Issue 6, 2008

Sensitive determination of a glyoxal–DNA adduct biomarker candidate by column switching capillary liquid chromatographyelectrospray ionization mass spectrometry

Abstract

A method based on column switching packed capillary liquid chromatography electrospray mass spectrometry has been developed for the determination of the adduct glyoxaldeoxyguanosine, a biomarker candidate for the assessment of glyoxal exposure, in DNA hydrolysate solutions. Microgram amounts of DNA were isolated and enzymatically hydrolyzed to deoxyribonucleosides, prior to ultrafiltration and subsequent dilution to a sample solution consisting of wateracetonitrileformic acid (98 : 2 : 0.2, v/v). The sample solution was loaded onto a 1 mm I.D. × 5 mm Hypercarb (5 μm) porous graphitic carbon trap column for analyte enrichment using an injection volume of 200 μl, and was subsequently back-flushed onto a 0.30 mm I.D. × 150 mm Lichrospher diol (5 μm) analytical column. The samples were loaded with a flow rate of 40 μl min−1 and glyoxaldeoxyguanosine was desorbed from the trap column and eluted with an isocratic mobile phase consisting of wateracetonitrileformic acid (50 : 50 : 0.2, v/v) at a flow rate of 5 μl min−1. Mass spectrometric determination of glyoxaldeoxyguanosine was obtained by multiple reaction monitoring of the transition [M + H]+m/z 326 → m/z 210. The method was evaluated over the concentration range 0.25–50 ng ml−1 of glyoxaldeoxyguanosine in the hydrolysate of 5 μg DNA. The method was linear with a correlation coefficient of 0.9998 in this range. The within-day (n = 6) and between-day (n = 6) precisions were determined as 1.2–11% and 1.4–11% RSD, respectively, and the recovery was close to 100%. The mass limit of detection was 15 pg, corresponding to a concentration limit of detection of 75 fg μl−1 DNA hydrolysate solution, corresponding to 48 adducts per 106 normal nucleosides. The method was applied for the determination of glyoxaldeoxyguanosine in DNA hydrolysate solutions of calf thymus DNA and cell cultures after reaction or incubation with glyoxal.

Graphical abstract: Sensitive determination of a glyoxal–DNA adduct biomarker candidate by column switching capillary liquid chromatography electrospray ionization mass spectrometry

Article information

Article type
Paper
Submitted
04 Jan 2008
Accepted
03 Mar 2008
First published
26 Mar 2008

Analyst, 2008,133, 802-809

Sensitive determination of a glyoxalDNA adduct biomarker candidate by column switching capillary liquid chromatography electrospray ionization mass spectrometry

R. Olsen, S. Øvrebø, S. Thorud, E. Lundanes, Y. Thomassen, T. Greibrokk and P. Molander, Analyst, 2008, 133, 802 DOI: 10.1039/B719842F

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