SYNTHESIS AND P-32 POSTLABELING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY SEPARATION OF DIASTEREOMERIC 1,N-2-(1,3-PROPANO)-2'-DEOXYGUANOSINE 3'-PHOSPHATE ADDUCTS FORMED FROM 4-HYDROXY-2-NONENAL/

4-Hydroxy-8-nonenal (HNE), a major electrophilic byproduct of lipid pe roxidation, is mutagenic and cytotoxic. The two pairs of HNE-derived d iastereomeric 1,N-2-propanodeoxyguanosine 3'-monophosphate adducts wer e synthesized from reaction of HNE with 2'-deoxyguanosine 3'-monophosp hate. After HPLC separation, these adducts were characterized by UV-vi sible absorption and negative ion electrospray ionization MS/MS analys is. To further characterize the structures, these adducts were dephosp horylated to the corresponding HNE-modified deoxyguanosine adducts and their HPLC retention times and UV spectra were compared with those of the synthetic standards prepared from reaction of HNE with 2'-deoxygu anosine, Separation of these adducts by P-32-postlabeling/HPLC was dev eloped. Reaction of HNE with calf thymus DNA resulted in only one pair of diastereomeric adducts, with one adduct predominantly formed with a modification level of 1.2 +/- 0.5 adducts/10(7) nucleotides.