Analysis of the Malondialdehyde−2‘- Deoxyguanosine Adduct Pyrimidopurinone in Human Leukocyte DNA by Gas Chromatography/Electron Capture/Negative Chemical Ionization/Mass Spectrometry

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Abstract
A method is described for the assay of the major malondialdehyde−deoxyguanosine adduct (M1G) based on immunoaffinity purification and gas chromatography/electron capture/negative chemical ionization/mass spectrometry. A stable isotope of M1G-deoxyribose ([2H2]M1G-dR) was used as an internal standard. Recovery of internal standard throughout the entire assay procedure was ∼40%. The assay showed a linear response over a range of 10−1000 pg of M1G-dR and was verified by analysis of a synthetic, M1G-containing oligomer. The limit of detection in biological samples was 100 fmol/sample, corresponding to 3 adducts/108 bases for 1 mg of DNA. DNA was isolated from the blood of 10 healthy human donors, and M1G levels were measured. A mean value of 6.2 ± 1.2 adducts/108 bases was obtained, with no obvious differences based on age or cigarette smoking. A small, but statistically significant difference was observed between the levels in females (5.1 ± 0.4 adducts/108 bases) and males (6.7 ± 1.1 adducts/108 bases). The presence of M1G in leukocyte DNA was further verified by analysis using liquid chromatography/electrospray ionization mass spectrometry.

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