PMID: 2498017May 1, 1989Paper

Concurrent measurement of flecainide acetate and propranolol by normal-phase high-performance liquid chromatography

Clinical Chemistry
C L Rognerud, C N Ou

Abstract

This simple, isocratic, normal-phase liquid-chromatographic method concurrently measures flecainide acetate and propranolol in 100 microL of serum within 8 min. The chromatographic system consists of a Waters "Resolve" column packed with 5-microns silica spheres and a mobile phase of ammonium sulfate (10 mmol/L, pH 6.8)/methanol (22/78 by vol), pumped at 0.9 mL/min and monitored by a fluorometer (excitation at 225 nm and emission at 340 nm). After 100 microL of serum is mixed with 200 microL of the internal standard solution [N-(2-piperidylmethyl)-2,3-bis(2,2,2-trifluoroethoxy)-benzamide HCl, 2500 micrograms/L] and 200 microL of 0.2 mol/L sodium carbonate, the sample is extracted into butanol/hexane (20/80 by vol). The organic layer is separated and evaporated, and the residue is redissolved in 200 microL of methanol; 50 microL of this is injected onto the column. Relative recovery was 100% over the assay range of 25-2000 micrograms/L for flecainide and 10-2000 micrograms/L for propranolol. Within-run CVs were less than 2% for flecainide and less than 5% for propranolol; day-to-day CVs ranged from 5.0% to 6.5% for flecainide and from 3% to 12% for propranolol.

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