Assessment of genotoxic damage induced by exposure to binary mixtures of polycyclic aromatic hydrocarbons and three heavy metals in male mice.

INTRODUCTION: Heavy metals (HM) and polycyclic aromatic hydrocarbons (PAHs) exposition has been associated with health problems. Therefore, this research evaluated genotoxicity induced in male mice strain CD-1 exposed to benzo[a]anthracene (B[a]A) and benzo[a]pyrene (B[a]P) and their interaction with Fe, Pb, and Al. METHODS: Groups of animals were exposed intraperitoneally to HM, PAHs, and mixtures of both. Peripheral blood samples were taken from 0 to 96 h at 24 h intervals; genotoxicity was determined by micronucleus tests and comet assay. Additionally, toxicity and viability were evaluated. RESULTS: HM and PAHs individually were genotoxic. About toxicity, only Al altered polychromatic erythrocytes number and did not change leukocytes viability. Concerning mixtures, Fe + B[a]P, Fe + B[a]A, Pb + B[a]P increased genotoxicity. There were no changes with Pb + B[a]A. Finally, Al mixtures with both PAHs damage was decreased. CONCLUSIONS: Exposure to HM and PAH caused genetic damage. Fe, Al, and B[a]A, established a genotoxic potential. Every metal can interact with PAHs in different ways. Also, the micronucleus test and the comet assay demonstrated their high capacity and reliability to determine the genotoxic potential of the compounds evaluated in this work.

Quantitative immunochromatographic assay for rapid and cost-effective on-site detection of benzo[a]pyrene in oilfield chemicals.

Contamination of oilfield chemicals (OFCs) by benzo[a]pyrene (B[a]P) is increasingly becoming a severe environmental security issue. There is an urgent need to develop a rapid and accurate method for B[a]P detection in OFCs. In this study, B[a]P hapten was designed using computer aided molecular design. A high-affinity, specific, and matrix-insensitive monoclonal antibody (mAb) with IC50 values of 6.77 ng/mL was obtained. Based on this mAb, we developed a rapid gold nanoparticle-based immunochromatographic strip assay (GICA) with double T-line mode for on-site detection of B[a]P in OFCs samples. The GICA exhibited excellent detection performance in OFCs samples with strong acidity, strong alkalinity, and deep color. Under optimal conditions, the proposed method detected B[a]P in OFCs at 0.42-300 mg/kg, and limit of detection was 0.23-1.07 mg/kg. The recovery rate was 88-106% with a coefficient of variation of 1.46-6.35%. Confirmed by natural positive OFCs samples and high-performance liquid chromatography, this GICA is accurate and reliable, with great potential for rapid and cost-effective on-site detection.

New insight into PAH4 induced hepatotoxicity and the dose-response assessment in rats model.

PAH4 (sum of benzo[a]pyrene, chrysene, benz[a]anthracene and benzo[b]fluoranthene) has been proposed as better marker than benzo[a]pyrene to assess total PAHs exposure in foodstuffs. However, the toxicological behaviors of PAH4 combined exposure remain unclear. This study aimed to investigate PAH4 toxicity effects with non-targeted metabolomics approach and evaluate the external and internal dose-response relationships based on benchmark dose (BMD) analysis. Male Sprague-Dawley rats were treated by gavage with vehicle (corn oil) or four doses of PAH4 (10, 50, 250, 1000 µg/kg·bw) for consecutive 30 days. After the final dose, the liver, blood and urine samples of rats were subsequently collected for testing. The concentrations of urinary mono-hydroxylated PAHs metabolites (OH-PAHs) including 3-hydroxybenzo[a]pyrene (3-OHB[a]P), 3-hydroxychrysene (3-OHCHR) and 3-hydroxybenz[a]anthracene (3-OHB[a]A) were determined to reflect internal PAH4 exposure. Our results showed PAH4 exposure increased relative liver weight and serum aspartate aminotransferase (AST) activity and caused hepatocyte swelling and degeneration, implying hepatotoxicity induced by PAH4. Serum metabolomics suggested PAH4 exposure perturbed lipid metabolism through upregulating the expression of glycerolipids metabolites, which was evidenced by markedly increased serum triglyceride (TG) level and hepatic TG content. Additionally, urinary OH-PAHs concentrations presented strong positive correlations with the external dose, indicating they were able to reflect PAH4 exposure. Furthermore, PAH4 exposure led to a dose-response increase of hepatic TG content, based on which the 95% lower confidence value of BMDs for external and internal doses were estimated as 5.45 µg/kg·bw and 0.11 µmol/mol·Cr, respectively. In conclusion, this study suggested PAH4 exposure could induce hepatotoxicity and lipid metabolism disorder, evaluating the involved dose-response relationships and providing a basis for the risk assessment of PAHs.

