High-Performance Liquid Chromatographic Determination of 3-Methylcytosine in Deoxyribonucleic Acid Treated with Carcinogenic Methylating Agents in Vitro and in Vivo

概要

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A high-performance liquid chromatographic method is described for the determination of 3-methylcytosine (m^3Cyt) in deoxyribonucleic acid (DNA). Calf thymus DNA was methylated with N-methyl-N-nitrosourea or N-methyl-N'-nitro-N-nitrosoguanidine in vitro. Methylated rat liver DNA was obtained by administration of dimethylnitrosamine to rats. The DNA samples were hydrolyzed with perchloric acid at 100℃ for 1 h, and after the removal of perchloric acid by the use of anion exchange resin, the filtrate was concentrated and loaded on a Dowex 1×2 anion exchanger. The eluted cytosine derivatives were collected and injected into a liquid chromatograph. The content of m^3Cyt in methylated DNA was successfully determined by this method. The amount of m^3Cyt increased with the dosage of methylating agent in vitro, but showed no significant dose-dependent increase in vivo.
社団法人日本薬学会の論文
1985-03-25