Determination for sulfated bile acids using an immobilized enzyme reactor based on FIA chemiluminescence.

DOI NDL Digital Collections Web Site Web Site 2 Citations 21 References Open Access
  • GAO Xiu-Feng
    Division of Medical Sciences, University of Tokyo
  • JI Hong Seok
    Research Center for Advanced Science and Technology, University of Tokyo
  • IKEBUKURO Kazunori
    Research Center for Advanced Science and Technology, University of Tokyo
  • BABA Shigeaki
    Hyogo Medical Center for Adults
  • KARUBE Isao
    Research Center for Advanced Science and Technology, University of Tokyo
  • LI Yong-Sheng
    Laboratory of Analytical Chemistry, Department of Applied Chemistry, Northeast China Institute of Electrical Power Engineering

Bibliographic Information

Other Title
  • フローインジェクション分析／固定化酵素化学発光法による硫酸抱合胆汁酸の迅速定量法
  • フロー インジェクション ブンセキ コテイカ コウソ カガク ハッコウホウ ニ

Search this article

Description

An accurate, simple, less time-consuming and sensitive chemiluminescence(CL)-FIA method has been established for the determination of sulfated bile acids (SBA) based on two kinds of immobilized enzymes. After SBA is desulfated under catalysis of a bile acid sulphate sulfatase immobilized in an enzymatic reactor, it is changed into 3β-hydroxyl bile acids. The formed 3β-hydroxyl bile acid reacts upon nicotinamide adenine dinucleotide(β-NAD+) under another catalyst of 3β-hydroxysteroid dehydrogenase coimmobilized, and is changed into 3-ketosteroid; at same time β-NAD+ is turned to nicotinamide adenine dinucleotide (NADH). NADH reduces molecular oxygen in the carrier to O2- and hydrogen peroxide (H2O2) in the presence of l-MPMS as an electronic mediator. Last, the produced O2- and H2O2 react with the luminol reagent from another tubing line, and give out light in the presence of POD. Consequently, SBA can be determined by the CL intensity. Based on the principle mentioned above, and our FIA manifold as well as the experimental results optimized in the paper, a kind of new-type analytical instrument for clinic tests of SBA in urine or blood will be developed. The sampling frequency of the method is 30 sample/h, and its relative standard deviation is less than 2.2%. The calibration curve of the method also shows good linearity over a range of 0.112 μM glycolithocholic acid 3-sulphate as a standard.

Journal

  • BUNSEKI KAGAKU

    BUNSEKI KAGAKU 46 (5), 357-365, 1997

    The Japan Society for Analytical Chemistry

Citations (2)*help

See more

References(21)*help

See more

Keywords

Details 詳細情報について

Report a problem

Back to top