CHEMILUMINESCENT DETECTION OF THYMINE HYDROPEROXIDES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
High-performance liquid chromatography (HPLC) with chemiluminescence detection was used to detect thymine hydroperoxides, which are expected to occur in biological tissues as primary products in free radical-mediated DNA damage. The thymine hydroperoxides were chemically prepared by the acid-catalysed hydrogen peroxide oxidation of thymine and 5-hydroxymethyluracil and their hydroperoxide products were identified as trans- or cis-5-chloro-6-hydroperoxy-5,6-dihydrothymine and 5-hydroperoxymethyluracil, respectively, by means of NMR, IR and mass spectrometry. The results confirmed that these thymine hydroperoxides art specifically detectable at picomole levels by the described HPLC method with high reproducibility.
Journal Of Chromatography