| 摘要 |
PURPOSE:To perform the analysis of bile acid while keeping immobilized enzyme stable for a long period of time, by a method wherein NAD<+> is premixed in an elution liquid for separating a bile acid-containing specimen and an NaCl solution is added to the eluate from a separation column while the resulting solution mixture is passed through a specific immobilized enzyme column to detect formed NADH. CONSTITUTION:A definite amount of an elution liquid having a reactive reagent NAD<+> mixed therein is supplied by a pump 22 and a bile acid-containing specimen is injected from a sampler 23 to be flowed to a chromatograph separation of the elution liquid 21. The eluate 31 and a 0.1-0.5M NaCl solution 25 sent by a pump 26 are mixed in a mixer 27 while the resulting solution mixture is passed through a column 28 packed with 3-hydroxysteroid hydrogenase (3alpha-HSD) immobilized by a solid carrier to form NADH which is, in turn, detected by a spectrophotometer 29 of which the excitation light is set to 350nm and the generated fluorescence to 460nm and the peak of each bile acid component in the specimen or the content thereof is displayed or recorded by a recorder 30. |
