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J.Health Sci., 46(3), 219-222, 2000
Determination of Glucose in Plasma Using Immobilized Enzymes in a Flow System
Hisakazu Mori,*,a Miyuki Ando,a Yoshie Sugi,a Miyako Kimura,a and Katsuhiko Machidab
aKyoritsu College of Pharmacy, 1 -5 -30 Shibakoen, Minato-ku, Tokyo 105 -8512, Japan and bDepartment of Laboratory Medicine, Jikei University, School of Medicine, 3 -25 -8 Nishi-shinbashi, Minato-ku, Tokyo 105 -8461, Japan
Glucose was determined using an apparatus containing an enzyme reactor in a flow line. The enzymes used for glucose assay were hexokinase and glucose-6-phosphate dehydrogenase. NADH formed by enzymatic reactions was fluorometrically detected. The calibration curve for glucose was linear within the range of 0.2 -50 mu M (r=0.9996), and the detection limit was 0.1 mu M (S/N=3). This method was applied to the analysis of glucose in plasma. No influence of sodium fluoride on the activities of immobilized enzymes was observed. Recovery of glucose added to plasma or serum was found to be in the range of 98 to 106%. We investigated the correlation between the glucose content determined by other methods and that determined by the present method. A linear relationship was observed between that determined by the present method and that by both the glucose oxidase-electrode method (y=0.986x-4.6, r=0.9993) and the soluble enzyme method (y=1.004x+1.3, r=0.9982).
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