Polycyclic aromatic hydrocarbons in the Chinese diet: contamination characteristics, indicator screening, and health risk assessment.

Polycyclic aromatics hydrocarbons (PAHs) are ubiquitous in foods and environment and possess carcinogenic and mutagenic potential. Foods are the main source of exposure to PAHs in the general population. In this study, we determined the concentrations of 16 European Union priority PAHs in 1,564 foodstuffs acquired from nine provinces and commonly consumed by the Chinese population. The most predominant PAH was chrysene (16.7%), followed by benz[a]anthracene (12.4%) and benzo[b]fluoranthene (11.7%). Edible vegetable oils (17.89 µg/kg) and fruits (1.97 µg/kg) had the highest and lowest concentrations of total PAHs, respectively. Suitable indicators of PAH contamination in foods were assessed based on the occurrence of other PAHs in samples negative for benzo[a]pyrene and the correlation for the PAHs and their combinations. According to our results, PAH4 was a suitable indicator, better than PAH8 and benzo[a]pyrene alone. PAH exposure in the Chinese population was estimated by combining contamination data with national individual food consumption data, based on the middle bound approach. The overall average dietary exposures for BaP and PAH4 were 3.08 and 17.61 ng/kg bw/day, respectively. The major contributors to the total dietary exposure of PAHs were cereals (39%), edible vegetable oils (28%), and vegetables (20%). We used the margin of exposure (MOE) approach to assess health risk for consumers. MOEs of the mean estimated dietary exposures were >10,000, indicating a low concern for the health of the general population and of consumers of smoked, grilled, or barbecued foods. For high consumers and children, the MOEs were <10,000, indicating potential concerns.

Modelling benzo(a)pyrene concentrations for different meteorological conditions - Analysis of lung cancer cases and associated economic costs.

Air pollution originating from the household presents a significant burden to public health, especially during the wintertime in countries, such as Poland, where coal substantially contributes to the energy market. One of the most hazardous components of particulate matter is benzo(a)pyrene (BaP). This study focusses on the impact of different meteorological conditions on BaP concentrations in Poland and associated impacts on human health and economic burdens. For this study, we used the EMEP MSC-W atmospheric chemistry transport model with meteorological data from the Weather Research and Forecasting model to analyze the spatial and temporal distribution of BaP over Central Europe. The model setup has two nested domains, with the inner domain at 4 km × 4 km over Poland, which is a hotspot for BaP concentrations. The outer domain covers countries surrounding Poland in coarser resolution (12 × 812 km), to ensure that transboundary pollution is properly characterized in the modelling. We investigated the sensitivity to variability in winter meteorological conditions on BaP levels and impacts using data from 3 years: 1) 2018, which represents average meteorological conditions during the winter season (BASE run), 2) 2010 with a cold winter (COLD), and 3) 2020 with a warm winter (WARM). The ALPHA-RiskPoll model was used to analyze the lung cancer cases and associated economic costs. The results show that the majority of Poland exceeds the target level of benzo(a)pyrene (1 ng m-3) mainly due to high concentrations during the cold months. High concentrations of BaP have serious health implications and the number of lung cancers in Poland due to BaP exposure varies from 57 to 77 cases for the WARM and COLD years, respectively. It is reflected in the economic costs, which ranged from 136, through 174 to 185 million euros/year for the WARM, BASE and COLD model runs, respectively.

A multi-pathway exposure assessment for polycyclic aromatic hydrocarbons among residents in the Athabasca oil sands region, Canada.

