Measurement of formamidopyrimidines in DNA

Published

December 15, 2008

Author(s)

Pawel Jaruga, Guldal Kirkali, Miral M. Dizdar

Abstract

Formamidopyrimidines, 4,6-diamino-5-formamidopyrimidine (FapyAde) and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua), are among major lesions in DNA generated by hydroxyl radical attack, UV radiation or photosensitization in vitro and in vivo. FapyAde and FapyGua exist in living cells at detectable background levels and are formed by exposure of cells to DNA-damaging agents. Numerous prokaryotic and eukaryotic DNA glycosylases exist for the repair of formamidopyrimidines by base excision repair pathway in cells indicating their biological significance. Moreover, they are premutagenic lesions, albeit to different extents, revealing a possible role in disease processes. Methodologies using gas chromatography/mass spectrometry (GC/MS) with capillary columns have been developed to accurately measure FapyAde and FapyGua in DNA in vitro and in vivo. Stable isotope-labeled analogues of these compounds have been synthesized and are commercially available to be used as internal standards for quantification. GC/MS with isotope-dilution provides excellent sensitivity and selectivity for positive identification and accurate quantification, and has widely been applied in the past to the measurement of formamidopyrimidines under numerous experimental conditions. This article reports on the details of this GC/MS methodology.

Citation

Free Radical Biology and Medicine

Volume

45

Issue

12

Pub Type

Journals

Keywords

Formamidopyrimidines, DNA repair, GC/MS, Mutagenesis, Oxidative DNA damage

Citation

Jaruga, P. , Kirkali, G. and Dizdar, M. (2008), Measurement of formamidopyrimidines in DNA, Free Radical Biology and Medicine (Accessed December 4, 2024)