Comparison of different methods of measuring 8-oxoguanine as a marker of oxidative DNA damage

Andrew Collins, J. Brown, M. Bogdanov, J. Cadet, M. Cooke, T. Douki, C. Dunster, J. Eakins, B. Epe, M. Evans, P. Farmer, C. M. Gedik, B. Halliwell, K. Herbert, T. Hofer, R. Hutchinson, A. Jenner, G. D.D. Jones, H. Kasai, F. KellyA. Lloret, S. Loft, J. Lunec, M. McEwan, L. Möller, R. Olinski, I. Podmore, H. Poulsen, J. L. Ravanat, J. E. Rees, F. Reetz, H. Shertzer, B. Spiegelhalder, R. Turesky, R. Tyrrell, J. Viña, D. Vinicombe, A. Weimann, B. de Wergifosse, S. G. Wood

Research output: Contribution to journalArticle

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Abstract

We are attempting to resolve some of the problems encountered in measuring 8-hydroxy-2'-deoxyguanosine (8-oxodG) in human cellular DNA as a marker of oxidative stress. Samples of authentic 8-oxodG were distributed, and participating laboratories undertook to analyse this material within a specified period. Most HPLC procedures gave values for 8-oxodG within ±40% of the target, as did two of four GC-MS procedures, and both LC-MS-MS methods. Calf thymus DNA samples containing increasing amounts of 8-oxodG were also distributed for analysis. Fewer than half the procedures tested were able to detect the dose response; those that were successful tended to be procedures with low coefficients of variation. For the analysis of 8-oxodG in human cells, where it is likely to be present at much lower concentrations than in the calf thymus DNA, it is crucial to reduce analytical variation to a minimum; a coefficient of variation of less than 10% should be the aim, to give reasonable precision. HPLC with amperometric electrochemical detection is not recommended, as it is less sensitive than coulometric detection. Immunological detection, 32P-postlabelling and LC-MS-MS are alternative approaches to measurement of 8-oxodG in DNA that, on the grounds of precision and detection of dose response, cannot at present be recommended.

Original languageEnglish (US)
Pages (from-to)333-341
Number of pages9
JournalFree Radical Research
Volume32
Issue number4
DOIs
StatePublished - Jan 1 2000

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Keywords

  • 8-hydroxy-2'-deoxyguanosine
  • GC-MS
  • HPLC
  • Methods validation
  • Oxidative DNA damage

Cite this

Collins, A., Brown, J., Bogdanov, M., Cadet, J., Cooke, M., Douki, T., Dunster, C., Eakins, J., Epe, B., Evans, M., Farmer, P., Gedik, C. M., Halliwell, B., Herbert, K., Hofer, T., Hutchinson, R., Jenner, A., Jones, G. D. D., Kasai, H., ... Wood, S. G. (2000). Comparison of different methods of measuring 8-oxoguanine as a marker of oxidative DNA damage. Free Radical Research, 32(4), 333-341. https://doi.org/10.1080/10715760000300331