Inter-laboratory validation of procedures for measuring 8-oxo-7,8-dihydroguanine/8-oxo-7,8-dihydro-2 '-deoxyguanosine in DNA

A R Collins, C Gedik, S Wood, A White, J Dubois, P Duez, J F Rees, R Legall, L Degand, S Loft, A Jensen, H Poulsen, A Weimann, B R Jensen, J Cadet, T Douki, J L Ravanat, H Faure, M Tripier, I MorelO Sergent, P Cillard, B Morin, B Epe, N Phoa, A Hartwig, A Pelzer, P Dolara, C Casalini, F Guglielmi, C Luceri, H Kasai, R Kido, R Olinski, K Bialkowski, Z Durackova, L Hlincikova, P Korytar, M Dusinska, C Mislanova, J Vina, A Lloret, L Moller, T Hofer, E Gremaud, L Fay, R Stadler, J Eakins, F Pognan, J O'Brien, R Elliott, S Astley, A Bailley, K Herbert, D Chauhan, F Kelly, C Dunster, J Lunec, I Podmore, P Patel, S Johnson, M Evans, R Tyrrell, M Gordon, C Wild, L Hardie, E Smith, [No Value] Escodd

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Abstract

The aim of ESCODD, a European Commission funded Concerted Action, is to improve the precision and accuracy of methods for measuring 8-oxo-7,8-dihydroguanine (8-oxoGua) or the nucleoside (8-oxodG). On two occasions, participating laboratories received samples of different concentrations of 8-oxodG for analysis. About half the results returned (for 8-oxodG) were within 20% of the median values. Coefficients of variation (for three identical samples) were commonly around 10%. A sample of calf thymus DNA was sent, dry, to all laboratories. Analysis of 8-oxoGua/8-oxodG in this sample was a test of hydrolysis methods. Almost half the reported results were within 20% of the median value, and half obtained a CV of less than 10%. In order to test sensitivity, as well as precision, DNA was treated with photosensitiser and light to introduce increasing amounts of 8-oxoGua and samples were sent to members. Median values calculated from all returned results were 45.6 (untreated), 53.9, 60.4 and 65.6 8-oxoGua/10(6) Gua; only seven laboratories detected the increase over the whole range, while all but one detected a dose response over two concentration intervals. Results in this trial reflect a continuing improvement in precision and accuracy. The next challenge will be the analysis of 8-oxodG in DNA isolated from cells or tissue, where the concentration is much lower than in calf thymus DNA.
Original languageEnglish
Pages (from-to)239-245
Number of pages7
JournalFree Radical Research
Volume36
Issue number3
DOIs
Publication statusPublished - 2002

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