High-performance liquid chromatographic method with electrochemical detection for the analysis of O6-methylguanine.
An improved system consisting of a combination of high-performance liquid chromatographic methods with electrochemical detection for the separation and analysis of the DNA adduct O6-methylguanine (O6MG) has been developed. This adduct is produced by the interaction of methylating agents with DNA and induces mispairing in the DNA of the target cells. A good separation of modified from unmodified bases is first achieved with an HPLC system using a Partisil 10 SCX column and a salt gradient. A second HPLC step with electrochemical detection and a C18 column is used for farther separation and quantitation of O6-methylguanine. This method shows a linear response up to 15 pg of 06MG tested. The lowest amount detected was 0.5 pg of O6MG and is highly reproducible. This method is useful to study DNA damage as a product of cellular metabolism and its effects on the process of carcinogenesis.
Vasquez
H
H
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston 77225, USA.
Seifert
W
W
Strobel
H
H
D000970:Antineoplastic Agents / Q000009:adverse effects; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003110:Colonic Neoplasms / Q000188:drug therapy; D004249:DNA Damage; D004563:Electrochemistry / Q000379:methods*; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis*; D006801:Humans; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D006147:Guanine; C008449:O-(6)-methylguanine