IMPY; whereas location and vegetable consumption were factors associated with DEP concentrations. Creatinine concentrations were the most important predictors of urinary TCPy and PNP metabolites.

Levels, patterns and risk assessment of PCDD/Fs and dl-PCBs through dietary exposure in the Valencian Region (Spain).

Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin like polychlorinated biphenyls (dl-PCBs) levels were measured in representative vegetable oils and animal origin foodstuffs collected in a Total Diet Study carried out in the Valencian Region (Spain). A total amount of 3,300 food samples were collected and grouped into 5 main food groups: vegetable oils, meat and meat products, eggs, milk and dairy products and fish and sea products. The samples were analysed using gas chromatography coupled to high-resolution mass spectrometry (GC-HRMS). The food group which presented the highest concentration in wet weight (ww) for the sum of PCDD/Fs and dl-PCBs was fish and sea products (0.5 pg TEQ g-1 ww), whereas meat and meat products (0.6 pg TEQ g-1 lipid) showed the highest levels expressed in lipid terms. Occurrence data of PCDD/F and dl-PCBs were combined with consumption data to estimate the dietary exposure of adults (>15 years) and children (6-15 years). Finally, the estimated weekly intake (EWI) was calculated using a deterministic approach and considering the food consumption of the population, with fish and sea products being the main food group contributor. Likewise, considering the worst-case scenario (Upper Bound, UB), average EWI were 1.8 and 3.4 pg TEQ kg-1 body weight (bw) week-1 for adults and children, respectively. For children, the average EWI was almost twice above the tolerable weekly intake (TWI) of 2 pg TEQ kg-1 bw week-1 set by EFSA in 2018. In terms of risk characterisation, the overall obtained results showed that 19 % of adults and 43 % of children may exceed the TWI when using UB.

Indoor Air Quality and Bioaerosols in Spanish University Classrooms.

A comprehensive study assessed indoor air quality parameters, focusing on relevant air pollutants such as particulate matter (PM10 and PM2.5), gaseous compounds (CO, CO2, formaldehyde, NO2) and volatile/semi-volatile organic chemicals, as well as respiratory viruses (including SARS-CoV-2), fungi and bacteria in Spanish university classrooms. Non-target screening strategies evaluated the presence of organic pollutants inside and outside the classrooms. Saliva samples from teachers and students were collected to explore correlations between respiratory viruses in the air and biological samples. Indoor results revealed the punctual exceedance of recommended guidelines for CO2, formaldehyde (HCHO), volatile organic compounds (TVOCs) and PM in the least naturally ventilated classrooms. Significant differences occurred between the classes, with the least ventilated one showing higher average concentrations of CO2, HCHO, NO2, PM10 and PM2.5. A respiratory virus (rhinovirus/enterovirus) was detected in the medium naturally ventilated classroom, although saliva samples tested negative. Suspect screening tentatively identified 65 substances indoors and over 200 outdoors, with approximately half reporting a high toxicological risk based on the Cramer rules. The study provides a comprehensive overview of indoor air quality, respiratory viruses and organic pollutants in university classrooms, highlighting the variations and potential health risks associated with ventilation differences.

Determination of 21 per- and poly-fluoroalkyl substances in paper- and cardboard-based food contact materials by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry.

In this work, a new analytical method was developed and validated for the determination of the total content of 21 per- and poly-fluoroalkyl substances (PFASs) in paper- and cardboard-based food contact materials (FCMs). This method is based on green ultrasound-assisted lixiviation followed by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). The method was validated in various paper- and cardboard-based FCMs, showing good linearity (R2 ≥ 0.99), limits of quantification (1.7-10 µg kg-1), accuracy (74-115%), and precision (RSD < 20%). The eco-friendly characteristics of the proposed analytical method were assessed according to the analytical eco-scale, demonstrating that it can be considered an excellent green analytical method (EcoScale score > 75). Finally, 16 field samples of paper- and cardboard-based FCMs, including pizza boxes, popcorn boxes, paper bags and cardboard boxes for potato fries, ice cream tubs, pastry trays, and cardboard-based packaging for cooked Spanish omelets, fresh grapes, frozen fish, and salads were analyzed, showing that they comply with the current European regulations regarding the studied PFASs. The developed method is currently accredited according to UNE-EN ISO/IEC 17025 by the Spanish National Accreditation Body (ENAC) to be implemented for official control analysis of FCMs in the Public Health Laboratory of Valencia, Generalitat Valenciana (Valencia, Spain).

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.

Exposure and Risk Assessment to Airborne dl-PCBs and Dioxins in the Population Living in the Neighborhood of a Cement Plant: A Pilot Study in the Valencian Region of Spain.

