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

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  • A.R. Collins
  • C.M. Gedik
  • S. Wood
  • A. White
  • J. Bubois
  • P. Duez
  • J.F. Rees
  • R. Legall
  • L. Degand
  • A. Jensen
  • H. Poulsen
  • A. Weimann
  • B.R. Jensen
  • J. Cadet
  • Thierry Douki
  • J.L. Ravanat
  • H. Faure
  • M. Tripier
  • I. Morel
  • O. 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. Duracková
  • L. Hlinchikova
  • P. Korytar
  • M. Dusinska
  • C. Mislanova
  • J. Vina
  • A. Lloret
  • L. Möller
  • T. Hofer
  • E. Gremaud
  • L. Fay
  • R. Stadler
  • J. Eakins
  • F. Progna
  • J. O`Brien
  • R. Elliott
  • S.B. Astley
  • A. Bailley
  • K. Herbert
  • D. Chauhan
  • F. Kelly
  • C. Dunster
  • J. Lunec
  • I. Podmore
  • P. Patel
  • S. Johnson
  • M. Evans
  • A. White
  • R. Tyrrell
  • Matthew Gordon
  • Chris Wild
  • Laura Hardie
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 CVof 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.
OriginalsprogEngelsk
TidsskriftFree Radical Research
Udgave nummervol. 36
Sider (fra-til)239-245
ISSN1071-5762
DOI
StatusUdgivet - 2002

Bibliografisk note

Paper id:: 12071341

ID: 139535