固定化クレアチニナーゼを利用した血清クレアチニンのけい光測定法の研究

Abstract

A fluorimetric method using immobilized creatininase is described for the determination of creatinine in serum by continuous-flow analysis. In this method, creatinine in serum was dialyzed against acceptor buffer solution with a dialyzer after conversion of preformed creatine with creatine kinase-pyruvate kinase coupled enzymatic system and transformed to creatine by the immobilized creatininase column. The creatine reacted with ninhydrin in strongly alkaline solution to form highly fluorescent products, which were measured fluorimetrically. This reaction has been used as the basis for a simple, sensitive fluorimetric method for the determination of creatine in serum and urine. The present method offers the advantages of automation, high sensitivity, high precision, small sample volume and enhanced specificity owing to the use of enzyme. In addition, the method gave a linear standard curve for creatinine concentration up to 100 mg/l and the results well correlated with those obtained by Jaffe's method.