A Spectrophotometric Assay of Histaminase Activity based on the Hydrazone Derivative of Imidazole Acetaldehyde with 2,4-Dinitrophenylhydrazine

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The activity of histaminase by using histamine as a substrate was spectrophotometrically estimated as the hydrazone derivative of imidazole acetaldehyde with 2,4-dinitrophenylhydrazine (DNP). The measurement of enzyme activity in pig kidney was based on oxygen consumption in the presence of histamine by means of oxygen electrode, and the assay based on hydrazone derivative was performed with a spectrophotometer. In determination of oxygen consumption, there was a remarkable difference in the patterns of activities under the conditions of incubation temperature of 38°and 60°. The maximum activity of histaminase at 60°was about 2.4 times higher than that at 38°when the concentration of histamine was 10^<-4>M. By measuring the absorbance of the hydrazone derivative of imidazole acetaldehyde formed in the presence of 3×10^<-4>M histamine at 60°, it could be compared with oxygen consumption proportional to an increased amount of enzyme. Isooctane extractable hydrazones and unreacted DNP disturbed markedly the spectrophotometric assay, but these substances could be removed by use of a mixed solvent of isooctane and CHCl_3. If aldehyde compound is formed over 1 μM/min or if oxygen is consumed over 15μM/min even in the incubation temperature of 38°, it is indicated that hydrazone derivative of aldehyde compound can be spectrophotometrically detected and that the assay method can be applied to the measurement of histaminase activity.
公益社団法人日本薬学会の論文
1977-01-25

A Spectrophotometric Assay of Histaminase Activity based on the Hydrazone Derivative of Imidazole Acetaldehyde with 2,4-Dinitrophenylhydrazine

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