Flow-Injection Assay of Superoxide Dismutase Based on the Reduction of Highly Water-Soluble Tetrazolium.

DOI PDF Web Site Web Site Web Site 被引用文献12件 参考文献18件
  • UKEDA Hiroyuki
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • SARKER Ashok K.
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • KAWANA Daisuke
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • SAWAMURA Masayoshi
    Department of Bioresources Science, Faculty of Agriculture, Kochi University

この論文をさがす

抄録

A flow-injection analysis (FIA) for superoxide dismutase (SOD) activity was developed based on the use of tetrazolium salt, WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate sodium salt) and an enzyme reactor packed with Sepharose 4B on which xanthine oxidase (XO) and catalase were co-immobilized. WST-1 is highly water-soluble, and no adhesion of the reduced form to the FIA line was observed during continuous operation for 3 months. As optimized conditions, a sample (9 vol) was mixed with a reagent solution (1 vol) containing 3 mM hypoxanthine and 2 mM WST-1, and the mixed solution (20 μl) was injected into a carrier stream of 50 mM carbonate buffer (pH 10.2) at a flow rate of 0.4 ml/min. Under the conditions, the concentration of the SOD preparation giving 50% inhibition (IC50) was 2.7 μg/ml and the sampling frequency was 30 samples/h. When the SOD activity in erythrocytes of rats was determined by the present FIA method, the values were linearly related to those obtained by the conventional nitroblue tetrazolium (NBT) assay (r=0.975; n=18). The enzyme reactor was stable for at least 200 repetitive injections.

収録刊行物

  • Analytical Sciences

    Analytical Sciences 15 (4), 353-357, 1999

    社団法人 日本分析化学会

被引用文献 (12)*注記

もっと見る

参考文献 (18)*注記

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