title: Development of a method for determination of the malondialdehyde-deoxyguanosine adduct in urine using liquid chromatography-tandem mass spectrometry.
abstract: A procedure is described for the quantification of the major malondialdehyde deoxyguanosine adduct, pyrimido[1,2-alpha]purin-10(3H)-one-deoxyribose (M(1)GdR) in urine. M(1)GdR is isolated from urine by a combination of C(18) solid-phase extraction and immunoaffinity chromatography. Sodium borohydride treatment reduces M(1)GdR to the 5,6-dihydro derivative, which is quantified by liquid chromatography-mass spectrometry. Authentic [7,9-15N,8-13C]M(1)GdR is added to urine as an internal standard. A detection limit of 50 fmol M(1)GdR/ml urine is achieved starting with 5 ml of urine. Analysis of urine samples from control rats or rats treated with CCl(4) indicates that the levels of M(1)GdR are below the detection limit of the assay. This method is easily adaptable to the analysis of M(1)GdR in DNA samples or biological fluids.
authors.affiliation: Department of Biochemistry, Center in Molecular Toxicology, and Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt University, Nashville, TN 37232, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Otteneder
authors.forename: J Scott
authors.initials: JS
authors.lastname: Daniels
authors.forename: Markus
authors.initials: M
authors.lastname: Voehler
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Marnett
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D018736:DNA Adducts / Q000737:chemistry / Q000652:urine*; D003849:Deoxyguanosine / Q000737:chemistry*; D008297:Male; D008315:Malondialdehyde / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D015394:Molecular Structure; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D018736:DNA Adducts; D008315:Malondialdehyde; D003849:Deoxyguanosine