A simple, sensitive, and reliable reversed-phase, Ultra-High-Pressure Liquid\nChromatography (UHPLC) coupled with a Diode Array Detector (DAD) method for the simultaneous\ndetermination of Procainamide (PA) and its major metabolite, N-acetylprocainamide (NAPA), in rat\nplasma was developed and validated. A simple deproteinization method with methanol was applied\nto the rat plasma samples, which were analyzed using UHPLC equipped with DAD at 280 nm, and a\nSynergiââ??¢4 Ã?¼m polar, reversed-phase column using 1% acetic acid (pH 5.5) and methanol (76:24, v/v)\nas eluent in isocratic mode at a flow rate 0.2 mL/min. The method showed good linearity (r2 > 0.998)\nover the concentration range of 20ââ?¬â??100,000 and 20ââ?¬â??10,000 ng/mL for PA and NAPA, respectively.\nIntra- and inter-day accuracies ranged from 97.7 to 110.9%, and precision was <10.5% for PA and\n99.7 to 109.2 and <10.5%, respectively, for NAPA. The lower limit of quantification was 20 ng/mL for\nboth compounds. This is the first report of the UHPLC-DAD bioanalytical method for simultaneous\nmeasurement of PA and NAPA. The most obvious advantage of this method over previously reported\nHPLC methods is that it requires small sample and injection volumes, with a straightforward,\none-step sample preparation. It overcomes the limitations of previous methods, which use large\nsample volume and complex sample preparation. The devised method was successfully applied to the\nquantification of PA and NAPA after an intravenous bolus administration of 10 mg/kg procainamide\nhydrochloride to rats.
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