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Related Experiment Videos

Chlorthalidone analysis using carbonic anhydrase inhibition.

M M Johnston, H Li, D Mufson

    Journal of Pharmaceutical Sciences
    |December 1, 1977
    PubMed
    Summary

    This study developed an assay to measure chlorthalidone in urine. The method revealed rapid absorption and biphasic elimination of chlorthalidone in human subjects.

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    Area of Science:

    • Pharmacology
    • Analytical Chemistry
    • Biochemistry

    Background:

    • Chlorthalidone is a diuretic used to treat hypertension.
    • Accurate methods for quantifying chlorthalidone in biological samples are essential for pharmacokinetic studies.

    Purpose of the Study:

    • To develop and validate a sensitive assay for determining chlorthalidone concentrations in human urine.
    • To investigate the absorption and elimination kinetics of chlorthalidone following oral administration.

    Main Methods:

    • Enzyme inhibition assay using bovine erythrocyte carbonic anhydrase to measure chlorthalidone.
    • Spectrophotometric detection of p-nitrophenol formation at 400 nm.
    • Automated assay procedure for sample processing and data acquisition.

    Main Results:

    • The assay demonstrated a linear response for chlorthalidone in the range of 0.1-3.0 microgram/ml with a precision of +/- 0.05 microgram/ml.
    • No endogenous interfering substances were detected in urine samples from 26 healthy subjects.
    • Pharmacokinetic analysis of urine from six subjects after 100 mg oral chlorthalidone showed rapid absorption and biphasic elimination with half-lives of 1.5 hr (alpha-phase) and 35 hr (beta-phase).

    Conclusions:

    • An automated, sensitive, and specific assay for urinary chlorthalidone was successfully developed.
    • The study provides key pharmacokinetic data on chlorthalidone absorption and elimination in humans.
    • The biphasic elimination pattern suggests complex distribution and/or metabolism of chlorthalidone.

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