Determination of endogenous and exogenously derived N7-(2-hydroxyethyl)guanine adducts in ethylene oxide-treated rats.
Ethylene oxide (EO) is one of the most widely used intermediates in the chemical industry. It is also formed endogenously as a result of cytochrome P450-mediated metabolism of ethylene, which is ubiquitous in the environment. Additionally, ethylene is generated in vivo during normal physiological processes such as methionine oxidation and lipid peroxidation; therefore, humans are continually exposed to EO. EO is classed by the IARC as carcinogenic to humans and reacts with DNA, primarily forming N7-(2-hydroxyethyl)guanine adducts (N7-HEG), which can be used as biomarkers of exposure and potential cancer risk. To assess the risks to humans associated with occupational exposure to low EO concentrations, it is necessary to establish the relative contribution of DNA damage arising from endogenous and exogenously derived EO. Using a newly developed highly sensitive LC-MS/MS assay with selected reaction monitoring that offers a limit of detection of 0.1 fmol of N7-HEG on column, we have established background levels of N7-HEG (1.1-3.5 adducts/10(8) nucleotides) in tissues of rats. Following intraperitoneal administration of a single dose or three daily doses of EO (0.01-1.0 mg/kg), N7-HEG adducts generally increased with dose, except at the lowest concentration where total N7-HEG levels were no different to that detected in control animals, indicating that any increase was negligible as compared to the endogenous damage already present. In the 3 day study, the kinetics of adduct removal were also investigated and in comparing N7-HEG formation in the two studies, DNA damage did not appear to accumulate with repeated administration.
Marsden
Debbie A
DA
Cancer Biomarkers and Prevention Group, Department of Cancer Studies and Molecular Medicine, The Biocentre, University of Leicester, Leicester, LE1 7RH, United Kingdom.
Jones
Donald J L
DJ
Lamb
John H
JH
Tompkins
Elaine M
EM
Farmer
Peter B
PB
Brown
Karen
K
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D018736:DNA Adducts / Q000032:analysis* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005027:Ethylene Oxide / Q000008:administration & dosage* / Q000493:pharmacokinetics; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis; D007274:Injections, Intraperitoneal; D008297:Male; D015394:Molecular Structure; D051381:Rats; D011916:Rats, Inbred F344; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods; D013997:Time Factors; D014018:Tissue Distribution
D018736:DNA Adducts; C035018:N(7)-hydroxyethylguanine; D006147:Guanine; D005027:Ethylene Oxide