Synthesis and analysis of DNA adducts of arylamines

Arylamines and nitroarenes are very important environmental and occupational pollutants. Genotoxic effects of arylamines are believed to be initiated by the formation of DNA adducts. DNA adducts of arylamines have been found in experimental animals and in exposed humans, and are predominantly formed with the carbon 8 of 2'-deoxyguanosine. Reference standards are necessary to develop methods for the quantification of DNA-adducts. Therefore, we have synthesized the 2'-deoxyguanosin-8-yl adducts of 2-methylaniline, 2-chloroaniline, 4-chloroaniline, 2,4-dimethylaniline, and 2,6-dimethylaniline. The products were characterized by H-1-NMR, C-13-NMR, MS and UV. The corresponding 2'-deoxyguanosine-3'-monophosphate adducts were synthesized for the quantification of DNA adducts by the P-32-postlabelling technique. A GC-MS method was developed for the analysis of the new adducts as an alternative to the P-32-postlabelling. DNA was spiked with the synthesized adducts and treated with 0.3 M NaOH overnight at 110 degrees C in the presence of a deuterated internal standard. We observed up to 80% recovery from about 1 adduct in 10(8) to 1 in 10(5) nucleotides.