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

Reference:

Collins, A. R., Gedik, C., Wood, S., White, A., Dubois, J., Duez, P., Rees, J. F., Legall, R., Degand, L., Loft, S., Jensen, A., Poulsen, H., Weimann, A., Jensen, B. R., Cadet, J., Douki, T., Ravanat, J. L., Faure, H., Tripier, M., Morel, I., Sergent, O., Cillard, P., Morin, B., Epe, B., Phoa, N., Hartwig, A., Pelzer, A., Dolara, P., Casalini, C., Guglielmi, F., Luceri, C., Kasai, H., Kido, R., Olinski, R., Bialkowski, K., Durackova, Z., Hlincikova, L., Korytar, P., Dusinska, M., Mislanova, C., Vina, J., Lloret, A., Moller, L., Hofer, T., Gremaud, E., Fay, L., Stadler, R., Eakins, J., Pognan, F., O'Brien, J., Elliott, R., Astley, S., Bailley, A., Herbert, K., Chauhan, D., Kelly, F., Dunster, C., Lunec, J., Podmore, I., Patel, P., Johnson, S., Evans, M., Tyrrell, R., Gordon, M., Wild, C., Hardie, L., Smith, E. and Escodd, [N. V.], 2002. Inter-laboratory validation of procedures for measuring 8-oxo-7,8-dihydroguanine/8-oxo-7,8-dihydro-2 '-deoxyguanosine in DNA. Free Radical Research, 36 (3), pp. 239-245.

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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.

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