Due to increasing emissions from ongoing development of the oil sands in Northern Alberta, Canada, there is concern that local residents and organisms are experiencing elevated exposures to hazardous contaminants. We modified an existing human bioaccumulation model (ACC-Human) to represent the local food chain in the Athabasca oil sands region (AOSR), the focus of oil sands development in Alberta. We used the model to assess the potential exposure to three polycyclic aromatic hydrocarbons (PAHs) among local residents that have a high intake of locally sourced traditional foods. To place these estimates into context, we complemented them with estimated PAH intake through market foods and smoking. Our approach was able to produce realistic body burdens of the PAHs in aquatic and terrestrial wildlife and in humans, both in magnitude and with respect to the relative difference between smokers and non-smokers. Over the model simulation period (1967-2009), market food was the dominant dietary exposure route for phenanthrene and pyrene, while local food, and in particular local fish, dominated the intake of benzo[a]pyrene. Exposure to benzo[a]pyrene therefore was also predicted to increase over time in concert with expanding oil sands operations. Those smoking at the average rate of Northern Albertans take in an additional amount of all three PAHs that is at least as large as dietary intake. Estimated daily intake rates are below toxicological reference thresholds for all three PAHs. However, daily intake of BaP in adults is only ∼20 fold below those thresholds and is predicted to increase. Key uncertainties in the assessment included the effect of food preparation on the PAH content in food (e.g., smoking of fish), the limited availability of market food contamination data specific to Canada, and the PAH content of the vapor phase of first-hand cigarette smoke. Considering the satisfactory model evaluation, ACC-Human AOSR should be suited to making predictions of future contaminant exposure based on development scenarios in the AOSR or in response to potential emission reduction efforts. It should also be applicable to other organic contaminants of concern released by oil sands operations.

Characterization, risk assessment, and source estimation of PM10-bound polycyclic aromatic hydrocarbons during wintertime in the ambient air of Basrah City, Iraq.

In this study, the concentration and structure of polycyclic aromatic hydrocarbons (PAHs) associated with the ambient PM10 in Basrah City, Iraq have been investigated for the first time. From December 2021 to February 2022, PM10 samples were collected on quartz fiber filters, extracted using an optimized extraction protocol, and analyzed for the sixteen US EPA priority PAHs. The results indicated that 4- and 5-ring PAHs represent 52% of the total detected PAHs. The most abundant PAHs over the study period were chrysene (1.2 ± 1.5 ng m-3), fluorene (0.9 ± 1.4 ng m-3), and benzo[b]fluoranthene (0.7 ± 0.9 ng m-3). Source identification suggested that PM10-bound PAHs primarily originated from pyrogenic and petrogenic activities in Basrah City. In addition, the cancer risk associated to PAH exposure was assessed based on benzo[a]pyrene equivalent concentration and was found ranging from 0.07 to 6.32 ng m-3; hence, it exceeded the threshold limit of 1.0 ng m-3 established by the European legislation (EU, 2014). Benzo[a]pyrene was determined to be main contributor to total carcinogenic power of the detected PAHs, accounting for 50.3%, followed by dibenz[a,h]anthracene (22.3%). Similarly, benzo[a]pyrene represented a major contributor to PAH associated mutagenicity, accounting for 43.5% of the total.

Identification and Risk Assessment of Priority Control Organic Pollutants in Groundwater in the Junggar Basin in Xinjiang, P.R. China.

The Junggar Basin in Xinjiang is located in the hinterland of Eurasia, where the groundwater is a significant resource and has important ecological functions. The introduction of harmful organic pollutants into groundwater from increasing human activities and rapid socioeconomic development may lead to groundwater pollution at various levels. Therefore, to develop an effective regulatory framework, establishing a list of priority control organic pollutants (PCOPs) is in urgent need. In this study, a method of ranking the priority of pollutants based on their prevalence (Pv), occurrence (O) and persistent bioaccumulative toxicity (PBT) has been developed. PvOPBT in the environment was applied in the screening of PCOPs among 34 organic pollutants and the risk assessment of screened PCOPs in groundwater in the Junggar Basin. The results show that the PCOPs in groundwater were benzo[a]pyrene, 1,2-dichloroethane, trichloromethane and DDT. Among the pollutants, benzo[a]pyrene, 1,2-dichloroethane and DDT showed high potential ecological risk, whilst trichloromethane represented low potential ecological risk. With the exception of benzo[a]pyrene, which had high potential health risks, the other screened PCOPs had low potential health risks. Unlike the scatter distribution of groundwater benzo[a]pyrene, the 1,2-dichloroethane and trichloromethane in groundwater were mainly concentrated in the central part of the southern margin and the northern margin of the Junggar Basin, while the DDT in groundwater was only distributed in Jinghe County (in the southwest) and Beitun City (in the north). Industrial and agricultural activities were the main controlling factors that affected the distribution of PCOPs.

