4-ニトロフェニル-β-グルクロニドを基質にした透析膜を被覆した炭素電極を用いる血清中β-グルクロニダーゼ活性のアンペロメトリック測定(<特集>医療と分析化学)

概要

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The magnitude of the current increase for five min due to the oxidation of p-nitrophenol (PNP) released from p-nitrophenyl-β-glucuronide (PNPGD) by β-glucuronidase(βGlu) from a snail was proportional to the concentration of the enzyme ranging from 0.3 to 3Ul^<-1> . The steady state current by PNP was measured at 1 V in acetate buffer of pH 5 stirred at 500rpm using a dialysis membrane-covered glassy carbon electrode, and was proportional to the concentration of PNP from 1 to 500μM. Since the βGlu activities of serum were very small, they were determined by measuring the difference in current at 0 and 120min after a reaction of acetate buffer solution containing 10mM PNPGD and serum diluted by 1/5. The βGlu activities of twelve serum samples obtained by the present method and a colorimetric method are in good agreement with the correlation coefficient of 0.96 over the range 0.8〜3.6Ul^<-1>. This method, which is free from a process for color development, is simple compared with a colorimetric method. The K_M value of serum βglu for PNPGD was determined to be 2mM.
1997-12-05