4-ブロモメチル-7-メトキシクマリンにより螢光標識化された5-フルオロウラシルおよびピリミジンヌクレオシド類の高速液体クロマトグラフィーへの応用

概要

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A method for the labeling of 5-fluorouracil (5-FU) and four pyrimidine nucleosides (uridine, deoxyuridine, fluorodeoxyuridine and thymidine) with 4-bromomethyl-7-methoxycoumarin (Br-Mmc) using 18-crown-6 as a catalyst is described. The fluorescent derivatives are prepared by adding 5.0mg of Br-Mmc, 1.0mg of 18-crown-6 and 25 mg of potassium carbonate to a solution containing 0.5mg of pyrimidine compounds in 20 ml acetone. The mixture is refluxed for 30min at 75℃. The excess Br-Mmc is treated with n-valeric acid after completion of the reaction. High performance liquid chromatographic separation of these fluorescence derivatives have been obtained on a 25cm column packed with octadecylsilica. The amount of the derivatives is linearly related to the amount of the starting 5-FU and pyrimidine compounds and the procedure can therefore be used in the quantitative analysis. The procedure is simple and requires little or no experience in derivatization techniques. The detection limits of 5-FU, uridine, deoxyuridine, fluorodeoxyuridine and thymidine are 2.5 pg, 5.0 pg, 50 pg, 50 pg and 75pg, respectively. The structures of the fluorescence derivatives are also discussed from the results of thin layer chromatography.
社団法人日本薬学会の論文
1983-09-25