A novel assay for determination of sulfated bile acids in urine by use of flow-injection chemiluminescence principle with immobilized enzymes

✍ Scribed by Xiu-Feng Gao; Kazunoli Ikebukuro; Isao Karube; Yong-Sheng Li

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 256 KB
• Volume: 9
• Category: Article
• ISSN: 0895-7533
• DOI: 10.1002/(sici)1098-2728(1997)9:2<69::aid-lra4>3.0.co;2-s

✦ Synopsis

Multistep enzymatic reactions take place when sulfated bile acids (SBAs) pass through an immobilized enzyme reactor. First, SBAs take place in the reaction of desulfation under catalytical action of a bile salt sulfatase immobilized in the reactor and formed 3b-hydroxyl bile acids. Formed 3b-hydroxyl bile acids react with nicotinamide adenine dinucleotide (b-NAD‫)ם‬ under catalysis of another enzyme of 3b-hydroxysteroid dehydrogenase coimmobilized in the column and are converted to 3-ketosteroid. At the same time, b-NAD‫ם‬ is changed into reduced nicotinamide adenine dinucleotide (NADH). Looking as if chain reaction, 1-MPMS taken as an electron mediator reacts immediately with NADH coexisting in the carrier solution and is turned into 1-MPMSH 2 .

Formed 1-MPMSH 2 again reacts with dissolved oxygen existing in the carrier solution and produces hydrogen peroxide. Last, the hydrogen peroxide reacts with the luminol reagent and gives out light in the