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32 P-POSTLABELING AND HPLC SEPARATION OF DNA ADDUCTS FORMED BYDIESEL EXHAUST EXTRACTS IN VITRO AND IN MOUSE SKIN AND LUNG AFTERTOPICAL TREATMENT
Citation:
Savela, K., L. King, J. Gallagher, AND J. Lewtas. 32 P-POSTLABELING AND HPLC SEPARATION OF DNA ADDUCTS FORMED BYDIESEL EXHAUST EXTRACTS IN VITRO AND IN MOUSE SKIN AND LUNG AFTERTOPICAL TREATMENT. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-95/524, 1995.
Description:
Diesel exhaust extracts contain many carcinogenic compounds which have been shown to form PAH- and nitrated-PAH-DNA adducts in rodent skin and lung. he aim of this study was to characterize by "P-postlabeling, TLC and HPLC the primary postlabeled PAH-DNA adduct(s) formed in vitro and in vivo by diesel extracts. he diesel particle extracts had known concentrations of B(a)P, benzo(b,j,k)fluoranthenes and chrysene DNA adducts were analyzed in calf thymus DNA incubated in vitro with PAH activated by S9 mix and in skin and lung DNA from topically treated mice. he main diesel derived DNA adduct formed in vitro and in vivo did not co-migrate on HPLC and large TLC plates with anti-BPDE-, B(b)F-, BO)F-, B(k)F- or chrysene-standards. y cochromatography DNA adducts formed by chrysene from both in vitro and in vivo samples were identified. issan diesel extract containing higher PAH-concentrations than Volkswagen automobile extract formed skin DNA adducts that co-migrated with chrysene- and anti BPDE-DNA-derived adducts. e conclude that the use of highly sensitive 32P postlabeling method combined with HPLC improves the identification of PAH-adducts formed by complex mixtures such as diesel exhaust extracts.