The measurement of oxidative damage to DNA by HPLC and GC/MS techniques.
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.
Halliwell
B
B
Division of Pulmonary/Critical Care Medicine, UC Davis Medical Center, Sacramento 95817.
Dizdaroglu
M
M
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002851:Chromatography, High Pressure Liquid; D004249:DNA Damage / Q000502:physiology*; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D005609:Free Radicals; D008401:Gas Chromatography-Mass Spectrometry; D006878:Hydroxides; D017665:Hydroxyl Radical; D010084:Oxidation-Reduction
D005609:Free Radicals; D006878:Hydroxides; D017665:Hydroxyl Radical; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine