Fossati P, Prencipe L, Berti G
Clin Chem. 1983 Aug;29(8):1494-6.
We describe a new colorimetric method for measuring creatinine in serum and urine. Creatinine hydrolysis is catalyzed by creatinine amidohydrolase, and the creatine so produced is assayed in reactions catalyzed sequentially by creatine amidinohydrolase and sarcosine oxidase in a system that generates hydrogen peroxide. The hydrogen peroxide is measured at 510 nm in a reaction catalyzed by horseradish peroxidase, with 3,5-dichloro-2-hydroxybenzenesulfonic acid/4-aminophenazone as the chromogen. This series of reactions is complete in 30 min at room temperature. A blank sample measurement corrects for endogenous creatine. The standard curve is linear for creatinine concentrations as great as 2.21 mmol/L. Analytical recovery of creatinine in human sera and urine averaged 99.8%. Within-run and between-run precision studies gave CVs of less than or equal to 3.3 and less than or equal to 4.3% for a serum with a creatinine concentration of 69 mumol/L. Results by this method agree well (r greater than 0.99) with those by both the enzymic ultraviolet method of Wahlefeld and the fuller's earth/Jaffé method.
我们描述了一种用于测定血清和尿液中肌酐的新比色法。肌酐水解由肌酐酰胺水解酶催化，所产生的肌酸在一个产生过氧化氢的系统中，依次由肌酸脒基水解酶和肌氨酸氧化酶催化的反应中进行测定。在辣根过氧化物酶催化的反应中，以3,5 - 二氯 - 2 - 羟基苯磺酸/4 - 氨基苯甲酮作为显色剂，于510 nm处测定过氧化氢。这一系列反应在室温下30分钟内完成。空白样品测量可校正内源性肌酸。肌酐浓度高达2.21 mmol/L时，标准曲线呈线性。人血清和尿液中肌酐的分析回收率平均为99.8%。对于肌酐浓度为69 μmol/L的血清，批内和批间精密度研究的变异系数分别小于或等于3.3%和小于或等于4.3%。该方法的结果与瓦勒费尔德的酶促紫外法和硅藻土/贾菲法的结果高度一致（r大于0.99）。
