Abstract
Evidence is accumulating for the predominant involvement of singlet oxygen ( 1 O 2 ) in the oxidation reactions of major cellular targets such as DNA and membranes on exposure to UVA radiation. In this chapter, emphasis is placed on the measurement of 8-oxodGuo 5, a relevant biomarker of 1 O 2 oxidation, in cellular DNA using improved chromatographic assays. In addition, a short survey of available information on 1 O 2 oxidation of the guanine moiety of DNA and model compounds is provided. Many efforts are currently devoted to the optimization of assays aimed at singling out the formation in cellular DNA of oxidized bases with emphasis on 8-oxoGua, a relevant DNA biomaker of oxidative stress. In this respect, three chromatographic methods involving HPLC-ECD, HPLC-MS/MS, and HPLC/GC-MS have been compared and critically evaluated. The measurement of oxidative base damage within cellular DNA is still a challenging analytical problem. It should be remembered that the requested limit of sensitivity of the applied method should be close to one modification per 10 6 to 10 7 DNA bases. In addition, the possibility of artifact formation, because of DNA oxidation during the workup, does not facilitate the achievement of such a goal.
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