DETERMINATION OF IN-VITRO FORMED DNA-ADDUCTS OF 2-AMINO-1-METHYL-6-PHENYLIMIDAZO[4,5-B]PYRIDINE USING CAPILLARY LIQUID-CHROMATOGRAPHY ELECTROSPRAY-IONIZATION TANDEM MASS-SPECTROMETRY

On-line reversed-phase capillary liquid chromatography/tandem mass spe ctrometry with electrospray ionization was used for the detection of 2 -amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) adducts of DNA a nd compared to analysis by P-32-postlabeling. Structural information w as obtained on low nanogram levels of a synthetic N-(deoxyguanosin-8-y l)-PhIP (dG-C8-PhIP) standard using collision-induced dissociation, an d low picogram level detection was achieved for the targeted dG-C8-PhI P adduct using the selective multiple reaction monitoring scanning mod e. The method was applied to the analysis of an in vitro reaction mixt ure identifying the dG-C8-PhIP adduct and providing evidence for the p resence of two additional PhIP-modified deoxyguanosine adducts. The re sults were in qualitative agreement with those obtained by the P-32-po stlabeling method. In addition, the mass spectrometric results reveale d the occurrence of an unexpected product in the in vitro reaction mix ture presumably resulting from cleavage through the guanine base.