Measurement of oxidized bases in DNA. Comparison between HPLC-EC sand GC-MS assays

J. L. Ravanat, T. Douki, R. Turesky, J. Cadet

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Abstract

The detection of oxidized bases in cellular DNA is a challenging problem due to the sensitivity required that should be high enough in order to detect one modification per 106 DNA bases. In this respect, gas chromatography mass spectrometry (GC-MS) has been extensively used during the last ten years. However, comparison of the results obtained by this method and those inferred from high performance liquid chromatography analyses coupled with an electrochemical detection (HPLC-EC) indicated an overestimation in the level of the oxidized DNA bases in the GC-MS measurement. A comparative evaluation of the two analytical methods allows us to demonstrate that this overestimation is due to an artefactual oxidation of normal DNA bases during the derivatization reaction that is required prior to CC-MS analysis. Purification of the modified DNA bases prior to the derivatization reaction prevents the artefact to occur. Under these conditions, the results obtained using the CC-MS method and the HPLC-EC assay are similar.

Original languageEnglish (US)
Pages (from-to)306-312
Number of pages7
JournalJournal de Chimie Physique et de Physico-Chimie Biologique
Volume94
Issue number2
DOIs
StatePublished - Jan 1 1997

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Keywords

  • DNA damage
  • GC-MS
  • HPLC-EC
  • Oxidized DNA bases

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