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Automated microanalysis of creatinine by coupled enzyme reactions
K Asano1, H Iwasaki, K Fujimura
1Central Research Laboratories, Kobayashi Pharmaceutical Co., Ltd., Osaka, Japan.
Hiroshima Journal of Medical Sciences
|March 1, 1992
Summary
This study presents an economical enzymatic assay for creatinine determination using sequential enzyme reactions. The optimized method is suitable for automated analyzers, requiring minimal sample volume for accurate results.
Area of Science:
- Clinical Chemistry
- Biochemistry
- Enzymology
Background:
- Creatinine is a key biomarker for kidney function.
- Accurate and efficient creatinine measurement is crucial for clinical diagnostics.
- Existing methods may require larger sample volumes or be less economical.
Purpose of the Study:
- To develop and optimize an enzymatic assay for creatinine determination.
- To adapt the assay for use with automated clinical analyzers.
- To establish an economical method for laboratories.
Main Methods:
- Sequential enzymatic reactions involving creatinine amidohydrolase, creatine amidinohydrolase, and sarcosine oxidase to generate hydrogen peroxide.
- Stoichiometric use of hydrogen peroxide for a colorimetric condensation reaction catalyzed by horse-radish peroxidase.
- Optimization of assay conditions for a HITACHI 7250 autoanalyzer.
Main Results:
- Successful generation of hydrogen peroxide from creatinine via a three-enzyme cascade.
- Formation of a quinone dye with maximum absorbance at 546 nm.
- Established optimized assay conditions for automated analysis.
Conclusions:
- The developed enzymatic assay provides an accurate method for creatinine determination.
- The assay is highly economical, requiring less than 5 microliters of sample.
- This method is well-suited for laboratories utilizing autoanalyzers for routine diagnostics.