Fluorometric assay of amino acids and amines by use of 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole(NBD-F) in high-performance liquid chromatography |
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Authors: | K. Imai Y. Watanaba T. Toyo'oka |
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Affiliation: | (1) Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113 Tokyo, Japan |
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Abstract: | Summary The reaction of a newly developed fluoregenic reagent, 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole(NBD-F), with amino acids and biogenic amines was investigated. NBD-F was reactive to both primary and secondary amines including amino acids and biogenic amines such as catecholamines. The amino acids were reacted with the reagent, separated by high-performance liquid chromatography on -Bondapak C18 and detected at 10 to 100 fmol level. A few g of protein hydrolysates, rabbit pyruvate kinase M1, rabbit aldolase A and papain, were adequate for the amino acids quantitation. An automatic amino acid analyzer with fluorometric detection by the post-column derivatization with NBD-F enabled the amino acid profile analysis in blood samples present in a paper disc of 3 mm diameter.Presented at the 14th International Symposium on Chromatography London, September, 1982 |
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Keywords: | Liquid chromatography Derivatization with NBD-F Amino acids, biogenic amines Blood analysis |
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