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Related Experiment Videos

pH lability in serum during equilibrium dialysis.

O Brørs, S Jacobsen

    British Journal of Clinical Pharmacology
    |July 1, 1985
    PubMed
    Summary

    Serum pH changes during dialysis can cause significant errors in drug binding measurements. Using buffers that maintain a stable pH is crucial for accurate unbound drug fraction determination.

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    Area of Science:

    • Biochemistry
    • Pharmacokinetics
    • Analytical Chemistry

    Background:

    • Accurate determination of unbound drug fractions in serum is critical for therapeutic drug monitoring and understanding drug efficacy.
    • Equilibrium dialysis is a common method for measuring drug binding to serum proteins.
    • Variations in serum pH can significantly impact drug-protein interactions and thus the unbound fraction.

    Purpose of the Study:

    • To investigate the pH changes in human serum during equilibrium dialysis.
    • To evaluate the influence of different buffers and conditions on serum pH stability.
    • To highlight the implications of pH variability on the accuracy of unbound drug fraction measurements.

    Main Methods:

    • Human serum (native or pH-adjusted) was dialyzed against various buffers (sodium phosphate, Krebs Ringer phosphate, Krebs Ringer bicarbonate) at different temperatures (22°C, 37°C) and buffer molarities.
    • Serum pH was monitored over a 3-hour period during dialysis.
    • Changes in pH were correlated with dialysis conditions.

    Main Results:

    • Native serum (initial pH 7.7-7.9) exhibited pH fluctuations during dialysis, decreasing initially then increasing.
    • pH-adjusted serum showed a continuous increase in pH throughout dialysis.
    • Increased temperature, lower buffer molarity, and native serum conditions exacerbated pH changes.
    • These pH shifts were substantial enough to introduce significant errors in unbound drug fraction calculations.

    Conclusions:

    • Equilibrium dialysis of human serum can lead to significant pH variations, particularly under certain conditions.
    • These pH changes directly impact the accuracy of unbound drug fraction measurements.
    • It is essential to utilize buffers that ensure a stable and appropriate pH level during equilibrium dialysis for reliable drug binding studies.

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