Comparison of different methods of measuring 8-oxoguanine as a marker of oxidative DNA damage
- 1 January 2000
- journal article
- research article
- Published by Informa UK Limited in Free Radical Research
- Vol. 32 (4), 333-341
- https://doi.org/10.1080/10715760000300331
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.Keywords
This publication has 4 references indexed in Scilit:
- Controlled oxidation of calf thymus DNA to produce standard samples for 8-oxodeoxyguanosine analysis; effects of freeze-drying, storage and hydrolysis conditions.Free Radical Research, 2000
- ESCODD: European standards committee on oxidative DNA damageFree Radical Research, 1998
- Measuring oxidative damage to DNA; HPLC and the comet assay compared.Free Radical Research, 1998
- Problems in the measurement of 8-oxoguanine in human DNA. Report of a workshop, DNA oxidation, held in Aberdeen, UK, 19-21 January, 1997Carcinogenesis: Integrative Cancer Research, 1997