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Kinetic enzymatic method for determining serum creatinine.

G A Moss, R J Bondar, D M Buzzelli

    Clinical Chemistry
    |September 1, 1975
    PubMed
    Summary
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    This study introduces a new enzymatic method for measuring serum creatinine. The assay is rapid, accurate, and unaffected by common interfering substances, offering an improvement over the traditional Jaffé test.

    Area of Science:

    • Clinical Chemistry
    • Biochemical Assays
    • Enzymology

    Background:

    • Accurate measurement of serum creatinine is crucial for assessing kidney function.
    • The direct Jaffé method, while widely used, suffers from interferences and requires deproteinization.
    • There is a need for a more specific and efficient creatinine assay.

    Purpose of the Study:

    • To develop and validate a novel enzymatic method for serum creatinine determination.
    • To compare the performance of the new method with the direct Jaffé test.

    Main Methods:

    • Utilized creatinine amidohydrolase for the initial hydrolysis of creatinine.
    • Employed a coupled enzymatic reaction involving creatine kinase, pyruvate kinase, and lactate dehydrogenase.
    • Monitored the change in absorbance at 340 nm to quantify creatinine concentration.

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  • Assessed the effect of common anticoagulants and metabolites on assay accuracy.

    • Main Results:

    • The enzymatic method demonstrated linear correlation between absorbance and creatinine concentration up to 100 mg/liter.
    • Analytical recovery of added creatinine averaged 102% in both normal and abnormal sera.
    • The new method showed significantly lower results compared to the direct Jaffé test, indicating higher specificity.
    • The assay is rapid, completing within approximately 10 minutes, and requires only 250 µL of serum.
    • Common substances like heparin, oxalate, and glucose did not interfere at concentrations up to 1.0 g/liter.

    Conclusions:

    • The described enzymatic procedure provides a rapid, specific, and reliable method for serum creatinine measurement.
    • This assay overcomes the limitations of the Jaffé method, offering improved accuracy and reduced interference.
    • The method is suitable for routine clinical laboratory use due to its efficiency and minimal sample requirements.