The extractable organic material (EOM) from atmospheric total suspended particles (TSP) contains several organic compounds including non-substituted polycyclic aromatic hydrocarbons (PAHs), alkyl-PAHs, and nitro-PAHs. These chemicals seem to be among the key drivers of TSP genotoxicity. We have shown previously that the mutagenic potencies of the EOM from Limeira, Stockholm, and Kyoto, cities with markedly different meteorological conditions and pollution sources are similar. Here we compare the profiles of non-substituted PAHs (27 congeners), alkyl-PAHs (15 congeners), and nitro-PAHs (7 congeners) from the same EOM samples from these cities. We also compared the genotoxicity profiles using comet and micronucleus assays in human bronchial epithelial cells. The profiles of PAHs, as well as the cytotoxic and genotoxic potencies when expressed in EOM, were quite similar among the studied cities. It seems that despite the differences in meteorological conditions and pollution sources of the cities, removal, mixing, and different atmospheric transformation processes may be contributing to the similarity of the PAHs composition and genotoxicity profiles. More studies are required to verify if this would be a general rule applicable to other cities. Although these profiles were similar for all three cities, the EOM concentration in the atmospheres is markedly different. Thus, the population of Limeira (similar to 10-fold more EOM/m(3) than Stockholm and similar to 6-fold more than Kyoto) is exposed to higher concentrations of genotoxic pollutants, and Kyoto's population is 1.5-fold more exposed than Stockholm's. Therefore, to reduce the risk of human exposure to TSP genotoxins, the volume of emissions needs to be reduced.

Atmospheric particulate matter (PM) organic fractions from urban centers are frequently mutagenic for the Salmonella/microsome assay. This mutagenicity is related to both primary and secondary pollutants, and meteorological conditions have great influence on the secondary pollutant's formation. Our objective was to compare the mutagenicity of atmospheric total suspended particulates (TSP) from three cities with marked different meteorological conditions and TSP concentrations: Limeira (Brazil) with 99.0 mu g/m(3), Stockholm (Sweden) with 6.2 mu g/m(3), and Kyoto (Japan) with 28.0 mu g/m(3). For comparison, we used the same batch of filters, sample extraction method, and Salmonella/microsome testing protocol with 11 strains of Salmonella with and without metabolic activation. Samples were collected during winter and pooled into one single extract representing each city. All samples were mutagenic for all tested strains, except for TA102. Based on the strain's selectivity, nitroarenes, polycyclic aromatic hydrocarbons, and aromatic amines play a predominant role in the mutagenicity of these samples. The mutagenic potencies expressed by mass of extracted organic material (EOM; revertants/mu g EOM) were similar (similar to twofold difference) among the cities, despite differences in meteorological conditions and pollution sources. In contrast, the mutagenic potencies expressed by air volume (rev/m(3)) varied similar to 20-fold, with Limeira > Kyoto approximate to Stockholm. These results are the first systematic assessment of air mutagenicity from cities on three continents using the same protocols. The results confirm that the mutagenic potency expressed by EOM mass is similar regardless of continent of origin, whereas the mutagenic potency expressed by air volume can vary by orders of magnitude. Environ. Mol. Mutagen. 2019.

Emission factors for oxygenated polycyclic aromatic hydrocarbons (OPAHs) and PAHs have been determined from two different fuel flexible light duty vehicles operated at -7 degrees C in the New European Driving Cycle (NEDC) and at +22 degrees C in the Artemis Driving Cycle (ADC). Three different gasoline/ethanol blends, commercially available in Sweden, were tested i.e., gasoline E5, with 5% v/v ethanol and ethanol fuel E85 with 85% v/v ethanol and winter time quality E70 with 70% v/v ethanol, respectively. The results showed greatly increased emissions of both OPAHs and PAHs at cold engine start conditions (-7 degrees C in the NEDC) compared to warm engine start (+ 22 degrees C in the ADC). For the OPAHs, higher average total emission factors were obtained when running on E85 compared to E5 at both cold 2.72 mu g/km vs 1.11 mu g/km and warm 0.19 mu g/km vs 0.11 mu g/km starting conditions with the highest emissions when using E70 at -7 degrees C 4.12 mu g/km. The same trend was found for the PAHs at cold engine start with higher average total emission factors when using ethanol fuel 71.5 mu g/km and 60.0 mu g/km for E70 and E85, respectively compared to gasoline E5 (20.2 mu g/km). Slightly higher average total PAH emissions were obtained when operating at + 22 degrees C with E5 compared to with E85 1.23 mu g/km vs 0.72 mu g/km.

An online two-dimensional (2D) liquid chromatography/2D gas chromatography system with two mass-selective detectors has been developed on the basis of a previous system with two flame ionization detectors. The method translation involved the change of carrier gas from hydrogen to helium, column dimension and detectors. The 2D system with two mass-selective detectors was validated with use of polycyclic aromatic hydrocarbon (PAH) standards and two standard reference materials from air and diesel exhaust. Furthermore, the system was applied to a real sample, wood smoke particulates. The PAH values determined correlated well with the previous data and those from the National Institute of Standards and Technology. The system enhanced the benefits of the previous system, which were limited by the low detectability and lack of mass selectivity. This study shows an automated 2D system that is valid for PAH analysis of complex environmental samples directly from crude extracts.

