title: Development and application of an LC-MS/MS method for the detection of the vinyl chloride-induced DNA adduct N(2),3-ethenoguanine in tissues of adult and weanling rats following exposure to [(13)C(2)]-VC.
abstract: In the 1970s, exposure to vinyl chloride (VC) was shown to cause liver angiosarcoma in VC workers. We have developed a new LC-MS/MS method for analyzing the promutagenic DNA adduct N(2),3-ethenoguanine (εG) and have applied this to DNA from tissues of both adult and weanling rats exposed to 1100 ppm [(13)C(2)]-VC for 5 days or 1100 ppm VC for 1 day. This assay utilizes neutral thermal hydrolysis and an HPLC cleanup prior to quantitation by LC-MS/MS. The number of endogenous and exogenous εG adducts in DNA from tissues of adult rats exposed to [(13)C(2)]-VC for 5 days was 4.1 ± 2.8 adducts/10(8) guanine of endogenous and 19.0 ± 4.9 adducts/10(8) guanine of exogenous εG in the liver, 8.4 ± 2.8 adducts/10(8) guanine of endogenous and 7.4 ± 0.5 adducts/10(8) guanine of exogenous εG in the lung, and 5.9 ± 3.3 adducts/10(8) guanine of endogenous and 5.7 ± 2.1 adducts/10(8) guanine of exogenous εG in the kidney (n = 4). Additionally, the data from weanling rats demonstrated higher numbers of exogenous εG, with ∼4-fold higher amounts in the liver DNA of weanlings (75.9 ± 17.9 adducts/10(8) guanine) in comparison to adult rats and ∼2-fold higher amounts in the lung (15.8 ± 3.6 adducts/10(8) guanine) and kidney (12.9 ± 0.4 adducts/10(8) guanine) (n = 8). The use of stable isotope labeled VC permitted accurate estimates of the half-life of εG for the first time by comparing [(13)C(2)]-εG in adult rats with identically exposed animals euthanized 2, 4, or 8 weeks later. The half-life of εG was found to be 150 days in the liver and lung and 75 days in the kidney, suggesting little or no active repair of this promutagenic adduct.
authors.affiliation: Department of Environmental Sciences and Engineering, and Curriculum in Toxicology, Gillings School of Global Public Health, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Esra
authors.initials: E
authors.lastname: Mutlu
authors.forename: Leonard B
authors.initials: LB
authors.lastname: Collins
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Stout
authors.forename: Patricia B
authors.initials: PB
authors.lastname: Upton
authors.forename: Laura R
authors.initials: LR
authors.lastname: Daye
authors.forename: Darrell
authors.initials: D
authors.lastname: Winsett
authors.forename: Gary
authors.initials: G
authors.lastname: Hatch
authors.forename: Paul
authors.initials: P
authors.lastname: Evansky
authors.forename: James A
authors.initials: JA
authors.lastname: Swenberg
mesh_terms: D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry; D002273:Carcinogens / Q000737:chemistry* / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018736:DNA Adducts / Q000032:analysis*; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis / Q000493:pharmacokinetics; D006207:Half-Life; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014752:Vinyl Chloride / Q000737:chemistry* / Q000633:toxicity
chemical_list: D002247:Carbon Isotopes; D002273:Carcinogens; D018736:DNA Adducts; C015196:N(2),3-ethenoguanine; D006147:Guanine; D014752:Vinyl Chloride
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