Fluorescence HPLC methods for detecting benzo[a]pyrene-7,8-dihydrodiol 9,10-oxide-deoxyadenosine adducts in...

The fluorescence of mononucleoside adducts derived from the binding of anti-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo [a]pyrene (BPDE I) to N6-deoxyadenosine (BPDE-dA adducts) is 10-100 times stronger (depending on the methanol/water solvent composition) than the fluorescence of adducts derived from the binding of this diol epoxide derivative to N2-deoxyguanosine. It is shown here that these fluorescence characteristics can be used to quantitate the relatively low yields of BPDE-dA adducts by fluorescence detection when BPDE-modified DNA is subjected to enzymatic degradation to the mononucleoside levels, followed by HPLC analysis of the digests.