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Analysis of oxidative and free radical induced DNA damage
Váňová, Nela ; Kučera, Radim (advisor) ; Kovaříková, Petra (referee)
Analysis of oxidative and free radical induced DNA damage Diploma thesis Nela Váňová Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Free radicals and reactive oxygen species (ROS) are highly reactive molecules capable of modifications of biomolecules, including DNA. 5',8-cyclopurine-2'deoxynucleosides represent a group of DNA lesions characterized by concomitant damage to both sugar and base moieties of the same purine nucleoside that are together with 8-oxo-2'-deoxypurines among the major lesions formed by attack of free radicals (e.g. hydroxyl radical). Quantification of oxidative and free radical induced DNA lesions as biomarkers of oxidative stress has a high importance in study of their role in human health and disease. For quantification of these DNA lesions in gamma irradiated samples, high performance liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) will be utilized. Before injection into the LC/MS/MS, irradiated samples, treated by enzymatic digestion in order to gain free nucleosides, have to be desalted and DNA lesions have to be separated from undamaged nucleosides. A new HPLC/UV method was developed for separation of (5'R)-5',8-cyclo-2'- deoxyadenosine;...
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