Separation of 32P-labeled 3′,5′-bisphosphate nucleotides of polycyclic aromatic hydrocarbon anti-diol-epoxides and derivatives

Abstract 32 P-Postlabeling-HPLC is a highly sensitive analytical method for identification of chemical-modified DNA adducts isolated from samples obtained from experimental animals or humans exposed to carcinogenic chemicals. To determine optimal 32 P-postlabeling-HPLC conditions for efficient separation, we report here the use of ten diol-epoxide-modified 3′,5′-bisphosphate deoxynucleotides derived from benzo[ a ]pyrene (BaP), nitrated BaP, and related compounds. After testing ODS-modified, C 4 -modified, phenyl-modified, diphenyl-modified, and cyclodextrin-bonded reversed-phase HPLC columns, we found that the Vydac diphenyl-modified column can efficiently separate these 3′,5′-bisphosphate deoxynucleotides. The results suggest that 32 P-postlabeling-HPLC is a potentially useful methodology for detecting environmental carcinogens that can be metabolized to diol-epoxides. The relationships between the structures of anti -diol-epoxides and HPLC retention order are also discussed.