Isotope dilution high-performance liquid chromatography–electrospray tandem mass spectrometry assay for the measurement of 8-oxo-7,8-dihydro-2′-deoxyguanosine in biological samples |
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| Authors: | J-L Ravanat B Duretz A Guiller T Douki J Cadet |
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| Institution: | aDépartement de Recherche Fondamentale sur la Matière Condensée /SCIB/Laboratoire `Lésions des Acides Nucléiques', CEA-Grenoble, 17, Avenue des Martyrs, F-38054 Grenoble Cedex 9, France;bPerkin-Elmer Applied Biosystems, 25 Av de la Baltique BP 96, 91943 Courtaboeuf Cedex, France |
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| Abstract: | A sensitive and specific assay aimed at measuring 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo) has been developed by associating a reversed-phase liquid chromatographic separation with an electrospray tandem mass spectrometric detection. The HPLC–MS approach in the single ion monitoring (SIM) mode and the HPLC–MS/MS assay in the multiple reaction monitoring (MRM) mode have been compared, using isotopically labeled M+4] 8-oxodGuo as the internal standard. The limit of detection of 8-oxodGuo was found to be around 5 pmol and 20 fmol for the HPLC–MS and HPLC–MS/MS methods, respectively. The HPLC–MS/MS assay is sensitive enough to allow the determination of the level of 8-oxodGuo in cellular liver DNA and in urine samples. |
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| Keywords: | DNA biomarkers Oxidative stress 8-Oxo-7 8-dihydro-2&prime -deoxyguanosine |
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