Suzuki M, Yoshida M
Clin Chim Acta. 1984 Nov 15;143(2):147-55. doi: 10.1016/0009-8981(84)90222-5.
We describe a new colorimetric determination of serum creatinine which does not require a blank to correct for endogenous creatine. In the first reaction, creatinase (creatinase amidinohydrolase EC 3.5.3.3) and sarcosine oxidase (sarcosine: oxygen oxidoreductase (demethylating) EC 1.5.3.1) were used in the enzymatic hydrolysis of endogenous creatine to produce hydrogen peroxide. In the presence of horseradish peroxidase, 2,4-dichlorophenolsulfonate (2,4-DCPS) was converted to a colorless polymer by hydrogen peroxide. In the second reaction, creatininase (creatinine amidohydrolase EC 3.5.2.10) and 4-aminoantipyrine (4-AA) were added, and only the creatine generated from creatinine by creatininase was hydrolyzed sequentially by creatinase and sarcosine oxidase to produce hydrogen peroxide. This newly-formed hydrogen peroxide was measured at 510 nm in a coupled reaction catalyzed by peroxidase, with 2,4-DCPS/4-AA as a chromogen. The standard curve was linear up to 20 mg/dl; 40 microliter of serum and 20 min were required for determination. Analytical recovery of creatinine added to either normal or abnormal sera averaged 98.5%. Within-day and day-to-day studies gave CV values of less than or equal to 2.9% and less than or equal to 4.8%, respectively. No significant interferences were observed with the proposed method. The results obtained by the present method correlated well with those obtained by the Jaffé procedure.
我们描述了一种新的血清肌酐比色测定法，该方法无需空白对照来校正内源性肌酸。在第一个反应中，肌酸酶（肌酸酶脒基水解酶，EC 3.5.3.3）和肌氨酸氧化酶（肌氨酸：氧氧化还原酶（去甲基化），EC 1.5.3.1）用于内源性肌酸的酶促水解以产生过氧化氢。在辣根过氧化物酶存在下，2,4 - 二氯苯酚磺酸盐（2,4 - DCPS）被过氧化氢转化为无色聚合物。在第二个反应中，加入肌酐酶（肌酐酰胺水解酶，EC 3.5.2.10）和4 - 氨基安替比林（4 - AA），只有肌酐酶从肌酐产生的肌酸依次被肌酸酶和肌氨酸氧化酶水解以产生过氧化氢。在过氧化物酶催化的偶联反应中，以2,4 - DCPS / 4 - AA作为显色剂，在510 nm处测量新形成的过氧化氢。标准曲线在高达20 mg/dl时呈线性；测定需要40微升血清和20分钟。添加到正常或异常血清中的肌酐的分析回收率平均为98.5%。日内和日间研究的CV值分别小于或等于2.9%和小于或等于4.8%。所提出的方法未观察到明显干扰。本方法获得的结果与Jaffé法获得的结果相关性良好。
