Immobilized Enzyme-Based Microtiter Plate Assay for Ethanol in Alcoholic Beverages

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A method for the determination of ethanol using alcohol oxidase (AOD), peroxidase (POD), and 2,2′-azino-bis(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS) was developed in which those enzymes were separately immobilized on the inside surface of each well of a 96-well microtiter plate. Prior to immobilization, the surface was activated with glutaraldehyde. The activation conditions remarkably affected the activity of the immobilized AOD. A sample solution containing ethanol was first added into the well with immobilized AOD (AOD-well). After 30 min, an aliquot (20 µl) was transferred into the well with immobilized POD containing 180 µl of 2 mM ABTS and the absorbance change at 415 nm for 10 min was measured with a microplate reader. The calibration curve was linear over the range 0.10 - 1.0 mM, and the relative standard deviation (n=5) was 0.8% for 1 mM ethanol. The ethanol concentration in alcoholic beverages, obtained by the present method, agreed well with that by the F-kit method. The AOD-well was reusable for 20 successive determinations.

収録刊行物

  • Analytical sciences : the international journal of the Japan Society for Analytical Chemistry  

    Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 15(5), 447-450, 1999-05 

    社団法人 日本分析化学会

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各種コード

  • NII論文ID(NAID)
    10002419704
  • NII書誌ID(NCID)
    AA10500785
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    09106340
  • NDL 記事登録ID
    4721773
  • NDL 雑誌分類
    ZP4(科学技術--化学・化学工業--分析化学)
  • NDL 請求記号
    Z54-F482
  • データ提供元
    CJP書誌  NDL  J-STAGE 
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