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Genotoxic potential of organic extracts from particle emissions of diesel and rapeseed oil powered engines
Authors:Topinka Jan  Milcova Alena  Schmuczerova Jana  Mazac Martin  Pechout Martin  Vojtisek-Lom Michal
Institution:Institute of Experimental Medicine AS CR, Vídeňská 1083, 14220 Prague 4, Czech Republic. jtopinka@biomed.cas.cz
Abstract:The present study was performed to identify possible genotoxicity induced by organic extracts from particulate matter in the exhaust of two typical diesel engines run on diesel fuel and neat heated fuel-grade rapeseed oil: a Cummins ISBe4 engine tested using the World Harmonized Steady State Test Cycle (WHSC) and modified Engine Steady Cycle (ESC) and a Zetor 1505 engine tested using the Non-Road Steady State Cycle (NRSC). In addition, biodiesel B-100 (neat methylester of rapeseed oil) was tested in the Cummins engine run on the modified ESC. Diluted exhaust was sampled with high-volume samplers on Teflon coated filters. Filters were extracted with dichlormethane (DCM) and DNA adduct levels induced by extractable organic matter (EOM) in an acellular assay of calf thymus DNA coupled with (32)P-postlabeling in the presence and absence of rat liver microsomal S9 fraction were employed. Simultaneously, the chemical analysis of 12 priority PAHs in EOM, including 7 carcinogenic PAHs (c-PAHs) was performed. The results suggest that diesel emissions contain substantially more total PAHs than rapeseed oil emissions (for the ESC) or that these concentrations were comparable (for the WHSC and NRSC), while c-PAHs levels were comparable (for the ESC) or significantly higher (for the WHSC and NRSC) for rapeseed oil emissions. DNA adduct levels induced by diesel and rapeseed oil derived EOM were comparable, but consistently slightly higher for diesel than for rapeseed oil. Highly significant correlations were found between 12 priority PAHs concentrations and DNA adduct levels (0.980; p<0.001) and these correlations were even stronger for c-PAHs (0.990; p<0.001). Metabolic activation by the microsomal S9 fraction resulted in several fold higher genotoxicity, suggesting a major contribution of PAHs to genotoxicity. Directly acting compounds, other than c-PAHs, and not requiring S9 to exhibit DNA reactivity were also significant. Generally, DNA adduct levels were more dependent on the type of engine and the test cycle than on the fuel. Our findings suggest that the genotoxicity of particulate emissions from the combustion of rapeseed oil is significant and is comparable to that from the combustion of diesel fuel. A more detailed study is ongoing to verify and extent these preliminary findings.
Keywords:ANT  athracene  B[a]P  benzo[a]pyrene  B[b]F  benzo[b]fluoranthene  B[k]F  benzo[k]fluoranthene  B[a]A  benz[a]anthracene  B[ghi]P  benzo[ghi]perylene  BPDE  benzo[a]pyrene-r-7  t-8-dihydrodiol-t-9  10-epoxide[±]  c-PAHs  carcinogenic polycyclic aromatic hydrocarbons  CHRY  chrysene  CT-DNA  calf thymus DNA  DRZ  diagonal radioactive zone  DB[ah]A  dibenz[ah]anthracene  DCM  dichlormethane  ESC  Engine Steady State Cycle  EOM  extractable organic matter  HPLC  high performance liquid chromatography  FLU  fluoranthene  I[cd]P  indeno[cd]pyrene  NRSC  Non-Road Steady State Cycle  PM  particulate matter  PHE  phenanthrene  PYR  pyrene  RAL  relative adduct labeling  SDS  sodium dodecyl sulfate  TEF  toxic equivalency factor  TEQ  toxicity equivalent  TLC  thin layer chromatography  WHSC  World Harmonized Steady State Test Cycle
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