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Related Experiment Videos

Fluorometric determination of urea by flow injection analysis.

M S Abdel-Latif1, G G Guilbault

  • 1Department of Chemistry, University of New Orleans, Louisiana 70148.

Journal of Biotechnology
|April 1, 1990
PubMed
Summary

This study presents a fluorometric method for urea determination using immobilized urease and O-phthalaldehyde (OPA). While effective, serum analysis requires a blank correction due to interfering fluorescent compounds.

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Area of Science:

  • Analytical Chemistry
  • Biochemistry

Background:

  • Urea analysis is crucial in clinical diagnostics.
  • Existing methods may lack selectivity or require complex sample preparation.

Purpose of the Study:

  • To develop a sensitive and selective fluorometric method for urea determination.
  • To immobilize urease for enhanced stability and reusability in flow injection analysis.

Main Methods:

  • Urea determination via fluorometry utilizing flow injection analysis.
  • Enzymatic reaction of ammonia (from urea hydrolysis) with O-phthalaldehyde (OPA) in an alkaline medium.
  • Immobilization of urease onto a Pall Immunodyne membrane via covalent bonding.

Main Results:

  • The OPA method demonstrated high fluorescence selectivity for ammonia.

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  • Immobilized urease provided a stable enzyme preparation for the enzymatic reactor.
  • Direct analysis of urea in serum was complicated by endogenous fluorescent compounds, necessitating a blank correction.

    • Conclusions:

    • The developed fluorometric flow injection analysis method offers a sensitive approach for urea quantification.
    • Enzyme immobilization enhances the stability and practicality of the analytical system.
    • Serum urea analysis using this method requires a blank run to account for background fluorescence and ensure accurate results.