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Related Experiment Videos

An enzymatic method for 5-fluorocytosine.

C M Huang1, M H Kroll, M Ruddel

  • 1Department of Clinical Pathology, NIAID, Bethesda, MD 20892.

Clinical Chemistry
|January 1, 1988
PubMed
Summary

A new enzymatic method accurately measures 5-fluorocytosine (5-FC) in serum using creatine iminohydrolase. This cost-effective assay is suitable for routine or emergency clinical use.

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Area of Science:

  • Clinical Chemistry
  • Enzymology
  • Analytical Chemistry

Background:

  • 5-fluorocytosine (5-FC) is an antifungal agent requiring therapeutic drug monitoring.
  • Accurate and efficient methods for 5-FC determination in serum are crucial for patient management.

Purpose of the Study:

  • To develop and validate a novel enzymatic assay for quantifying 5-fluorocytosine in human serum.
  • To assess the accuracy, precision, and potential interferences of the developed method.

Main Methods:

  • Enzymatic determination of 5-FC using creatine iminohydrolase (EC 3.5.4.21) on a Cobas-Bio analyzer.
  • Quantification of ammonia produced during the enzymatic reaction.
  • Validation using analytical recovery studies and comparison with High-Performance Liquid Chromatography (HPLC).

Main Results:

  • Excellent analytical recovery (y = 0.97x - 0.7, r = 0.997) over the 6.25-150 mg/L range.
  • High agreement with HPLC (y = 0.98x + 1.34), demonstrating method comparability.
  • Good precision with coefficients of variation (CVs) <3% (within-run) and <6% (between-run).
  • Identified interferences from bilirubin and lipemia (triglycerides >5 g/L); no interference from ammonia, glucose, urea, or hemolysis.

Conclusions:

  • The developed enzymatic assay provides an accurate, inexpensive, and user-friendly method for 5-FC serum determination.
  • The assay is robust and adaptable for both routine and emergency clinical laboratory settings.
  • Careful consideration of potential interferences from bilirubin and high triglyceride levels is necessary.

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