Determination of in Vitro Formed DNA Adducts of 2-Amino-1-methyl-6-phenylimidazo[4,5-6]pyridine Using Capillary Liquid Chromatography/Electrospray Ionization/Tandem Mass Spectrometry

Diane Rindgen, Robert J. Turesky, Paul Vouros

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On-line reversed-phase capillary liquid chromatography/tandem mass spectrometry with electrospray ionization was used for the detection of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) adducts of DNA and compared to analysis by 32P-postlabeling. Structural information was obtained on low nanogram levels of a synthetic N-(deoxyguanosin-8-yl)-PhIP (dG-C8-PhIP) standard using collision-induced dissociation, and low picogram level detection was achieved for the targeted dG-C8-PhIP adduct using the selective multiple reaction monitoring scanning mode. The method was applied to the analysis of an in vitro reaction mixture identifying the dG-C8-PhIP adduct and providing evidence for the presence of two additional PhIP-modified deoxyguanosine adducts. The results were in qualitative agreement with those obtained by the 32P-postlabeling method. In addition, the mass spectrometric results revealed the occurrence of an unexpected product in the in vitro reaction mixture presumably resulting from cleavage through the guanine base.

Original languageEnglish (US)
Pages (from-to)1005-1013
Number of pages9
JournalChemical Research in Toxicology
Issue number8
StatePublished - Dec 1995


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