Cancer Risk Assessment of Airborne PAHs Based on in Vitro Mixture Potency Factors

Dreij, K.; Mattsson, Å.; Jarvis, I.W.H.; Lim, H.; Hurkmans, J.; Gustafsson, J.; Bergvall, C.; Westerholm, R.; Johansson, C.; Stenius, U.
2017 | Environ. Sci. Technol. | 51 (8805-8814)

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 Chk1 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) lung 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.

Physical and chemical properties of RME biodiesel exhaust particles without engine modifications

Nyström, R.; Sadiktsis, I.; Ahmed, T.M.; Westerholm, R.; Koegler, J.H.; Blomberg, A.; Sandström, T.; Boman, C.
2016 | Fuel | 186 (261-269)

A major contributor to ambient particulate air pollution is exhaust from diesel engines and other vehicles, which can be linked to different adverse health effects. During the last decades, a global drive towards finding sustainable and clean bio-based alternative fuels for the transport sector has taken place and biodiesel is one of the most established alternatives today. To better assess the overall effects on a public health level when introducing biodiesel and other renewable fuels, a better understanding of the detailed exhaust particle properties, is needed. In this work, the physical and chemical properties of biodiesel exhaust particles were studied in comparison to standard diesel exhaust emissions, in an existing engine without modifications, focusing on particulate carbonaceous matter and PAH/Oxy-PAH as well as fine particle size distribution. An older off-road engine, produced between 1996 and 2004, was used with three different fuels/fuel blends; (1) 100 wt% low-sulfur standard petro diesel (SD), (2) 100 wt% rapeseed methyl ester biodiesel (B100) and (3) a blended fuel – B30 consisting of 30 wt% RME and 70 wt% SD. The study focused mainly on emissions from transient engine operation, but includes also idling conditions. The gaseous emissions measured for the biodiesel fuel were in general in accordance with previous reported data in the literature, and compared to the standard petro diesel the emissions of CO was lower while NOx emissions increased. The particulate mass concentration during transient operation was almost halved compared to when petro diesel was used and this was associated with a decrease in average particle size. The shift in particle mass and size was associated with a higher fraction of organic matter in general, considerable less PAH’s but a relative higher fraction of Oxy-PAH’s, when shifting from petro diesel to biodiesel.

PAH exposure and relationship between buccal micronucleus cytome assay and urinary 1-hydroxypyrene levels among cashew nut roasting workers

S. Batistuzzo; M.O. Galvão; E.S. Duarte; J.J. Hoelzemann; J. Menezes Filho; I. Sadiktsis; R. Westerholm; K. Dreij
2016 | Toxicol. Lett. | 258, supplement (S223-S224)

52nd Congress of the European Societies of Toxicology (EUROTOX) | September 4, 2016 | Fibes Congress Center Seville, Spain

Suspect screening of OH-PAHs and non-target screening of other organic compounds in wood smoke particles using HR-Orbitrap-MS

2016 | Chemosphere | 163 (313-321)

Wood combustion has been shown to contribute significantly to emissions of polycyclic aromatic hydrocarbons and hydroxylated polycyclic aromatic hydrocarbons, compounds with toxic and carcinogenic properties. However, only a small number of hydroxylated polycyclic aromatic hydrocarbons have been determined in particles from wood combustion, usually compounds with available reference standards. In this present study, suspect and non-target screening strategies were applied to characterize the wood smoke particles from four different wood types and two combustion conditions with respect to hydroxylated polycyclic aromatic hydrocarbons and other organic compounds. In the suspect screening, 32 peaks corresponding to 12 monohydroxylated masses were tentatively identified by elemental composition assignments and matching of isotopic pattern and fragments. More than one structure was suggested for most of the measured masses. Statistical analysis was performed on the non-target screening data in order to single out significant peaks having intensities that depend on the wood type and/or combustion condition. Significant peaks were found in both negative and positive ionization modes, with unique peaks for each wood type and combustion condition, as well as a combination of both factors. Furthermore, structural elucidation of some peaks was done by comparing the spectra in the samples with spectra found in the spectral databases. Six compounds were tentatively identified in positive ionization mode, and 19 in negative ionization mode. The results in this present study demonstrate that there are significant overall differences in the chemistry of wood smoke particles that depends on both the wood type and the combustion condition used.

