A THERMOSPRAY LIQUID-CHROMATOGRAPHY MASS-SPECTROMETRY METHOD FOR ANALYSIS OF HUMAN URINE FOR THE MAJOR MALONDIALDEHYDE GUANINE ADDUCT

A method is described for detection and quantitation of the major malondialdehyde-guanine adduct (M1G) based on thermospray liquid chromatography/mass spectrometry. A stable isotope analog of M1G ([H-2(2)]M1G) was used as an internal standard. Thermospray mass spectra of M1G and [H-2(2)]M1G showed intense protonated molecular (MH+) ions that were suitable for use in quantitation of M1G. M1G was purified from human urine and reduced with NaBH4 to a dihydro derivative that was cleanly separated from the contaminants in the urine. The detection limit of reduced M1G by thermospray liquid chromatography/mass spectrometry in the selected ion monitoring mode was 250 fmol on column. Six human urine samples were analyzed, and the concentrations of M1G were below the limit of detection of the assay (500 fmol/mL).