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An improved ultrafiltration method for determining free testosterone in serum

I Vlahos, W MacMahon, D Sgoutas

    Clinical Chemistry
    |November 1, 1982
    PubMed
    Summary
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    This study introduces a rapid and reproducible method for measuring free testosterone in serum using centrifugal ultrafiltration. The assay accurately quantifies free testosterone, showing better resolution in hirsute women compared to total testosterone measurements.

    Area of Science:

    • Endocrinology
    • Clinical Chemistry
    • Biochemical Assays

    Background:

    • Accurate measurement of free testosterone is crucial for diagnosing conditions like hirsutism.
    • Existing methods for free testosterone determination can be time-consuming or require large sample volumes.

    Purpose of the Study:

    • To develop and validate a rapid, practicable, and reproducible assay for free testosterone in serum.
    • To compare the diagnostic utility of free versus total testosterone measurements in different populations.

    Main Methods:

    • Utilized Amicon MPS-1 centrifugal ultrafiltration device with YMB membrane.
    • Combined total testosterone assay with ultrafiltrable fraction of added [3H]testosterone.
    • Determined unbound fraction in ultrafiltrates from diluted serum equilibrated with [3H]testosterone.

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    Main Results:

    • Assay is rapid (<1 hour), practicable (0.6 mL serum), and reproducible (CV 3.2% intra-assay, 3.9% inter-assay).
    • Accuracy validated by comparing free testosterone in ultrafiltrate of dialyzed serum to prior dialysate.
    • Results agreed with equilibrium dialysis; free testosterone provided better discrimination in hirsute women than total testosterone.

    Conclusions:

    • Centrifugal ultrafiltration offers a valid and efficient method for free testosterone measurement.
    • Free testosterone assay demonstrates superior resolution for differentiating certain endocrine conditions, particularly in women.
    • This method enhances the clinical utility of testosterone measurements in endocrinology.