Replacing animal-derived components in in vitro test guidelines OECD 455 and 487.

The evaluation of single substances or environmental samples for their genotoxic or estrogenic potential is highly relevant for human- and environment-related risk assessment. To examine the effects on a mechanism-specific level, standardized cell-based in vitro methods are widely applied. However, these methods include animal-derived components like fetal bovine serum (FBS) or rat-derived liver homogenate fractions (S9-mixes), which are a source of variability, reduced assay reproducibility and ethical concerns. In our study, we evaluated the adaptation of the cell-based in vitro OECD test guidelines TG 487 (assessment of genotoxicity) and TG 455 (detection of estrogenic activity) to an animal-component-free methodology. Firstly, the human cell lines A549 (for OECD TG 487), ERα-CALUX® and GeneBLAzer™ ERα-UAS-bla GripTite™ (for OECD TG 455) were investigated for growth in a chemically defined medium without the addition of FBS. Secondly, the biotechnological S9-mix ewoS9R was implemented in comparison to the induced rat liver S9 to simulate in vivo metabolism capacities in both OECD test guidelines. As a model compound, Benzo[a]pyrene was used due to its increased genotoxicity and endocrine activity after metabolization. The metabolization of Benzo[a]Pyrene by S9-mixes was examined via chemical analysis. All cell lines (A549, ERα-CALUX® and GeneBLAzer™ Erα-UAS-bla GripTite™) were successfully cultivated in chemically defined media without FBS. The micronucleus assay could not be conducted in chemically defined medium due to formation of cell clusters. The methods for endocrine activity assessment could be conducted in chemically defined media or reduced FBS content, but with decreased assay sensitivity. The biotechnological ewoS9R showed potential to replace rat liver S9 in the micronucleus in FBS-medium with A549 cells and in the ERα-CALUX® assay in FBS- and chemically defined medium. Our study showed promising steps towards an animal-component free toxicity testing. After further improvements, the new methodology could lead to more reproducible and reliable results for risk assessment.

Health Risk Assessment of Dermal Exposure to Polycyclic Aromatic Hydrocarbons from the Use of Infant Diapers.

In September 2021, the European Chemicals Agency evaluated a dossier for restricting polycyclic aromatic hydrocarbons (PAHs) in infant diapers and concluded that risks were not demonstrated, because of inconclusive exposure data. To fill this gap, we measured the 16 priority PAHs of the U.S. Environmental Protection Agency in the diaper core of four brands and in the sheets and fastening tapes of six brands of commercially available diapers. Health risks were conservatively assessed by assuming that dermally absorbed PAHs can cause both local (skin cancer) and systemic critical effects (neurobehavioral changes). Total concentrations of PAHs in the diaper core and top sheet, the only significant contributors to skin exposure, averaged 26.5 µg/kg and 66.6 µg/kg, respectively. Excess skin cancer risks and hazard quotients for neurobehavioral effects calculated with the daily dose of total PAHs from the combined diaper core and top sheet averaged 1.44 × 10-7 and 1.19 × 10-2, respectively. The median daily doses of total PAHs and of its benzo[a]pyrene-equivalent from breast milk estimated worldwide are 171 and 30 times greater than that from the combined diaper core and top sheet, respectively. Altogether, these findings indicate that trace levels of PAHs found in infant diapers are unlikely to pose health risks.

Deriving freshwater sediment quality guidelines of polycyclic aromatic hydrocarbons using method of species sensitivity distribution and application for risk assessment.

Contamination of polycyclic aromatic hydrocarbons (PAHs) in sediment has long been of great concern because of their toxic effects to benthic organisms. Sediment quality guidelines (SQGs) are the basis to evaluate the potential ecological risks of PAHs in sediments. Species sensitivity distribution (SSD) has been widely applied in deriving water quality criteria, but seldom employed in SQGs. In this study, SSD was used to derive the freshwater SQGs for four representative PAHs (naphthalene, phenanthrene, pyrene and benzo[a]pyrene) based on the sediment toxicity results. A linear relationship between the SQGs and octanol-water partition coefficient (log KOW) was developed, and applied to predict the SQGs of other twelve PAHs. The obtained SQGs were in the range of 0.46 - 1.79 mg/kg with a geometric mean of 0.97 mg/kg, which was proposed as the SQGs for total PAHs. Based on these SQGs, the risk quotients of PAHs in the sediments collected from Haihe River of China were calculated, and the toxic effects were also tested using three representative benthic organisms. As the risk quotients of the PAHs and heavy metals in the sediments were summed up, good correlations were found (p = 0.074 and 0.018) between them and the observed toxicities of the sediments. The SQGs developed for PAHs was promising in ecological risk assessment for contaminated freshwater sediments.