Determination of semi-volatile and particle-associated polycyclic aromatic hydrocarbons in Stockholm air with emphasis on the highly carcinogenic dibenzopyrene isomers.

Masala, S.; Lim, H.; Bergvall, C.; Johansson, C.; Westerholm, R.
2016 | Atmos. Environ., Part A | 140 (370-380)

Particulate hydroxy-PAH emissions from a residential wood log stove using different fuels and burning conditions

Rozanna Avagyan; Robin Nyström; Robert Lindgren; Christoffer Boman; Roger Westerholm
2016 | Atmos Environ | 140 (1-9)

Hydroxylated polycyclic aromatic hydrocarbons are oxidation products of polycyclic aromatic hydrocarbons, but have not been studied as extensively as polycyclic aromatic hydrocarbons. Several studies have however shown that hydroxylated polycyclic aromatic hydrocarbons have toxic and carcinogenic properties. They have been detected in air samples in semi urban areas and combustion is assumed to be the primary source of those compounds. To better understand the formation and occurrence of particulate hydroxylated polycyclic aromatic hydrocarbons from residential wood log stove combustion, 9 hydroxylated polycyclic aromatic hydrocarbons and 2 hydroxy biphenyls were quantified in particles generated from four different types of wood logs (birch, spruce, pine, aspen) and two different combustion conditions (nominal and high burn rate). A previously developed method utilizing liquid chromatography – photo ionization tandem mass spectrometry and pressurized liquid extraction was used. Polycyclic aromatic hydrocarbons were analyzed along with hydroxylated polycyclic aromatic hydrocarbons. The hydroxylated polycyclic aromatic hydrocarbon emissions varied significantly across different wood types and burning conditions; the highest emissions for nominal burn rate were from spruce and for high burn rate from pine burning. Emissions from nominal burn rate corresponded on average to 15% of the emissions from high burn rate, with average emissions of 218 μg/MJfuel and 32.5 μg/MJfuel for high burn rate and nominal burn rate, respectively. Emissions of the measured hydroxylated polycyclic aromatic hydrocarbons corresponded on average to 28% of polycyclic aromatic hydrocarbons emissions.

This study shows that wood combustion is a large emission source of hydroxylated polycyclic aromatic hydrocarbons and that not only combustion conditions, but also wood type influences the emissions of hydroxylated polycyclic aromatic hydrocarbons and polycyclic aromatic hydrocarbons. There are few studies that have determined hydroxylated polycyclic aromatic hydrocarbons in emissions from wood combustion, and it is therefore necessary to further investigate the formation, occurrence and distribution of these compounds as they are present in significant amounts in wood smoke particles.

Determination of hydroxylated polycyclic aromatic hydrocarbons by HPLC-photoionization tandem mass spectrometry in wood smoke particles and soil samples

Rozanna Avagyan; Robin Nyström; Christoffer Boman; Roger Westerholm
2015 | Anal Bioanal Chem | 407 (16) (4523-4534)

A simple and fast method for analysis of hydroxylated polycyclic aromatic hydrocarbons using pressurized liquid extraction and high performance liquid chromatography utilizing photoionization tandem mass spectrometry was developed. Simultaneous separation and determination of nine hydroxylated polycyclic aromatic hydrocarbons and two hydroxy biphenyls could be performed in negative mode with a run time of 12 min, including equilibration in 5 min. The calibration curves were in two concentration ranges; 1–50 ng/mL and 0.01–50 μg/mL, with coefficients of correlation R 2 > 0.997. The limits of detection and method quantification limits were in the range of 9–56 pg and 5–38 ng/g, respectively. A two-level full factorial experimental design was used for screening of conditions with the highest impact on the extraction. The extraction procedure was automated and suitable for a large number of samples. The extraction recoveries ranged from 70 to 102 % and the matrix effects were between 92 and 104 %. The overall method was demonstrated on wood smoke particles and soil samples with good analytical performance, and five OH-PAHs were determined in the concentration range of 0.19–210 μg/g. As far as we know, hydroxylated polycyclic aromatic hydrocarbons were determined in wood smoke and soil samples using photoionization mass spectrometry for the first time in this present study. Accordingly, this study shows that high performance liquid chromatography photoionization tandem mass spectrometry can be a good option for the determination of hydroxylated polycyclic aromatic hydrocarbons in complex environmental samples.

