Analysis of urinary 8-oxo-7,8-dihydro-purine-2'-deoxyribonucleosides by LC-MS/MS and improved ELISA

Free Radic Res. 2008 Oct;42(10):831-40. doi: 10.1080/10715760802506323.

Abstract

Non-invasive monitoring of oxidative stress is highly desirable. Urinary 7,8-8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is a biologically relevant and convenient analytical target. However, immunoassays can over-estimate levels of urinary 8-oxodG. Measurement of more than one DNA oxidation product in urine would be advantageous in terms of mechanistic information. Urines samples were analysed for 8-oxodG by solid-phase extraction/LC-MS/MS and ELISA. The solid-phase extraction/LC-MS/MS assay was also applied to the analysis of urinary 7,8-dihydro-8-oxo-2'-deoxyadenosine (8-oxodA). Concurring with previous reports, urinary 8-oxodG measured by ELISA was significantly higher than levels measured by LC-MS/MS. However, apparent improvement in the specificity of the commercially available Japanese Institute for the Control of Ageing (JaICA) ELISA brought mean LC-MS/MS and ELISA measurements of urinary 8-oxodG into agreement. Urinary 8-oxodA was undetectable in all urines, despite efficient recovery by solid phase extraction. Exploitation of the advantages of ELISA may be enhanced by a simple modification to the assay procedure, although chromatographic techniques still remain the 'gold standard' techniques for analysis of urinary 8-oxodG. Urinary 8-oxodA is either not present or below the limit of detection of the instrumentation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 8-Hydroxy-2'-Deoxyguanosine
  • Adult
  • Biomarkers / urine
  • Chromatography, Liquid*
  • DNA Damage*
  • Deoxyadenosines / urine*
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / urine
  • Enzyme-Linked Immunosorbent Assay*
  • Female
  • Humans
  • Male
  • Oxidative Stress*
  • Reproducibility of Results
  • Tandem Mass Spectrometry*
  • Young Adult

Substances

  • Biomarkers
  • Deoxyadenosines
  • 2'-deoxy-7,8-dihydro-8-oxoadenosine
  • 8-Hydroxy-2'-Deoxyguanosine
  • Deoxyguanosine