Assessment of the exposure to polycyclic aromatic hydrocarbons in users of various tobacco/nicotine products by suitable urinary biomarkers.

Polycyclic aromatic hydrocarbons (PAHs) occur naturally (bitumen and oils) and are formed during all incomplete combustions of organic materials. PAH exposure sources are manifold and include specific workplaces, ambient air, various foodstuffs, tobacco smoke and some medications. At least four members of this class of chemicals have been classified as proven or probable human carcinogens. Assessment of the exposure to PAHs with suitable methods is of importance, particularly in users of new-generation tobacco/nicotine products, which are intended to replace combustible cigarettes (CCs), a major source of non-occupational exposure to PAHs. In a clinical study comprising a period of 74 h under confinement, we investigated the exposure to naphthalene (Nap), fluorene (Flu), phenanthrene (Phe), pyrene (Pyr) and benzo[a]pyrene (BaP) by measuring urinary monohydroxy-PAH (OH-PAH) derived from these parent compounds in habitual users of CCs, electronic cigarettes (ECs), heated tobacco products (HTPs), oral tobacco (OT), and nicotine replacement therapy products (NRTs). Non-users (NU) of any tobacco/nicotine products served as (negative) control group. Smokers exhibited the highest levels for all PAH biomarkers measured, almost all of which were significantly different from the NU and user groups of all other products investigated. CC smokers were the only group which showed a significant relationship between almost all PAH biomarkers and dose markers such as daily consumption, urinary nicotine equivalents (Nequ) and plasma cotinine (CotP). The ratios in urinary OH-PAH between CC and all other groups were dependent on the biomarker and range from < 2 to > 10. These ratios could at least partly be explained by the enzymes involved, their region-selectivity and inducibility by smoking. In conclusion, cigarette smokers (CC) were the only group, which showed product use dependent exposure to PAHs, whereas users of EC, HTP, NRT and OT were not distinguishable from NU of any tobacco/nicotine products.

Competitive Metabolism of Polycyclic Aromatic Hydrocarbons (PAHs): An Assessment Using In Vitro Metabolism and Physiologically Based Pharmacokinetic (PBPK) Modeling.

Humans are routinely exposed to complex mixtures such as polycyclic aromatic hydrocarbons (PAHs) rather than to single compounds, as are often assessed for hazards. Cytochrome P450 enzymes (CYPs) metabolize PAHs, and multiple PAHs found in mixtures can compete as substrates for individual CYPs (e.g., CYP1A1, CYP1B1, etc.). The objective of this study was to assess competitive inhibition of metabolism of PAH mixtures in humans and evaluate a key assumption of the Relative Potency Factor approach that common human exposures will not cause interactions among mixture components. To test this objective, we co-incubated binary mixtures of benzo[a]pyrene (BaP) and dibenzo[def,p]chrysene (DBC) in human hepatic microsomes and measured rates of enzymatic BaP and DBC disappearance. We observed competitive inhibition of BaP and DBC metabolism and measured inhibition coefficients (Ki), observing that BaP inhibited DBC metabolism more potently than DBC inhibited BaP metabolism (0.061 vs. 0.44 µM Ki, respectively). We developed a physiologically based pharmacokinetic (PBPK) interaction model by integrating PBPK models of DBC and BaP and incorporating measured metabolism inhibition coefficients. The PBPK model predicts significant increases in BaP and DBC concentrations in blood AUCs following high oral doses of PAHs (≥100 mg), five orders of magnitude higher than typical human exposures. We also measured inhibition coefficients of Supermix-10, a mixture of the most abundant PAHs measured at the Portland Harbor Superfund Site, on BaP and DBC metabolism. We observed similar potencies of inhibition coefficients of Supermix-10 compared to BaP and DBC. Overall, results of this study demonstrate that these PAHs compete for the same enzymes and, at high doses, inhibit metabolism and alter internal dosimetry of exposed PAHs. This approach predicts that BaP and DBC exposures required to observe metabolic interaction are much higher than typical human exposures, consistent with assumptions used when applying the Relative Potency Factor approach for PAH mixture risk assessment.

