Genotoxicity screening using biocatalyst/DNA films and capillary LC-MS/MS.
Detection of DNA adducts can serve as a basis for genotoxicty screening of new chemicals and drugs. We report here a simple, sensitive procedure for this purpose using films containing DNA and a biocatalyst to mimic the metabolic action of human liver cytochrome P450s. DNA adducts formed from an in-situ-generated toxic metabolite (styrene oxide) were detected at subpicomole levels after neutral thermal hydrolysis of the DNA films and analysis with capillary liquid chromatography with on-line column preconcentration and MS/MS detection. An on-line column switching system allowed for increased sample loading volume and analyte preconcentration. This approach provides an estimate of the relative rate of DNA damage.
Tarun
Maricar
M
Bajrami
Besnik
B
Rusling
James F
JF
D002853:Chromatography, Liquid; D004247:DNA / Q000737:chemistry*; D018736:DNA Adducts / Q000032:analysis*; D004852:Epoxy Compounds / Q000737:chemistry; D006801:Humans; D009152:Mutagenicity Tests / Q000379:methods*; D009211:Myoglobin / Q000737:chemistry; D053719:Tandem Mass Spectrometry
D018736:DNA Adducts; D004852:Epoxy Compounds; D009211:Myoglobin; D004247:DNA; C013690:styrene oxide