Assay of plasma semicarbazide-sensitive amine oxidase and determination of its endogenous substrate methylamine by liquid chromatography.

摘要: Abstract Semicarbazide-sensitive amine oxidase (SSAO) is present in plasma, as well other tissues. Previous studies indicated that SSAO of important physiological and pathophysiological functions. HPLC methods were developed for the assay determination plasma methylamine, an SSAO's endogenous substrate. Benzylamine was used artificial substrate enzyme activity SSAO. A 0.2-ml aliquot incubated with benzylamine at 37 °C 30 min. Benzaldehyde, enzymatic reaction product, derivatized 2,4-dinitrophenylhydrazine (DNPH), analyzed UV detection. defined benzaldehyde (nmol) formed per ml hour. Recoveries spiked to between 63.5 68.2% relative standard deviation less than 3%. To determine methylamine a 0.1-ml deproteinized by trichloroacetic acid centrifugation. The supernatant dansyl chloride fluorescence ppb (ng/ml) level 93.7 97.6% 2.5%.