Diesel engine exhaust emission fractions: clastogenic effects in vitro
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Despite being hailed as a green fuel, emissions from diesel engines including particulate matter (PM10 and PM 2.5) have been implicated in a range of adverse human health effects from lung and bladder cancers to premature mortality. In this study diesel engine exhaust emissions were collected from a light duty direct injection diesel engine on a standard test bed. Engine conditions of speed and load were altered to provide a set of total emission samples from over the engine's operating range. Diesel emission samples collected were fractionated on a silica column into aliphatic, aromatic, and polar groups of compounds, which were tested for their genotoxicity in the chromosome aberration assay in Chinese hamster ovary CHO-K1 cells both with and without metabolic activation (rat liver S9 fraction). The aliphatic fractions did not exhibit cytotoxicity up to the maximum concentration assayed, and one emission sample (3000 rpm speed and 5 Nm load) assayed for effect on chromosome aberrations was not clastogenic (up to 600 µg/ml). The aromatic fractions of all engine emission samples assayed and of the fuel were not clastogenic, but did show high levels of cytotoxicity at relatively low doses, raising concern that any genotoxic effect was masked by the toxicity of certain chemicals within the fraction. Further fractionation, using HPLC, was therefore performed which separated the aromatics into various ring sizes. Assay of the ring fractions showed evidence of increasing clastogenicity with increasing ring size, with the 3+ -ring fractions of both the fuel and one emission sample clearly clastogenic when assayed with metabolic activation (evidence of the presence of indirect-acting genotoxic compounds within both samples). The final fractions to be assayed, the polar fractions, were clastogenic when assayed both with and without metabolic activation. All seven fractions from emission samples collected over a range of speed and load conditions caused highly significant increases in chromosome aberrations at concentrations as low as 20 µg/ml. An engine running for less than 30 minutes at 1000 rpm speed and 55 Nm load (urban driving conditions for a heavily laden vehicle) would emit 148 mg of polar group compounds for every litre of fuel consumed. Polar compounds have been shown to be a highly mutagenic fraction of air particulate samples, and as diesel emissions contribute up to 80 % of the particulate matter in urban air in some areas, diesel emissions and the polar compounds in particular are of real concern to human health.
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