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

DOI NDLデジタルコレクション Web Site Web Site 被引用文献2件 参考文献21件

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タイトル別名
  • Determination for sulfated bile acids using an immobilized enzyme reactor based on FIA chemiluminescence.
  • 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

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抄録

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.

収録刊行物

  • 分析化学

    分析化学 46 (5), 357-365, 1997

    公益社団法人 日本分析化学会

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