DEVELOPMENT OF A METHOD FOR THE DETERMINATION OF PROPAFENONE IN BIOLOGICAL FLUIDS
Abstract
The aim of this study was to develop a method for determining propafenone in biological fluids (blood, urine) using high-performance liquid chromatography (HPLC) for therapeutic drug monitoring. The studies were carried out on model blood and urine samples, and the method was also tested on the blood of volunteers who had taken a therapeutic dose of propafenone. Isolation of the drug from urine was performed by chloroform extraction (three times, 5 mL each) after acidifying the sample with 0.2 N hydrochloric acid solution to pH 3.5–4.0 and adding sodium chloride. Isolation from blood was performed using an acetate buffer solution (pH 4.0) followed by centrifugation and threefold extraction with chloroform. After evaporation of the organic solvent, the dry residue was dissolved in the mobile phase and chromatographed by HPLC. The analysis was carried out on an Agilent 1100 Series chromatograph equipped with a UV detector (detection at 242 nm), using a Spherisorb SCN column (4.6Ч120 mm, 3 µm). The mobile phase was a mixture of acetonitrile and acetate buffer solution at pH 6.0 (50:50), with a flow rate of 0.7 mL/min. HPLC analysis showed that the retention time of propafenone isolated from blood samples was 3.4 min. Co-extracted substances did not interfere with the determination of the drug. The quantitative content of propafenone was calculated by comparing the peak areas of the standard and test solutions. For the analysis of blood from volunteers who had taken a therapeutic dose of propafenone (150 mg), 5 mL blood samples were collected. Isolation of the drug was carried out according to the developed method, and propafenone was determined by HPLC under the conditions described above. The method allowed the drug to be quantified over the a range 0.04–0.06 mg/mL.
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