Quantitative measurement of DNA adducts using neutral hydrolysis and LC-MS. Validation of genotoxicity sensors.
Neutral hydrolysis and LC-MS/MS analysis of 6-nm-thick DNA-polyion films used in voltammetric genotoxicity screening sensors showed that concentrations of N7-guanine DNA adducts with methyl methanesulfonate and styrene oxide increased with incubation time with the same trends as found for sensor response. Results show that the genotoxicity sensors can be used to estimate relative DNA damage rates for chemical toxicity screening. Neutral thermal hydrolysis provided a relatively clean sample matrix allowing quantitative estimates of nucleobase adducts after several minutes of incubation with damage agents. In addition, an approximate standardization procedure for neutral thermal hydrolysis was developed and validated that avoids need for a pure standard and should be useful in cases where nucleobase adduct standards are unavailable or where their identities are unknown.
Tarun
Maricar
M
Department of Chemistry, 55 North Eagleville Road, University of Connecticut, Storrs, Connecticut 06269-3060, USA.
Rusling
James F
JF
D002853:Chromatography, Liquid / Q000379:methods* / Q000592:standards; D004247:DNA / Q000187:drug effects; D018736:DNA Adducts / Q000032:analysis*; D004249:DNA Damage; D004563:Electrochemistry / Q000379:methods; D004852:Epoxy Compounds / Q000633:toxicity; D006147:Guanine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008741:Methyl Methanesulfonate / Q000633:toxicity; D009152:Mutagenicity Tests / Q000295:instrumentation* / Q000379:methods; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards
D018736:DNA Adducts; D004852:Epoxy Compounds; D006147:Guanine; D004247:DNA; C013690:styrene oxide; D008741:Methyl Methanesulfonate