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Improved ultrafiltration method for determining unbound cortisol in plasma

I Jerkunica, J Sophianopoulos, D Sgoutas

    Clinical Chemistry
    |November 1, 1980
    PubMed
    Summary
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    This study presents a new ultrafiltration technique for accurately measuring unbound cortisol in plasma. The method is fast, requires minimal sample volume, and offers reproducible results comparable to equilibrium dialysis.

    Area of Science:

    • Biochemistry
    • Endocrinology

    Background:

    • Accurate measurement of unbound cortisol is crucial for diagnosing adrenal disorders.
    • Existing methods like equilibrium dialysis can be time-consuming and require larger sample volumes.

    Purpose of the Study:

    • To develop and validate a modified ultrafiltration method for quantifying unbound cortisol in plasma or serum.
    • To assess the method's speed, practicality, and reproducibility.

    Main Methods:

    • A modified ultrafiltration technique using [3H]cortisol for equilibration.
    • Collection of two successive 0.2-mL protein-free ultrafiltrate fractions.
    • Analysis of the second fraction for unbound cortisol proportion.

    Main Results:

    • The assay is rapid, completing in under 2 hours.

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  • It requires a small sample volume (≤1.5 mL).
  • The method demonstrated high reproducibility (CV: 4.5% within-assay, 5.2% between-assays) and correlated well with equilibrium dialysis results across various patient groups.

    • Conclusions:

    • The modified ultrafiltration method provides a rapid, practical, and reproducible means of measuring unbound cortisol.
    • This technique is suitable for clinical applications, including diagnosing conditions like Cushing's disease and adrenal insufficiency.