Fully Enzymatic Colorimetric Assay of Serum and Urine Creatinine Which Obviates the Need for Sample Blank Measurements
作者:
J. Siedel,
R. Deeg,
H. Seidel,
H. Möllering,
J. Staepels,
H. Gauhl,
J. Ziegenhorn,
期刊:
Analytical Letters
(Taylor Available online 1988)
卷期:
Volume 21,
issue 6
页码: 1009-1017
ISSN:0003-2719
年代: 1988
DOI:10.1080/00032718808071927
出版商: Taylor & Francis Group
关键词: Creatinine;serum and urine;enzymatic assay
数据来源: Taylor
摘要:
A fully enzymatic method for the colorimetric determination of serum and urine creatinine is described which does not require sample blank measurements. It is based on the formation of hydrogen peroxide from creatinine in a reaction sequence catalyzed by creatinine iminohydrolase, ATP-dependent 1-methylhydantoinase, N-carbamoylsarcosine amidohydrolase and sarcosine oxidase. The hydrogen peroxide is quantitated with high sensitivity at 546 nm by a chromogenic system consisting of peroxidase, 2′-sulpho-2-methyl-benzthiazolinone hydrazone and 2,4,6-tribromo-3-hydroxy-benzoic acid. Only 20 μL of sample are needed for the assay, the total reaction being complete within 10 min at 25°. Within-run precision gave a CV of 3.1 and 1.6 % at serum creatinine concentrations of 79 and 160 μmol/L, respectively, and the standard curve is linear up to at least 1760 μmol/L. The assay yields results which agree well with those found by both an enzymatic UV-method and an alternate enzymatic colorimetric procedure necesitating sample blank measurements to correct for endogenous creatine.
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