Emissions from cement manufacturing facilities may increase health risks in nearby populations. For this reason, dioxin-like PCB (dl-PCB), polychlorinated dibenzo-p-dioxin (PCDD), and polychlorinated dibenzofuran (PCDF) concentrations in PM10 samples were assessed in the vicinity of a cement manufacturing plant located in the Valencian Region (eastern Spain). The total concentrations of the sum of dl-PCBs, PCDDs, and PCDFs ranged between 1.85 and 42.53 fg TEQ/m3 at the assessed stations. The average daily inhalation dose (DID) for the sum in adults ranged from 8.93 · 10-4 to 3.75 · 10-3 pg WHO TEQ kg-1 b.w. d-1, and, for children, the DID ranged from 2.01 · 10-3 to 8.44 · 10-3 pg WHO TEQ kg-1 b.w. d-1. Risk assessment for adults and children was performed using both daily and chronic exposure. The hazard quotient (HQ) was calculated considering 0.025 pg WHO TEQ kg-1 b.w. d-1 to be the acceptable maximum permitted inhalation exposure. The HQ obtained was slightly higher than 1 for PCDD/Fs at one of the stations (Chiva), indicating a possible health risk for the population under study due to inhalation exposure. In the case of chronic exposure, cancer risk (>10-6) was observed for some samples in one of the assessed sampling sites (Chiva).

New insights into the search of meat quality biomarkers assisted by Orbitrap Tribrid untargeted metabolite analysis and chemometrics.

Metabolite profiles of normal and defective dry, firm and dark (DFD) meat extracts with known ultimate pH (pHu) values were determined by Orbitrap Tribrid ID-X untargeted analysis coupled to chemometrics. An intelligent MS3 AcquireXTM workflow firstly approached the unambiguous characterization of detected features that were subsequently quantified by a complementary MS1 study of biological replicates. Chemometric research revealed how threonylphenylalanine (overexpressed in normal meats) together to tetradecadienoyl- and hydroxydodecanoyl-carnitines (both overexpressed in DFD meats) appropriately grouped meat groups assayed. Robustness of such biomarkers was confirmed through a time-delayed study of a blind set of samples (unknown pHu) and evidenced limitations of pHu as an isolated parameter for accurate meat quality differentiation. Other acyl-carnitines also characterized DFD samples, suggesting interferences induced by pre-slaughter stress (PSS) on lipid catabolism that would explain accumulation of such intermediate metabolites. Results achieved can ease understanding of biochemical mechanisms underlying meat quality defects.

Methods for the determination of biomarkers of exposure to emerging pollutants in human specimens.

Biomonitoring is a very useful tool for assessing human exposure to environmental pollutants. This review discusses recent methods for the quantitative measurement of biomarkers of exposure to different classes of chemicals used in personal-care products (e.g., musk fragrances, preservatives, UV filters, and insect repellents) and consumer products (e.g., organophosphate flame retardants, phthalate esters, perfluorinated compounds, and industrial phenols). The measurements are mainly taken in urine, blood, and breast milk. We also discuss the different procedures commonly used for sample-pretreatment, extraction, and clean up, and chromatographic techniques currently used to determine these compounds. Finally, we present data on the main biomarkers occurring in different human specimens.

Risk assessment of the exposure of Spanish children to acrylamide using human biomonitoring.

Acrylamide (AA) is an organic contaminant that naturally forms in starchy foods during high-temperature cooking under low-moisture conditions. It is mainly produced from the sugars and amino acids present in food by the Maillard reaction. When humans are exposed to AA, AA is eliminated in the urine as mercapturic acid conjugates, primarily including 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), which are used as exposure biomarkers of AA in human biomonitoring studies. Although the carcinogenic effects of AA on humans have not been demonstrated yet, some studies have shown that AA may negatively affect children's health. The main objective of this study was to evaluate the exposure of Spanish children (n = 612) to AA. For this purpose, the levels of AAMA, AAMA-Sul, and GAMA3 in first-morning urine samples were analyzed by "dilute and shoot" and liquid chromatography coupled to tandem mass spectrometry. The three metabolites were detected in all the children involved in this study in the following order (geometric mean (GM)): AAMA (79 ng ml-1) > AAMA-Sul (28 ng ml-1) > GAMA3 (18 ng ml-1). Statistical analysis suggested that the intake of fried potato products and biscuits could be associated with higher levels of AA metabolites in urine. Estimated daily intakes of AA in the children under study were in the range of 1.2-1.5 µg AA·kg-body weight-1·day-1 (GM). Risk assessment calculations indicate that the health risk of AA exposure cannot be overlooked and the exposure of Spanish children to AA should be closely monitored.

A green analytical method for the simultaneous determination of 30 tropane and pyrrolizidine alkaloids and their N-oxides in teas and herbs for infusions by LC-Q-Orbitrap HRMS.

A green analytical method for the simultaneous determination of 30 tropane and pyrrolizidine alkaloids and their N-oxides in dried teas and herbs for infusions has been developed and validated. The proposed method is based on QuEChERS procedure followed by LC-Q-Orbitrap HRMS analysis. The method includes a first screening analysis to assess the presence of alkaloids, followed by the quantification of suspected positive samples (cut-off level, 0.2-2.6 µg kg-1). The method was validated in five different tea and herb matrices showing satisfactory linearity (R2 ≥0.99), method limits of quantification (5 µg kg-1), accuracy (87-111 %), and precision (RSD <20 %). The greenness of the proposed method was evaluated according to the Analytical Eco-Scale, showing that it could be considered an excellent green analysis. Finally, eleven commercial field samples of tea and herbs for infusions, including rooibos, chamomile, red tea, black tea, green tea, white tea, linden, horsetail, and one infusion containing a mixture of herbs, were analyzed and the obtained results demonstrated that they were in compliance with the current European regulations regarding the studied substances.

