固定化ピルビン酸オキシダーゼおよび固定化乳酸オキシダーゼを用いた血漿中ピルビン酸,乳酸の自動蛍光測定法

Abstract

A highly sensitive method for the automated fluorometric determination of pyruvate and lactate in human plasma using immobilized pyruvate oxidase and immobilized lactate oxidase in column form that does not require a blank correction is described. The method was based on the fluorometric determination of hydrogen peroxide formed by the action of pyruvate oxidase or lactate oxidase. Since the addition of sodium chloride to the recipient solution improved the dialysis rate of pyruvate by approximately three times, this method did not require a blank correction for the constituents that occurred in the fluorescent reaction. The present method was found to give perfect linearity of the data up to 0.4 mM for pyruvate and 8.0 mM for lactate with satisfactory precision, reproducibility and accurate reaction recoveries. The immobilized enzyme reactor unit showed good operational stability for a 5-week period, during which time it was repeatedly used for analysis over 2000 times. The results correlated satisfactorily with those obtained by well-established chemical and enzymatic methods.