<原報>Ninhydrin-Phenylacetaldehyde による第一アミン類のけい光分析
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概要
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McCaman and Robins introduced a fluorometric method for assay of serum Phenylalanine which is based on interaction with ninhydrin and peptides. Based on this observation, Samejima et al. developed a fluorometric determination of primary amines, amino acids, peptides and aminosugars by using phenylacetaldehyde and ninhydrin. The structure of fluorophor produced in the reaction has been elucidated by M. Weigele et al. who explained the mechanism of the reaction. In the course of structure study, a new reaction was discovered which led to the synthesis of fluorescamine as a reagent for determination of primary amines. Although fluorogenic reaction by fluorescamine proceeds at room temperature in weakly alkaline medium, the reagent has also some drawbacks, for instance, low solubility in water and ease of degradation in the presence of water which some times result in operational difficulties. In the course of our study to improve the water solubility of fluorescamine, we found little investigation has been done to estimate the original method of Samejima et al. as an analytical procedure for primary amino groups, especially in comparison with fluorescamine. As a consequence, we have examined the usefulness of Samejima's procedure and found the original procedure has a few advantages over fluorescamine procedure. In the present investigation, we have examined the conditions required for ninhydrin fluorescent procedure and modified the reaction condition for the micro assay by combining High Pressure Liquid Chromatograph and also developed the system for automated determination of primary amines at picomols level.
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