ペルオキシダーゼ固定電極の臨床検査への応用

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In clinical examinations many biochemical substances are monitored by measuring H_2O_2 (HP) or NAD(P)H photometrically with the use of suitable oxidoreductases. A peroxidase (PO)-entrapped and ferrocence (FC)-embedded carbon paste electrode with a dialysis membrane (cut-off molecular weight of 100) on the surface of the PO layer (mPOFCE) allowed HP determination sensitive to levels as low as 50nM in constant-potential amperometry in the potential ranges from 0 to 0.2V at pH ranges from 5.0 to 9.5. In the photometric analysis, proton donors interfere with the color reaction of pigment with HP catalyzed by PO. A proton donor such as NADH can not be in contact with PO by the use of mPOFCE. NADH produced in a assay mixture can be detected with mPOFCE in the presence of 5-methyl phenazinemethosulfate (MPMS), where MPMS catalyzes NAD(P)H oxidation with dioxygen to generate HP. By the use of this electrode, many clinically important substances in human serum could be monitored easily by adding auxiliary enzymes and some reagents in the test solution based on the measurement of reductive current due to FC+ produced by the reduction of HP. 1: Creatine, 2: creatinine, 3: cholesterol, 4: HDLcholesterol, 5: oxalate, 6: urate and 7: glucose were determined by the use of creatinase (1), creatininase (1, 2), sarcosine oxidase (1, 2), cholesterol oxidase and esterase (3), polyethylene glycol-modified cholesterol esterase and oxidase (4), oxalate oxidase (5), uricase (6) and glucose oxidase (7). In the presence of NAD(P) and MPMS, 1: lactate dehydrogenase, 2: isocitrrate dehydrogenase, 3: leucine aminopeptidase (using leucine dehydrogenase), and 4: alanine aminotransferase and aspartate aminotransferase (using glutamate dehydrogenase, GTD) were assayed. In the presence of NADPH, MPMS and GTD, ammonia was determined.
活水女子大学の論文

ペルオキシダーゼ固定電極の臨床検査への応用

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