Characterization of benzo[a]pyrene and colchicine based on an in vivo repeat-dosing multi-endpoint genotoxicity quantitative assessment platform.

Two prototypical genotoxicants, benzo[a]pyrene (B[a]P) and colchicine (COL), were selected as model compounds to deduce their quantitative genotoxic dose-response relationship at low doses in a multi-endpoint genotoxicity assessment platform. Male Sprague-Dawley rats were treated with B[a]P (2.5-80 mg/kg bw/day) and COL (0.125-2 mg/kg bw/day) daily for 28 days. The parameters included were as follows: comet assay in the peripheral blood and liver, Pig-a gene mutation assay in the peripheral blood, and micronucleus test in the peripheral blood and bone marrow. A significant increase was observed in Pig-a mutant frequency in peripheral blood for B[a]P (started at 40 mg/kg bw/day on Day 14, started at 20 mg/kg bw/day on Day 28), whereas no statistical difference for COL was observed. Micronucleus frequency in reticulocytes of the peripheral blood and bone marrow increased significantly for B[a]P (80 mg/kg bw/day on Day 4, started at 20 mg/kg bw/day on Days 14 and 28 in the blood; started at 20 mg/kg bw/day on Day 28 in the bone marrow) and COL (started at 2 mg/kg bw/day on Day 14, 1 mg/kg bw/day on Day 28 in the blood; started at 1 mg/kg bw/day on Day 28 in the bone marrow). No statistical variation was found in indexes of comet assay at all time points for B[a]P and COL in the peripheral blood and liver. The dose-response relationships of Pig-a and micronucleus test data were analyzed for possible point of departures using three quantitative approaches, i.e., the benchmark dose, breakpoint dose, and no observed genotoxic effect level. The practical thresholds of the genotoxicity of B[a]P and COL estimated in this study were 0.122 and 0.0431 mg/kg bw/day, respectively, and our results also provided distinct genotoxic mode of action of the two chemicals.

Assessment of antitumor activity of BP-C1, a platinum-based anticancer agent with a lignin-derived polymeric ligand, in autochthonous induced and spontaneous carcinogenesis rodent models.

BACKGROUND: A standard approach to study the anticancer activity of novel drugs is their testing in animals with inoculated tumors, which has some limitations. An alternative is the use of spontaneous or carcinogen-induced tumor models as they have better translation potential. The carcinogen-induced and transgenic tumor models were used to assess the antitumor activity of BP-C1, a platinum-containing drug with lignin-derived polymeric ligand. METHODS: We used female Swiss-H-derived mice and Wistar female rats to induce autochthonous tumors via exposure to benzo[a]pyrene and 1,2-dimethylhydrazine, respectively. Additionally, transgenic HER-2/neu FVB/N female mice, prone to the development of spontaneous mammary carcinomas, were used. RESULTS: Antitumor activity of BP-C1 was observed in soft tissue sarcomas, induced by benzo[a]pyrene. The animals treated with BP-C1 exhibited more stabilizations and therapy responses compared to placebo controls. The efficacy of BP-C1 was somewhat reduced compared to cyclophosphamide; however, their combination resulted in an enhanced antitumor effect. For the 1,2-dimethylhydrazine-induced rat colon cancer model, BP-C1 reduced tumor multiplicity by 21-41 %. For mammary adenocarcinomas in HER-2/neu FVB/N mice, short-termed complete responses were observed in the BP-C1 groups with a frequency of 12-13 %, while complete responses were absent in the placebo group. CONCLUSION: The results acquired indicated a wide spectrum of antitumor activity of BP-C1.

Assessment of Alkylated and Unsubstituted Polycyclic Aromatic Hydrocarbons in Air in Urban and Semi-Urban Areas in Toronto, Canada.

