Abstract
Sensitive and specific isotope dilution liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) methods were developed for the detection and quantitative analysis ofS-{2-(N7-guanyl)ethyl}glutathione as a DNA adduct formed upon exposure of animals to carcinogenic 1,2-dihaloethanes. Separation and analysis were performed using microbore HPLC coupled in-line to an electrospray ionization triple quadrupole mass spectrometer.S-{2-(N7-guanyl)[2H4]ethyl} glutathione was synthesized and used as internal standard. These methods provide structural confirmation of the adduct as well as quantitative analysis with the accuracy and precision necessary to measure biologically relevant levels in small tissue sample sizes (< 1 g). The sample detection limits inin vivotissue extracts were 100 pg and 5 pg on-column for LC/MS and LC/MS/MS methods, respectively. Selected-ion monitoring mode was used to monitor the product ions of the doubly charged molecular ion. The application of these methods was demonstrated by measuring the DNA adduct levels in rat and fish samples after exposure to 1,2-dihaloethanes. The method has application in studies of DNA adduct formation as a biological marker of exposure to carcinogens and for environmental monitoring of 1,2-dihaloethanes.
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