Identification of Unknown Substances in Ambient Air (PM10), Profiles and Differences between Rural, Urban and Industrial Areas.

A fast and automated strategy has been developed for identifying unknown substances in the atmosphere (concretely, in the particulate matter, PM10) using LC-HRMS (MS3). A total of 15 samples were collected in three different areas (rural, urban and industrial). A sampling flow rate of 30 m3 h-1 was applied for 24 h, sampling a total volume of around 720 m3. A total of 49 compounds were tentatively identified using very restrictive criteria regarding exact mass, retention time, isotopic profile and both MS2 and MS3 spectra. Pesticides, pharmaceutical active compounds, drugs, plasticizers and metabolites were the most identified compounds. To verify whether the developed methodology was suitable, 11 substances were checked with their analytical standards and all of them were confirmed. Different profiles for industrial, rural and urban areas were examined. The Principal Component Analysis (PCA) model allowed us to separate the obtained data of the three assessed area. When the profiles obtained in the three evaluated areas were compared using a Volcano plot (the rural area was taken as reference), 11 compounds were confirmed as being discriminant: three of them (3-hydroxy-2-methylpyridine, 3-methyladenine and nicotine) were more likely to be found in industrial sites; ten compounds (3-hydroxy-2-methylpyridine, 3-methyladenine, azoxystrobin, cocaine, cotinine, ethoprophos, imidacloprid, metalaxyl-M, nicotine and pyrimethanil) were more probable in the case of urban sites; finally, triisopropanolamine was more likely to be detected in rural locations.

A Fast and Automated Strategy for the Identification and Risk Assessment of Unknown Substances (IAS/NIAS) in Plastic Food Contact Materials by GC-Q-Orbitrap HRMS: Recycled LDPE as a Proof-of-Concept.

A fast and automated approach has been developed for the tentative identification and risk assessment of unknown substances in plastic food contact materials (FCM) by GC-Q-Orbitrap HRMS. The proposed approach combines GC-HRMS full scan data acquisition coupled to Compound Discoverer™ 3.2 software for automated data processing and compound identification. To perform the tentative identification of the detected features, a restrictive set of identification criteria was used, including matching with the NIST Mass Spectral Library, exact mass of annotated fragments, and retention index calculation. After the tentative identification, a risk assessment of the identified substances was performed by using the threshold of toxicological concern (TTC) approach. This strategy has been applied to recycled low-density polyethylene (LDPE), which could be used as FCM, as a proof-of-concept demonstration. In the analyzed sample, 374 features were detected, of which 83 were tentatively identified after examination of the identification criteria. Most of these were additives, such as plasticizers, used in a wide variety of plastic applications, oligomers of LDPE, and substances with chemical, industrial, or cosmetic applications. The risk assessment was performed and, according to the TTC approach, the obtained results showed that there was no risk associated with the release of the identified substances. However, complementary studies related to the toxicity of the unidentified substances and the potential mixture toxicity (cocktail effects) should be conducted in parallel using bioassays.

Biomonitoring of polycyclic aromatic hydrocarbons in the urine of lactating mothers: Urinary levels, association with lifestyle factors, and risk assessment.

Polycyclic aromatic hydrocarbons (PAHs) are pollutants produced during incomplete combustion of organic matter and several industrial processes. Humans can be exposed to PAHs through ingestion of food, inhalation of tobacco smoke or polluted air, and dermal contact, causing immunologic, developmental, and reproductive problems. In the present research, eleven metabolites of PAHs were analyzed in the urine of 110 lactating women living in Spain (2015). PAH metabolites were extracted from the urine samples by liquid-liquid extraction and their determination was performed by liquid chromatography coupled to tandem mass spectrometry. In addition, information on lifestyle and dietary habits of the participants was collected using a questionnaire. All the PAH metabolites were detected in more than 70% of the samples, except for 3-hydroxybenzo(a)pyrene which was detected in less than 1% of the samples. The highest urinary levels were found for naphthalene metabolites, with geometric means of 0.8 (1-hydroxynaphthalene) and 7.1 ng ml-1 (2-hydroxynaphthalene). The statistical analysis showed that smoking status, as well as the ingestion of certain food groups (vegetables, cereals, oils and fats, smoked fish and coffee), were the main influencing factors of exposure to PAHs. The estimated daily intake (EDI) was calculated for naphthalene, fluorene, phenanthrene, and pyrene, ranging from 6 to 1522 ng kg-1·day-1. The non-cancer risk associated to PAH exposure was estimated, showing hazard quotients (HQs) and hazard indexes (HIs) below 1. Therefore, it did not reveal a significant health risk for Spanish women due to PAH exposure.