Enantioselective analysis of sotalol in plasma by reversed-phase high-performance liquid chromatography using diastereomeric derivatives

Journal of Chromatography. B, Biomedical Applications
W D Hooper, P V Baker

Abstract

A procedure for the concurrent determination of the (+)- and (-)-enantiomers of sotalol in plasma using high-performance liquid chromatography of diastereomeric derivatives is described. Sotalol is extracted from a 0.5-ml aliquot of plasma at pH 9.3 using ethyl acetate. Atenolol is used as the internal standard. The ethyl acetate is removed under vacuum, and the residue derivatized with R-(-)-1-(1-naphthyl)ethyl isocyanate (NEIC, 0.005% in chloroform) in the presence of trace quantities of carbonate buffer. The chloroform is removed, the residue reconstituted in mobile phase (acetonitrile-water, 39:61, v/v), and an aliquot injected into the HPLC column. A C18 trapping column is used to retain excess derivatizing reagent. While the derivatives are separated on a C18 analytical column with the isocratic mobile phase mentioned above at 1.5 ml/min, the column-switching allows back-flushing of the trapping column to prepare for the next injection. The derivatives were detected using a fluorescence detector with excitation wavelength 280 nm and emission wavelength 320 nm. The method was fully validated, and shown to have excellent linearity, specificity, sensitivity, accuracy and precision. It has been applied to the determination of...Continue Reading

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