Complex mixtures of polycyclic aromatic hydrocarbons (PAHs) are common environmental pollutants associated with adverse human health effects including cancer. However, the risk of exposure to mixtures is difficult to estimate, and risk assessment by whole mixture potency evaluations has been suggested. To facilitate this, reliable in vitro based testing systems are necessary. Here, we investigated if activation of DNA damage signaling in vitro could be an endpoint for developing whole mixture potency factors (MPFs) for airborne PAHs. Activation of DNA damage signaling was assessed by phosphorylation of Chid and H2AX using Western blotting. To validate the in vitro approach, potency factors were determined for seven individual PAHs which were in very good agreement with established potency factors based on cancer data in vivo. Applying the method using Stockholm air PAH samples indicated MPFs with orders of magnitude higher carcinogenic potency than predicted by established in vivo-based potency factors. Applying the MPFs in cancer risk assessment suggested that 45.4 (6% of all) cancer cases per year in Stockholm are due to airborne PAHs. Applying established models resulted in <1 cancer case per year, which is far from expected levels. We conclude that our in vitro based approach for establishing MPFs could be a novel method to assess whole mixture samples of airborne PAHs to improve health risk assessment.

The concentrations of polycyclic aromatic hydrocarbons (PAHs) have been determined in the gaseous phase and in various particulate matter (PM) size fractions at different locations in and outside of Stockholm, Sweden, representative of street level, urban and rural background. The focus has been on the seldom determined but highly carcinogenic dibenzopyrene isomers (DBPs) dibenzo[a,I]pyrene, dibenzo [a,e]pyrene, dibenzo[a,i]pyrene and dibenzo[a,h]pyrene. PAHs with 3 rings were found to be mainly associated with the vapor phase (>90%) whereas PAHs with 5-6 rings were mostly associated with particulate matter (>92%) and the 4-ringed PAHs partitioned between the two phases. PAH abundance was determined to be in the order street level > urban background > rural background with the PM10 street level 2010 mean of benzo[a]pyrene (B[a]P) reaching 0.24 ng/m(3), well below the EU annual limit value of 1 ng/m(3). In addition, higher PAH concentrations were found in the sub-micron particle fraction (PM1) as compared to the super -micron fraction (PM1-10) with the abundance in PM1 varying between 57 and 86% of the total PAHs. The B[a]P equivalent concentrations derived for DB[a,l]P and total DBPs exceeded 1-2 and 2-4 times, respectively, that of B[a]P at the four sampling sites; therefore underestimation of the cancer risk posed by PAHs in air could be made if the DBPs were not considered in risk assessment using the toxic equivalency approach, whilst the high correlation (p < 0.001) found in the relative concentrations supports the use of B[a]P as a marker substance for assessment of the carcinogenic risk associated to PAHs. However, the big difference in concentration ratios of B[a]P and the DBPs between the present study and some literature data calls for further research to evaluate the temporal and spatial invariance of the B[a]P/DBP ratios.

Background: Smoke from combustion of biomass fuels is a major risk factor for respiratory disease, but the underlying mechanisms are poorly understood. The aim of this study was to determine whether exposure to wood smoke from incomplete combustion would elicit airway inflammation in humans. Methods: Fourteen healthy subjects underwent controlled exposures on two separate occasions to filtered air and wood smoke from incomplete combustion with PM1 concentration at 314 mu g/m(3) for 3 h in a chamber. Bronchoscopy with bronchial wash (BW), bronchoalveolar lavage (BAL) and endobronchial mucosal biopsies was performed after 24 h. Differential cell counts and soluble components were analyzed, with biopsies stained for inflammatory markers using immunohistochemistry. In parallel experiments, the toxicity of the particulate matter (PM) generated during the chamber exposures was investigated in vitro using the RAW264.7 macrophage cell line. Results: Significant reductions in macrophage, neutrophil and lymphocyte numbers were observed in BW (p < 0.01, < 0.05, < 0.05, respectively) following the wood smoke exposure, with a reduction in lymphocytes numbers in BAL fluid (< 0.01. This unexpected cellular response was accompanied by decreased levels of sICAM-1, MPO and MMP-9 (p < 0.05, < 0.05 and < 0.01). In contrast, significant increases in submucosal and epithelial CD3+ cells, epithelial CD8+ cells and submucosal mast cells (p < 0.01, < 0.05, < 0.05 and < 0.05, respectively), were observed after wood smoke exposure. The in vitro data demonstrated that wood smoke particles generated under these incomplete combustion conditions induced cell death and DNA damage, with only minor inflammatory responses. Conclusions: Short-term exposure to sooty PAH rich wood smoke did not induce an acute neutrophilic inflammation, a classic hallmark of air pollution exposure in humans. While minor proinflammatory lymphocytic and mast cells effects were observed in the bronchial biopsies, significant reductions in BW and BAL cells and soluble components were noted. This unexpected observation, combined with the in vitro data, suggests that wood smoke particles from incomplete combustion could be potentially cytotoxic. Additional research is required to establish the mechanism of this dramatic reduction in airway leukocytes and to clarify how this acute response contributes to the adverse health effects attributed to wood smoke exposure.

