Characterization of DNA and deoxyribonucleoside adducts formation by phenanthrene-9,10-oxide

The DNA adducts formed by phenanthrene-9,10-oxide (PhO) were analyzed using high performance liquid chromatography (HPLC) and monoclonal antibody based enzyme linked immunosorbent assay (ELISA) techniques. Polydeoxyribonucleotides polydG-polydC and polydA-polydT, were analyzed for comparison. The isocratic methanol-water HPLC chromatograms of the adducts of PhO with each of the four deoxyribonucleosides produced very similar HPLC retention times. In contrast, the MeOH and water gradient HPLC chromatograms of the PhO + dN adducts showed reasonably distinctive retention times. The HPLC chromatograms of PhO + DNA showed that there were ten presumed DNA adducts formed by PhO; these adducts were separated by HPLC and collected manually. The values for the 50% antibody inhibition point (I$\sb{50}$) for seven of the ten presumed DNA adducts were approximated in a competitive ELISA. The ELISA results indicated that the antibodies raised against thiol adducts of PhO cross react with PhO-DNA adducts. (Abstract shortened with permission of author.).