Automated method for the direct analysis of 8-oxo-guanosine and 8-oxo-2'-deoxyguanosine in human urine using ultraperformance liquid chromatography and tandem mass spectrometry

Free Radic Biol Med. 2009 Sep 1;47(5):629-35. doi: 10.1016/j.freeradbiomed.2009.06.002. Epub 2009 Jun 23.


The potential use of oxidative stress-induced DNA and RNA damage products as biomarkers is an important aspect of biomedical research. There is a need for assays with high specificity and sensitivity that also can be used in molecular epidemiology studies with a large number of subjects. In addition there is a need for assays that can measure more than one product from DNA oxidation. We present a sensitive, precise, and accurate method for quantitative analysis of the oxidized nucleosides 8-oxoGuo and 8-oxodG in human urine. The assay is based on automated sample handling using a BIOMEK 3000 Workstation, and UPLC-ESI(+)-MS/MS analysis. High specificity is evidenced by the use of qualifier ions for both analytes. The quantification limit in urine samples is 1 nM for both analytes. Accuracy and precision were documented, showing average recoveries of 106.2% (8-oxoGuo) and 106.9% (8-oxodG), and overall precision (within-day and between-days) of 6.1 and 4.4%, respectively.

Publication types

  • Validation Study

MeSH terms

  • 8-Hydroxy-2'-Deoxyguanosine
  • Automation / methods
  • Chromatography, Liquid / instrumentation
  • Chromatography, Liquid / methods
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / analysis
  • Deoxyguanosine / urine
  • Guanosine / analogs & derivatives*
  • Guanosine / analysis
  • Guanosine / urine
  • Humans
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Specimen Handling / methods
  • Tandem Mass Spectrometry / instrumentation*
  • Tandem Mass Spectrometry / methods*
  • Urinalysis / instrumentation
  • Urinalysis / methods*


  • Guanosine
  • 8-hydroxyguanosine
  • 8-Hydroxy-2'-Deoxyguanosine
  • Deoxyguanosine