Precolumn Fluorescence Derivatization of Carboxylic Acids Using 4-Aminomethyl-6, 7-dimethoxycoumarin in a Two-Phase Medium

• Published in: Analytical Sciences Vol. 12; no. 2; pp. 189 - 193
• Main Authors: SASAMOT, Kazumi, USHIJIMA, Tamano, SAITO, Mikihiko, OHKURA, Yosuke
• Format: Journal Article
• Language: English
• Published: Tokyo The Japan Society for Analytical Chemistry 1996; Japan Society for Analytical Chemistry; Japan Science and Technology Agency
• Subjects: Biological and medical sciences; carboxylic acid; coumarin; Derivatization; Errors of metabolism; fluorescence detection; high-performance liquid chromatography; Lipids (lysosomal enzyme disorders, storage diseases); Medical sciences; Metabolic diseases; Performance evaluation; Human; Biological fluid; Fluorescence detector; Coumarine derivatives; HPLC chromatography; Lipids; Metabolic diseases; Derivatization; Fatty acids; Analysis method; Carboxylic acid; Clinical biology; Serum; Quantitative analysis
• ISSN: 0910-6340; 1348-2246
• DOI: 10.2116/analsci.12.189

A new coumarin-based amine, 4-aminomethyl-6, 7-dimethoxycoumarin, was synthesized as a precolumn fluorescence derivatization reagent for carboxylic acids. The derivatization using the amine in a two-phase medium of water and ethyl acetate was examined. The two-phase derivatization method with the amine was found to be of value since it allows simple and rapid HPLC assays of carboxylic acids with a reasonable sensitivity and reparability. The detection limit was determined to be 20-50fmol per 10μl injection volume at a signal-to-noise ratio of 3 under partially optimized conditions. The method was also applied to the assay of fatty acids in human serum.