Benz[j]aceanthrylene (B[j]A) is a cyclopenta-fused polycyclic aromatic hydrocarbon with strong mutagenic and carcinogenic effects. We have identified B[j]A in air particulate matter (PM) in samples collected in Stockholm, Sweden and in Limeira, Brazil using LC-GC/MS analysis. Determined concentrations ranged between 1.57 and 12.7 and 19.6-30.2 pg/m(3) in Stockholm and Limeira, respectively, which was 11-30 times less than benzo[a]pyrene (B[a]P) concentrations. Activation of the DNA damage response was evaluated after exposure to B[j]A in HepG2 cells in comparison to B[a]P. We found that significantly lower concentrations of B[j]A were needed for an effect on cell viability compared to B[a]P, and equimolar exposure resulted in significant more DNA damage with B[j]A. Additionally, levels of gamma H2AX, pChk1, p53, pp53, and p21 proteins were higher in response to B[j]A than B[a]P. On the basis of dose response induction of pChk1 and gamma H2AX, B[j]A potency was 12.5- and 33.3-fold higher than B[a]P, respectively. Although B[j]A levels in air were low, including B[j]A in the estimation of excess lifetime cancer risk increased the risk up to 2-fold depending on which potency factor for B[j]A was applied. Together, our results show that B[j]A could be an important contributor to the cancer risk of air PM.

The objective of this study was to develop a novel analytical chemistry method, comprised of a coupled high-performance liquid chromatography-gas chromatography/mass spectrometry system (LC-GC/MS) with low detection limits and high selectivity, for the identification and determination of oxygenated polycyclic aromatic hydrocarbons (OPAHs) and polycyclic aromatic hydrocarbons (PAHs) in urban air and diesel particulate matter. The linear range of the four OPAHs, which include 9,10-anthraquinone, 4H-cyclopenta[def]phenanthrene-4-one, benzanthrone, and 7,12-benz[a]anthraquinone, was 0.7 pg-43.3 ng with limits of detection (LODs) and limits of quantification (LOQs) on the order of 0.2-0.8 and 0.7-1.3 pg, respectively. The LODs in this study are generally lower than values reported in the literature, which can be explained by using large-volume injection. The recoveries of the OPAHs spiked onto glass fiber filters using two different pressurized liquid extraction (PLE) methods were in the ranges of 84-107 and 67-110 %, respectively. The analytical protocols were validated using the following National Institute of Standards and Technology standard reference materials: SRM 1649a (Urban Dust), SRM 1650b (Diesel Particulate Matter), and SRM 2975 (Diesel Particulate Matter, Industrial Forklift). The measured mass fractions of the OPAHs in the standard reference materials (SRMs) in this present study are higher than the values from the literature, except for benzanthrone in SRM 1649a (Urban Dust). In addition to the OPAHs, 44 PAHs could be detected and quantified from the same particulate extract used in this protocol. Using data from the literature and applying a two-sided t test at the 5 % level using Bonferroni correction, significant differences were found between the tested PLE methods for individual PAHs. However, the measured mass fractions of the PAHs were comparable, similar to, or higher than those previously reported in the literature.

Black carbon containing emissions from biomass combustion are being transformed in the atmosphere upon processing induced by tropospheric ozone and UV. The knowledge today is very limited on how atmospheric processing affects the toxicological properties of the emissions. The aim of this study was to investigate the influence of ozone initiated (dark) atmospheric processing on the physicochemical and toxicological properties of particulate emissions from wood combustion. Emissions from a conventional wood stove operated at two combustion conditions (nominal and hot air starved) were diluted and transferred to a chamber. Particulate matter (PM) was collected before and after ozone addition to the chamber using an impactor. Detailed chemical and physical characterization was performed on chamber air and collected PM. The collected PM was investigated toxicologically in vitro with a mouse macrophage model, endpoints included: cell cycle analysis, viability, inflammation and genotoxicity. The results suggest that changes in the organic fraction, including polycyclic aromatic hydrocarbons (PAHs) are the main driver for differences in obtained toxicological effects. Fresh hot air starved emissions containing a higher organic and PAH mass-fraction affected cell viability stronger than fresh emissions from nominal combustion. The PAH mass fractions decreased upon aging due to chemical degradation. Dark aging increased genotoxicity, reduced viability and reduced release of inflammatory markers. These differences were statistically significant for single doses and typically less pronounced. We hypothesize that the alterations in toxicity upon simulated dark aging in the atmosphere may be caused by reaction products that form when PAHs and other organic compounds react with ozone and nitrate radicals. (C) 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).