Utilização da tecnica HPLC/APCI-MS/MS para determinação e identificação de danos ao DNA

AUTOR(ES)
DATA DE PUBLICAÇÃO

2004

RESUMO

Reactive oxygen species produce oxidized bases, deoxyribose lesions and DNA strand breaks in mammalian cells. Free radicaIs are produced in cells by cellular metabolism and by exogenous agents. These species react with biomolecules in cells, inc1udingDNA. The resulting damage to DNA, which is also called oxidative damage to DNA, is implicated in mutagenesis, carcinogenesis, and aging. We introduced a new methodology using Atmosphere Pressure Chemical Ionization (APCI) and HPLC/tandem mass spectrometry (HPLC/APCI-MS/MS). This technique for the measurement of modified nuc1eosides can simultaneously measure numerous products (8-hydroxyguanine, dihydrothymine, 4,6-diamino-5-(formamido)pyrimidine, isobarbituric acid (5-hydroxyuracil), 5-hydroxymethyluracil and thymine glycol), and provide positive identification and accurate quantification for monitoring in MRM mode. The responses for all damage bases and the four normal bases were highly linear with correlation coefficients 0.9950-0.9999 and detection limit infmols. We applied the method for quantification and identification ofthese bases in cultured PC-12 cells, a rat pheochromocytoma (fibroblasts) cellline irradiated for 2,4 and 6 hours following FL exposure

ASSUNTO(S)

doença de estresse oxidativo alzheimer espectrometria de massa

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