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Development of a high-performance liquid chromatographic assay for digoxin using post-column fluorogenic

E Kwong, K M McErlane

    Journal of Chromatography
    |April 25, 1986
    PubMed
    Summary

    A new high-performance liquid chromatography (HPLC) method accurately quantifies digoxin in plasma. This method uses a fluorogenic post-column reactor for sensitive detection of digoxin and its metabolites.

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    Area of Science:

    • Analytical Chemistry
    • Pharmacology

    Background:

    • Digoxin is a critical medication requiring precise therapeutic drug monitoring.
    • Existing methods for digoxin quantification may lack sensitivity or efficiency.

    Purpose of the Study:

    • To develop and optimize an efficient high-performance liquid chromatography (HPLC) method for digoxin and metabolite quantification in plasma.
    • To establish a sensitive and reliable assay for determining plasma digoxin levels.

    Main Methods:

    • Separation of digoxin and metabolites using a reversed-phase C18 HPLC column.
    • Post-column derivatization employing ascorbic acid, hydrogen peroxide, and hydrochloric acid to induce fluorescence.
    • Detection using a fluorescence detector after a heated reaction coil.

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    Main Results:

    • Achieved linear detector responses for digoxin from 1.5 to 10 ng on-column.
    • Demonstrated high sensitivity, linearity, and stability suitable for plasma digoxin determination.
    • Total analysis time, including post-column derivatization, was approximately 40 minutes.

    Conclusions:

    • The developed HPLC method provides an efficient and sensitive means for quantifying digoxin in plasma.
    • This assay is well-suited for routine therapeutic drug monitoring of digoxin.