The measurement of oxidative damage to DNA by HPLC and GC/MS techniques

Free Radic Res Commun. 1992;16(2):75-87. doi: 10.3109/10715769209049161.

Abstract

Oxidative damage to DNA has been measured by quantitating 8-hydroxy-2'-deoxyguanosine (8-OHdGuo) after enzymic digestion of DNA, followed by HPLC separation and electrochemical detection. Alternatively, 8-hydroxyguanine (and a wide range of other base-derived products of free radical attack) may be measured after acidic hydrolysis of DNA or chromatin, followed by derivatization and gas-chromatography/mass spectrometry. Both techniques have comparable sensitivity, but GC/MS enables determination of a wide variety of chemical changes to all four DNA bases and it can be applied to DNA-protein complexes. However, the two techniques do not always give similar results. Potential reasons for this are discussed. Greater attention to methodological questions is required before using measurement of 8-OHdGuo as a "routine" marker of oxidative DNA damage in vivo.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Review

MeSH terms

  • 8-Hydroxy-2'-Deoxyguanosine
  • Chromatography, High Pressure Liquid
  • DNA Damage / physiology*
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / metabolism
  • Free Radicals
  • Gas Chromatography-Mass Spectrometry
  • Hydroxides
  • Hydroxyl Radical
  • Oxidation-Reduction

Substances

  • Free Radicals
  • Hydroxides
  • Hydroxyl Radical
  • 8-Hydroxy-2'-Deoxyguanosine
  • Deoxyguanosine