22 alkylated polycyclic aromatic hydrocarbons (alk-PAHs) were characterized in ambient air individually for the first time in urban and semi-urban locations in Toronto, Canada. Five unsubstituted PAHs were included for comparison. Results from the measurements were used to estimate benzo[a]pyrene equivalent toxicity (BaPeq) of individual compounds in order to investigate the significance of a single compound in contributing to the overall toxic equivalency (TEQ) of air mixtures. To determine which compounds merit further investigation, BaPeq values of individual compounds were compared to the measured BaP toxicity. Our results showed that both unsubstituted and alkylated PAHs were more abundant in the urban area (38 and 30%, respectively). Benzo[a]pyrene levels at the urban location exceeded Ontario's 24 h guideline (40% of the events), and on average, it was 5 times higher than that at the semi-urban area. Gas-phase two- and three-ring compounds contributed up to 39% (urban) and 76% (semi-urban) of the TEQ of all compounds analyzed. Some alk-PAHs such as 7,12-dimethylbenzo[a]anthracene had a huge impact on the toxicity of urban air, and its BaPeq was on average 8 times higher than that of BaP. We emphasize that the toxic impact of alkylated and gaseous PAHs, which is not routinely included in many air monitoring programs, is significant and should not be neglected.

Urinary monohydroxylated polycyclic aromatic hydrocarbons in the general population from 26 provincial capital cities in China: Levels, influencing factors, and health risks.

Polycyclic aromatic hydrocarbons (PAHs) derived from the incomplete combustion of organic materials are associated with adverse health effects. However, little is known about PAH exposure levels and their influencing factors on a large scale in developing countries. In this study, urinary monohydroxylated metabolites of PAHs (OH-PAHs), including the metabolites of naphthalene, fluorene, phenanthrene, pyrene, chrysene, and benzo[a]pyrene, were measured in 1154 samples in the general population nationwide from 26 provincial capitals in China. Concentrations of OH-PAHs ranged from 1.39 to 228 µg/L. OH-Nap, metabolite of naphthalene, was the predominant compound, accounting for 65.1% of totals. People in eastern, southwest and northeast China, such as Shanghai, Kunming, Nanning, and Changchun, suffered more PAH exposure than other regions which might associate with sampling time, living habits of the subjects, and the imbalance of economic development and energy consumption across regions. Urinary OH-PAH concentrations were associated with body mass index, gender, and age, and smoking was the main correlating factor. Inhalation and diet might be the main exposure route of human exposure to PAHs, especially for smokers by inhalation. Hazard indices showed that no subject was exposed to PAHs with potential non-carcinogenic risk. Furthermore, the carcinogenic risk was the most significant health effects, with almost all subjects having carcinogenic risk values higher than the acceptable level of 10-6. Naphthalene and phenanthrene were the main contributors. The results also suggested a possible relationship between PAH exposure and lung cancer in the Chinese population. This first nationwide study on human internal exposure to PAHs provides a large body of scientific information for governmental decision-making about associated human health and the prevention of human exposure to PAHs.

Profiling of seasonal variation in and cancer risk assessment of benzo(a)pyrene and heavy metals in drinking water from Kirkuk city, Iraq.

Water samples at 13 sites were analyzed to evaluate heavy metals (cobalt, lead, manganese, copper) and benzo(a)pyrene using 2 methods of analysis (high-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA) kits). The Lesser Zap River is the main tributary of the Tigris and is used as a main source of drinking water in Kirkuk city through the General Kirkuk project. Risk evaluation for benzo(a)pyrene and lead in water samples was accomplished by Monte Carlo simulation. The highest concentrations of B(a)P were recorded at sites S7 and S5, with levels of 0.192 and 0.122 µg L-1 detected by HPLC and ELISA, respectively. The WHO guidelines for benzo[a]pyrene in drinking water recommend 0.7 µg L -1, and none of the samples surpassed this level; moreover, B(a)P levels exceeded EPA standards in 2014 (0.01 µg L-1), particularly when the liquid-liquid extraction method with HPLC was used. Carcinogenic risks for human adults and children exist and are highest during the rainy season as compared with the carcinogenic risk during the dry season and risks for children exceed those of adults. This indicates that the 2nd round of sampling (winter season) harbors more carcinogenic risk than the 1st round of sampling (dry season).

HepG2 spheroids as a biosensor-like cell-based system for (geno)toxicity assessment.

