Determination of cis-thymine glycol in DNA by gas chromatography-mass spectrometry with selected ion recording and multiple reaction monitoring.
A novel method for the determination of cis-thymine glycol in DNA has been developed, using gas chromatography-mass spectrometry with selected ion recording or multiple reaction monitoring. The procedure involves acidic hydrolysis of DNA in the presence of the internal standard cis-[2H3]thymine glycol, followed by derivatisation with N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide. The method was validated on DNA that had been oxidatively modified in vitro by radiation treatment, and was then applied to determine cis-thymine glycol in human placental DNA. Background levels of 5.45+/-2.98 ng cis-thymine glycol/mg DNA were observed in the human samples.