Abstract
A procedure is described for the quantification of the major malondialdehyde deoxyguanosine adduct, pyrimido[1,2-α]purin-10(3 H)-one-deoxyribose (M 1GdR) in urine. M 1GdR is isolated from urine by a combination of C 18 solid-phase extraction and immunoaffinity chromatography. Sodium borohydride treatment reduces M 1GdR to the 5,6-dihydro derivative, which is quantified by liquid chromatography–mass spectrometry. Authentic [7,9 - 15 N,8 - 13 C] M 1 GdR is added to urine as an internal standard. A detection limit of 50 fmol M 1GdR/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 1GdR are below the detection limit of the assay. This method is easily adaptable to the analysis of M 1GdR in DNA samples or biological fluids.
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