3D spheroids developed from HepG2 cells were used as a biosensor-like system for the detection of (geno)toxic effects induced by chemicals. Benzo(a)pyrene (B(a)P) and amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) with well-known mechanisms of action were used for system validation. HepG2 spheroids grown for 3 days were exposed to BaP and PhIP for 24 and 72 h. The growth and viability of spheroids were monitored by planimetry and Live/Dead staining of cells. Multi-parametric flow cytometric analysis was applied for simultaneous detection of specific end-effects including cell cycle analysis (Hoechst staining), cell proliferation (KI67 marker), and DNA double-strand breaks (ℽH2AX) induced by genotoxic compounds. Depending on the exposure concentration/time, BaP reduced spheroid growth, affected cell proliferation by arresting cells in S and G2 phase and induced DNA double-strand breaks (DSB). Simultaneous staining of ℽH2AX formation and cell cycle analysis revealed that after BaP (10 µM; 24 h) exposure 60% of cells in G0/G1 phase had DNA DSB, while after 72 h only 20% of cells contained DSB indicating efficient repair of DNA lesions. PhIP did not influence the spheroid size whereas accumulation of cells in the G2 phase occurred after both treatment times. The evaluation of DNA damage revealed that at 200 µM PhIP 50% of cells in G0/G1 phase had DNA DSB, which after 72-h exposure dropped to 40%, showing lower repair capacity of PhIP-induced DSB compared to BaP-induced. The developed approach using simultaneous detection of several parameters provides mechanistic data and thus contributes to more reliable genotoxicity assessment of chemicals as a high-content screening tool.

Occurrence of polycyclic aromatic hydrocarbons in human diet - exposure and risk assessment to consumer health.

BACKGROUND: Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants, they are also present in food, in which their presence results from environmental pollution and food processing processes. Many compounds from this group, such as benzo(a)pyrene show important toxicity, including genotoxic carcinogenicity. In food heavier PAHs significantly toxic are observed. OBJECTIVE: The aim of the study was assessment of consumers exposure to PAHs from the diet of surveyed respondents. The assessment of contaminants content in daily food rations is characterized by less uncertainty factor than the assessment based on data on the contamination of individual foodstuffs and their consumption by humans. MATERIAL AND METHODS: Research material consisted of daily diets obtained from respondents participating in the study. Content of 22 PAHs (fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo(c)fluorene, benz(a)anthracene, chrysene, 5-methylchrysene, perylene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(j)fluoranthene, benzo(e) pyrene, benzo(a)pyrene, benzo(ghi)perylene, indeno(1,2,3-cd)pyrene, dibenzo(a,h)anthracene, dibenzo(a,e)pyrene, dibenzo(a,l)pyrene, dibenzo(a,h)pyrene, dibenzo(a,i)pyrene) in each of diets was tested using liquid chromatography with a fluorescence detector. The samples were purified by saponification, size exclusion chromatography (SEC) and solid phase extraction (SPE). RESULTS: 52 respondents (n=52) took part in the study. The highest median of PAHs were found for pyrene (1.412 µg/kg), phenantrene (1.276 µg/kg), fluorene (1.151 µg/kg) and fluoranthene (1.087 µg/kg), they were about 10-80 higher than the levels of heavier PAHs. In group of heavy PAHs quantitatively prevailed benzo(e)pyrene (0.109 µg/kg), benzo(b) fluroanthene (0.070 µg/kg), benzo(ghi)perylene (0.065 µg/kg) and perylene (0.059 µg/kg). Generally the median level of contamination with light PAHs was 6.045 µg/kg, while with heavy ones 0.504 µg/kg, in the case of the sum of 4 PAHs regulated in EU law content was 0.301 µg/kg. In the tested samples average 24% of the PAH content was pyrene, light PAHs with a lower toxicity potential accounted for 92% of the content of tested compounds. Sum of 4 regulated PAHs accounted for 58% of content compounds selected by the EU as significant for the assessment of food contamination by PAHs. The composition of the participants' diets was analyzed in terms of determining factors influencing on high levels of PAHs. They were high fat level and presence of smoked or grilled meat and fish products. The mean exposure to benzo(a)pyrene was 0.52 ng/kg b.w. per day, while for the sum of 4 PAHs 3.29 ng/ kg b.w. per day. For light PAHs high exposure was 90.6 ng/kg b.w. per day, while for heavy PAH it was 10.7 ng/kg b.w. per day. Risk assessment was performed by calculating the value of margin of exposure (MoE), which for benzo(a)pyrene and for sum of 4 PAHs were above 25,000 in both considered: mean and high exposure scenario. CONCLUSIONS: Studied diets were a source of exposure to PAHs. Higher levels have been reported for light, less toxic PAH as compared to heavy PAH. In both considered scenarios margin of exposure were >25 000. In case of studied diets no risk for consumer was found.