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

Andrew R. Collins, Catherine Gedik, Sharon Wood, Ann White, Jacques Dubois, Pierre Duez, Jean Fraņois Rees, Rozenn Legall, Liliane Degand, Steffen Loft, Annie Jensen, Henrik Poulsen, Allan Weimann, Bente Riis Jensen, Jean Cadet, Thierry Douki, Jean Luc Ravanat, Henry Faure, Michèle Tripier, Isabelle MorelOdile Sergent, Pierre Cillard, Bénédicte Morin, Bernd Epe, Nicole Phoa, Andrea Hartwig, Anke Pelzer, Piero Dolara, Chiara Casalini, Francesco Guglielmi, Cristina Luceri, Hiroshi Kasai, Rie Kido, Ryszard Olinski, Karol Bialkowski, Zdena Duracková, Lucia Hlinciková, Peter Korytar, Mária Dusinská, Csilla Mislanová, José Viña, Ana Lloret, Lennart Möller, Tim Hofer, Eric Gremaud, Laurent Fay, Richard Stadler, Julie Eakins, Fraņois Pognan, John O'Brien, Ruan Elliott, Siân Astley, Angela Bailley, Karl Herbert, Dina Chauhan, Frank Kelly, Chrisi Dunster, Joseph Lunec, Ian Podmore, Parul Patel, Samantha Johnson, Mark Evans, Andrea White, Rex Tyrrell, Matthew Gordon, Chris Wild, Laura Hardie, Elizabeth Smith

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76 Scopus citations

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
StatePublished - 1 Mar 2002
Externally publishedYes

Keywords

  • 8-Oxodexyguanosine (8-oxo-dG)
  • Methods validation
  • Oxidative DNA damage
  • Photosensitiser

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