Effect of wood smoke exposure on vascular function and thrombus formation in healthy fire fighters

Amanda L Hunter; Jon Unosson; Jenny A Bosson; Jeremy P Langrish; Jamshid Pourazar; Jennifer B Raftis; Mark R Miller; Andrew J Lucking; Christoffer Boman; Robin Nyström; Kenneth Donaldson; Andrew D Flapan; Anoop SV Shah; Louis Pung; Ioannis Sadiktsis; Silvia Masala; Roger Westerholm; Thomas Sandström; Anders Blomberg; David E Newby; Nicholas L Mills
2014 | Part Fibre Toxicol | 11 (62)
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Background
Myocardial infarction is the leading cause of death in fire fighters and has been linked with exposure to air pollution and fire suppression duties. We therefore investigated the effects of wood smoke exposure on vascular vasomotor and fibrinolytic function, and thrombus formation in healthy fire fighters.

Methods
In a double-blind randomized cross-over study, 16 healthy male fire fighters were exposed to wood smoke (~1 mg/m3 particulate matter concentration) or filtered air for one hour during intermittent exercise. Arterial pressure and stiffness were measured before and immediately after exposure, and forearm blood flow was measured during intra-brachial infusion of endothelium-dependent and -independent vasodilators 4–6 hours after exposure. Thrombus formation was assessed using the ex vivo Badimon chamber at 2 hours, and platelet activation was measured using flow cytometry for up to 24 hours after the exposure.

Results
Compared to filtered air, exposure to wood smoke increased blood carboxyhaemoglobin concentrations (1.3% versus 0.8%; P < 0.001), but had no effect on arterial pressure, augmentation index or pulse wave velocity (P > 0.05 for all). Whilst there was a dose-dependent increase in forearm blood flow with each vasodilator (P < 0.01 for all), there were no differences in blood flow responses to acetylcholine, sodium nitroprusside or verapamil between exposures (P > 0.05 for all). Following exposure to wood smoke, vasodilatation to bradykinin increased (P = 0.003), but there was no effect on bradykinin-induced tissue-plasminogen activator release, thrombus area or markers of platelet activation (P > 0.05 for all).

Conclusions
Wood smoke exposure does not impair vascular vasomotor or fibrinolytic function, or increase thrombus formation in fire fighters. Acute cardiovascular events following fire suppression may be precipitated by exposure to other air pollutants or through other mechanisms, such as strenuous physical exertion and dehydration.

Class separation of lipids and polycyclic aromatic hydrocarbons in normal phase High Performance Liquid Chromatography – A prospect for analysis of aromatics in edible vegetable oils and biodiesel exhaust particulates

Olsson, P; Sadiktsis, I.; Holmbäck, J.; Westerholm, R.
2014 | J. Chromatogr. A | 1360 (39-46)

The retention characteristics of the major lipid components in biodiesels and edible oils as well as representative polycyclic aromatic compounds (PAHs) have been investigated on five different normal phase HPLC stationary phases, in order to optimize class separation for an automatized online HPLC cleanup of PAHs prior GC-MS analysis. By stepwise comparison of different hexane/MTBE compositions as mobile phases on cyano-, phenyl-, pentabromobenzyl-, nitrophenyl- and amino- modified silica columns, the capacity and selectivity factors for each analyte and column could be calculated. It was concluded that the most suitable column for backflush isolation of PAHs in biodiesel and edible oil matrices was the pentabromobenzyl-modified silica (PBB). A previously described online HPLC-GC-MS system using the PBB column was then evaluated by qualitative and quantitative analysis of a biodiesel exhaust particulate extract and a vegetable oil reference material. The GC-MS full scan analysis of the biodiesel particulate extract showed that the lipids had been removed from the sample and a fraction containing PAHs and oxygenated derivatives thereof had been isolated. Quantified mass fractions of PAHs of the reference material BCR-458 agreed well for most of the certified PAH mass fractions in the spiked coconut oil reference material.

