Abstract
High-performance liquid chromatographic (HPLC) systems are described for the separation of the cardioactive metabolites of the digoxin and digitoxin series that are formed by the splitting of digitoxose sugar residues from the aglycone steroid: (1) isocratic separation of digoxin and its cardioactive metabolites; (2) isocratic separation of digitoxin and its cardioactive metabolites; and (3) gradient elution separation of the digoxin and digitoxin series including β-acetyl- and β-methyldigoxin. Separations were performed on a 10-μm bonded octadecyl phase column using various mixtures of acetonitrile—water and acetonitrile—methanol—water as the mobile phase. These methods provide high peak resolution and are well suited for collecting elution fractions, e.g. to link up with sensitive immunological measurements. An HPLC—radioimmunoassay method is described for the quantitation of digoxin, digitoxin and their metabolites in human tissues.
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