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Development of a high-performance liquid chromatographic-post-column fluorogenic assay for digoxin in serum
1Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, Canada.
Journal of Chromatography
|November 24, 1989
Summary
A new high-performance liquid chromatography assay offers sensitive and specific quantification of digoxin. This method effectively separates digoxin from metabolites for accurate clinical measurements.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Pharmacology
Background:
- Accurate quantification of digoxin is crucial for therapeutic drug monitoring.
- Existing assays may lack specificity or sensitivity for digoxin and its metabolites.
- Development of a robust analytical method is needed for clinical laboratories.
Purpose of the Study:
- To develop and validate a quantitative, sensitive, and specific assay for digoxin.
- To enable accurate measurement of digoxin in biological samples like serum.
- To differentiate digoxin from its known metabolites.
Main Methods:
- High-performance liquid chromatography (HPLC) coupled with post-column (PC) fluorogenic derivatization.
- Utilized a specific octadecylsilyl HPLC column and a defined mobile phase composition.
- Employed concentrated hydrochloric acid for PC derivatization and monitored fluorescence detection.
Main Results:
- The developed HPLC-PC assay demonstrated high specificity for digoxin, distinguishing it from metabolites and steroids.
- Linear detector response for digoxin was observed in the range of 0.5 to 3.3 ng.
- An effective serum extraction procedure was established to minimize interference from endogenous compounds.
Conclusions:
- The developed HPLC-PC assay is a quantitative, sensitive, and specific method for digoxin determination.
- This assay is suitable for measuring digoxin in serum, overcoming interference issues.
- The method provides a reliable tool for therapeutic drug monitoring of digoxin.