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High-performance liquid chromatography with electrochemical detection for determination of the major malondialdehyde-guanine adduct.

Goda Y, Marnett LJ
Chem Res Toxicol. 1991 4 (5): 520-4

PMID: 1793799 · DOI:10.1021/tx00023a005

A method is described for quantitative analysis of the pyrimidopurinone adduct (M1G) arising from reaction of malondialdehyde (MDA) with DNA. DNA samples treated with MDA are reduced with sodium borohydride and hydrolyzed with 0.1 N HCl. The bases released are isolated by solid-phase extraction and analyzed by high-performance liquid chromatography with electrochemical detection. 5,6-Dihydro-M1G (H2M1G) is detected with a glassy carbon electrode at an applied potential of 700 mV vs Ag/AgCl. At this potential, interference from normal deoxynucleoside bases is low. The limit of detection of H2M1G eluted from an HPLC column is 100-200 fmol. The method described should be useful for quantitation of M1G in a variety of DNA samples and biological fluids.

MeSH Terms (10)

Borohydrides Chromatography, High Pressure Liquid DNA Electrochemistry Guanine Hydrochloric Acid Hydrogen-Ion Concentration Hydrolysis Malondialdehyde Oxidation-Reduction

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