Determination of oxygenated and native polycyclic aromatic hydrocarbons in urban dust and diesel particulate matter standard reference materials using pressurized liquid extraction and LC-GC/MS

T. Mohammad Ahmed, C. Bergvall, M. Åberg, R. Westerholm
2014 | Anal Bioanal Chem

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.

Pressurized liquid extraction as an alternative to the Soxhlet extraction procedure stated in the US EPA method TO-13A for the recovery of polycyclic aromatic hydrocarbons adsorbed on polyurethane foam plugs

S. Masala, Ulf Rannug, R. Westerholm
2014 | 6 (8420-8425)

The aim of the present study was to develop a pressurized liquid extraction (PLE) method as an alternative to the relatively time consuming Soxhlet extraction procedure described in the United States Environmental Protection Agency (US EPA) method TO-13A for the extraction of PAHs adsorbed onto polyurethane foam plugs (PUFs). For this purpose PUF air samples were collected and split into two parts: one part extracted using PLE and the other one using Soxhlet extraction. Comparable PAH concentrations were obtained upon analysis of the extracts showing that the PLE method developed in this work is a more convenient choice than the commonly used Soxhlet extraction technique proposed by US EPA for the determination of PAHs in air samples. In fact, the developed PLE method required shorter assay times (minutes versus hours), less solvent consumption and simpler operational methods. The exhaustiveness of the developed PLE method was evaluated using repeat static extraction cycles, demonstrating an extraction efficiency for the PAHs of greater than 99 %. The PLE method was then applied to diesel exhaust and wood smoke PUF samples showing an extraction efficiency for the PAHs of greater than 93 % and 96 %, respectively. Furthermore, a PLE method for PUF cleaning was developed as well and employed as an alternative to Soxhlet extraction. The PLE methods developed for cleaning and extracting PUFs presented in this work are suitable to be used in mutagenicity studies using the Ames Salmonella assay as no mutagenicity was found in the PLE generated blanks.

Tire tread wear particles in ambient air – a previously unknown source of human exposure to the biocide 2-mercaptobenzothiazole

2014 | Environ Sci Pollut Res Int | 21 (19) (11580-11586)

Urban particulate matter (PM), asphalt, and tire samples were investigated for their content of benzothiazole and benzothiazole derivates. The purpose of this study was to examine whether wear particles, i.e., tire tread wear or road surface wear, could contribute to atmospheric concentrations of benzothiazole derivatives. Airborne particulate matter (PM10) sampled at a busy street in Stockholm, Sweden, contained on average 17 pg/m3 benzothiazole and 64 pg/m3 2-mercaptobenzothiazole, and the total suspended particulate-associated benzothiazole and 2-mercaptobenzothiazole concentrations were 199 and 591 pg/m3, respectively. This indicates that tire tread wear may be a major source of these benzothiazoles to urban air PM in Stockholm. Furthermore, 2-mercaptobenzothiazole was determined in urban air particulates for the first time in this study, and its presence in inhalable PM10 implies that the human exposure to this biocide is underestimated. This calls for a revision of the risk assessments of 2-mercaptobenzothiazole exposure to humans which currently is limited to occupational exposure.

Contact information

Visiting addresses:

Geovetenskapens Hus,
Svante Arrhenius väg 8, Stockholm

Arrheniuslaboratoriet, Svante Arrhenius väg 16, Stockholm (Unit for Analytical and Toxicological Chemistry)

Mailing address:
Department of Environmental Science and Analytical Chemistry (ACES)
Stockholm University
106 91 Stockholm

Press enquiries should be directed to:

Stella Papadopoulou
Science Communicator
Phone +46 (0)8 674 70 11
stella.papadopoulou@aces.su.se