Chlorine substitution promotes phenyl radical loss from C8-phenoxy-2'-deoxyguanosine adducts: implications for biomarker identification from chlorophenol exposure.

Chlorophenols are persistent organic pollutants, which undergo peroxidase-mediated oxidation to afford phenolic radical intermediates that react at the C8-site of 2'-deoxyguanosine (dG) to generate oxygen-linked C8-dG adducts. Such adducts are expected to contribute to chlorophenol toxicity and serve as effective dose biomarkers for chlorophenol exposure. Electrospray ionization mass spectrometry (ESI-MS) was employed to study collision induced dissociation (CID) for a family of such phenolic O-linked C8-dG adducts. Fragmentation of the deprotonated nucleosides demonstrates that an unexpected homolytic cleavage of the ether linkage to release phenyl radicals and a nucleoside distonic ion with m/z 281 competes effectively with commonly observed breakage of the glycosidic bond to release the deprotonated nucleobase. Increased chlorination of the phenyl ring enhances phenyl radical loss. Density functional theory calculations demonstrate that Cl-substitution decreases phenyl radical stability but promotes homolytic breakage of the C8-phenyl bond in the C8-dG adduct. The calculations suggest that phenyl radical loss is driven by destabilizing steric (electrostatic repulsion) interactions between the ether oxygen atom and ortho-chlorines on the phenyl ring. The distonic ion at m/z 281 represents a unique dissociation product for deprotonated O-linked C8-dG adducts and may prove useful for selective detection of relevant biomarkers for chlorophenol exposure by tandem mass spectrometry using selective reaction monitoring.