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Simultaneous detection of five different 2-hydroxyethyl-DNA adducts formed by ethylene oxide exposure, using a high-performance liquid chromatography/electrospray ionisation tandem mass spectrometry assay

By Elaine M. Tompkins, Donald J. L. Jones, John H. Lamb, Debbie A. Marsden, Peter B. Farmer and Karen Brown

Abstract

This is the author's final draft of the version published as Rapid Communications in Mass Spectrometry, 2008, 22 (1),pp.19-28. The definitive version is available at www3.interscience.wiley.com, Doi: 10.1002/rcm.3328.A method has been developed for the simultaneous detection and quantitation of five different (2-hydroxyethyl)-DNA adducts (HE-DNA adducts) that could be formed as a result of exposure to ethylene oxide (EO). In addition to the major N7-HE-guanine (N7-HEG) adducts this assay can also measure the less prevalent but potentially more biologically significant N1-HE-2’-deoxyadenosine (N1-HEdA), O6-HE-2’-deoxyguanosine (O6-HEdG), N6-HE-2’-deoxyadenosine (N6-HEdA) and N3-HE-2’-deoxyuridine adducts (N3-HEdU). The method involves the isolation of HE-adducts from the unmodified nucleosides by either neutral thermal hydrolysis or enzymatic digestion, followed by HPLC purification, before detection and quantification by liquid-chromatography tandem mass spectrometry (LC-MS/MS) using selective reaction monitoring (SRM). The limits of detection were in the range 0.5 – 25 fmoles for each individual adduct, making this one of the most sensitive assays available for the detection of N7-HEG. To illustrate the possible applications of the assay, it has been employed in the measurement of endogenous/background and EO-induced HE-adducts in a variety of DNA samples

Topics: LC-MS/MS, SRM, 2-hydroxyethyl, ethylene oxide, DNA adducts
Publisher: John Wiley & Sons
Year: 2007
DOI identifier: 10.1002/rcm.3328
OAI identifier: oai:lra.le.ac.uk:2381/3653

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