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Analysis of Urinary 8-oxo-7,8-dihydro-2-deoxyguanosine by Liquid Chromatography–Tandem Mass Spectrometry

  • Mark D. Evans
  • Rajinder Singh
  • Vilas Mistry
  • Peter B. Farmer
  • Marcus S. Cooke
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 610)

Abstract

The ability to non-invasively monitor DNA oxidation and its repair has significant utility in large-scale, population-based studies. Such studies could include assessments of the efficacy of antioxidant intervention strategies, pathological roles of DNA oxidation in various disease states and population or inter-individual differences in antioxidant defence and DNA repair. The analysis of urine, or indeed any extracellular matrix, for 8-oxo-7,8-dihydro-2ʹ-deoxyguanosine (8-oxodG), using chromatographic or immunoassay procedures, is by far the most popular method to non-invasively assess oxidative insult to the genome. The actual biological significance of the presence of extracellular 8-oxodG is still a subject for debate however. Studies are slowly ruling out confounding factors such as diet and cell turnover, which would leave endogenous processes, notably repair, as the sole source of extracellular 8-oxodG. The method described herein exploits the non-invasive properties of urine sampling, coupled with efficient extraction of 8-oxodG by a validated solid-phase extraction procedure. Subsequent analysis by liquid chromatography–tandem mass spectrometry has the advantages of sensitivity, internal standardisation and robust peak identification.

Key words

Oxidative stress DNA damage DNA repair chromatography solid-phase extraction urine liquid chromatography mass spectrometry 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Mark D. Evans
    • 1
  • Rajinder Singh
    • 2
  • Vilas Mistry
    • 3
  • Peter B. Farmer
    • 2
  • Marcus S. Cooke
    • 4
  1. 1.Radiation & Oxidative Stress Group Department of Cancer Studies and Molecular MedicineUniversity of LeicesterLeicesterUK
  2. 2.Cancer Biomarkers and Prevention Group, Department of Cancer Studies and Molecular MedicineUniversity of LeicesterLeicesterUK
  3. 3.Radiation & Oxidative Stress GroupUniversity of LeicesterLeicesterUK
  4. 4.Radiation & Oxidative Stress Group Department of GeneticsUniversity of LeicesterLeicesterUK

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