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Related Experiment Videos

Corticosteroid analysis in biological fluids by high-performance liquid chromatography.

J Q Rose, W J Jusko

    Journal of Chromatography
    |March 1, 1979
    PubMed
    Summary

    A new high-performance liquid chromatography method accurately measures prednisone, prednisolone, and cortisol in biological samples. This assay is sensitive, specific, and reproducible, aiding pharmacokinetic studies.

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    Area of Science:

    • Pharmacology
    • Analytical Chemistry
    • Biochemistry

    Background:

    • Accurate quantification of corticosteroids in biological fluids is crucial for pharmacokinetic and clinical studies.
    • Existing methods may lack the required sensitivity, specificity, or reproducibility for simultaneous analysis.

    Purpose of the Study:

    • To develop and validate a sensitive, specific, and reproducible high-performance liquid chromatographic (HPLC) assay for the simultaneous determination of prednisone, prednisolone, and cortisol.
    • To utilize dexamethasone as an internal standard for enhanced accuracy.

    Main Methods:

    • Development of an HPLC assay using a microparticulate silica gel column.
    • Sample preparation involved methylene chloride extraction, followed by sodium hydroxide and water washes.

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  • Detection was performed using UV detection at 254 nm.

    • Main Results:

    • The assay demonstrated high sensitivity, with a detection limit greater than 15 ng for all four steroids (prednisone, prednisolone, cortisol, and dexamethasone).
    • Specificity was confirmed through dual-wavelength UV detection and radioimmunoassay.
    • The assay was successfully applied to pharmacokinetic studies, providing typical plasma concentration-time profiles.

    Conclusions:

    • A robust and reliable HPLC method for simultaneous corticosteroid analysis in biological fluids has been established.
    • This assay is suitable for pharmacokinetic investigations and clinical monitoring of prednisone and its metabolites.
    • The developed method offers improved sensitivity and specificity compared to previous analytical techniques.