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Related Experiment Videos

A fluorimetric assay for cortisol.

Daniel Appel1, Rolf D Schmid, Calin-Aurel Dragan

  • 1Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.

Analytical and Bioanalytical Chemistry
|September 15, 2005
PubMed
Summary

This study presents a simple, rapid, and sensitive fluorimetric assay for quantifying cortisol. The method uses a novel fluorescent dye formation and is suitable for screening microorganisms involved in steroid hydroxylation.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry

Background:

  • Cortisol is a vital steroid hormone.
  • Accurate quantification of cortisol is essential for various biological and medical applications.
  • Existing methods for cortisol determination may lack simplicity, speed, or sensitivity.

Purpose of the Study:

  • To develop a simple, rapid, and sensitive fluorimetric assay for cortisol determination.
  • To establish a method suitable for preliminary screening of microorganisms capable of steroid hydroxylation.

Main Methods:

  • Cortisol is incubated with a mixture of sulfuric acid and acetic acid to form a fluorescent dye.
  • 2-methyl-4-pentanone is used as a solvent for extraction and fluorescence amplification.
  • Fluorescence is measured at specific excitation and emission wavelengths (max extinction at 475 nm, max emission at 525 nm).

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  • Assay validation involved biotransformation using recombinant Schizosaccharomyces pombe.

    • Main Results:

    • The assay achieves a limit of detection of 2.7 muM, eliminating the need for solvent evaporation.
    • Minor interference was observed from 11-deoxycortisol.
    • Typical standard deviations were below 4%, indicating good precision.
    • The assay was successfully validated using a biotransformation process.

    Conclusions:

    • The developed fluorimetric assay is simple, rapid, sensitive, and precise for cortisol quantification.
    • The method is suitable for preliminary screening of microorganisms for steroid hydroxylation capabilities.
    • This assay offers a valuable tool for research in endocrinology and biotechnology.