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Ultrafiltration in serum protein binding determinations

J B Whitlam, K F Brown

    Journal of Pharmaceutical Sciences
    |February 1, 1981
    PubMed
    Summary
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    Ultrafiltration accurately measures drug concentrations in serum, even for potent drugs with low free fractions. This technique is reliable as it does not disturb binding equilibria during filtration.

    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Pharmacology

    Background:

    • Accurate determination of serum protein binding is crucial for understanding drug efficacy and toxicity.
    • The ultrafiltration technique is widely used for clinical serum binding determinations.
    • Previous studies noted unexplained lower ultrafiltrate concentrations at higher filtration pressures.

    Purpose of the Study:

    • To theoretically and experimentally evaluate the ultrafiltration technique for clinical serum binding determinations.
    • To investigate the effect of filtration pressure on ultrafiltrate concentration.
    • To determine if ultrafiltration disturbs binding equilibria.

    Main Methods:

    • Theoretical evaluation using free energy considerations.
    • Mathematical simulations of molecular separation during ultrafiltration.

    Related Experiment Videos

  • Experimental verification using salicylate, ibuprofen, and carprofen in various matrices (buffer, purified proteins, whole serum).

    • Main Results:

    • Ultrafiltrate concentration approaches true free concentration as filtration pressure approaches zero.
    • A theory was developed to explain lower ultrafiltrate concentrations at higher pressures.
    • Mathematical simulations showed constant ultrafiltrate concentration during filtration, indicating undisturbed binding equilibria.
    • Experimental results validated the theoretical findings across different drug concentrations and matrices.

    Conclusions:

    • The ultrafiltration technique is a valid method for clinical serum binding determinations.
    • Binding equilibria are not disturbed by the ultrafiltration procedure, removing the need for arbitrary volume restrictions.
    • This finding enhances the utility of ultrafiltration for analyzing strongly bound drugs with very low free fractions.