Simultaneous Determination of Procainamide and N-acetylprocainamide in Rat Plasma by Ultra-High-Pressure Liquid Chromatography Coupled with a Diode Array Detector and Its Application to a Pharmacokinetic Study in Rats

Abstract

A simple, sensitive, and reliable reversed-phase, Ultra-High-Pressure Liquid Chromatography (UHPLC) coupled with a Diode Array Detector (DAD) method for the simultaneous determination of Procainamide (PA) and its major metabolite, N-acetylprocainamide (NAPA), in rat plasma was developed and validated. A simple deproteinization method with methanol was applied to the rat plasma samples, which were analyzed using UHPLC equipped with DAD at 280 nm, and a Synergi™ 4 µm polar, reversed-phase column using 1% acetic acid (pH 5.5) and methanol (76:24, v/v) as eluent in isocratic mode at a flow rate 0.2 mL/min. The method showed good linearity (r2 > 0.998) over the concentration range of 20–100,000 and 20–10,000 ng/mL for PA and NAPA, respectively. Intra- and inter-day accuracies ranged from 97.7 to 110.9%, and precision was <10.5% for PA and 99.7 to 109.2 and <10.5%, respectively, for NAPA. The lower limit of quantification was 20 ng/mL for both compounds. This is the first report of the UHPLC-DAD bioanalytical method for simultaneous measurement of PA and NAPA. The most obvious advantage of this method over previously reported HPLC methods is that it requires small sample and injection volumes, with a straightforward, one-step sample preparation. It overcomes the limitations of previous methods, which use large sample volume and complex sample preparation. The devised method was successfully applied to the quantification of PA and NAPA after an intravenous bolus administration of 10 mg/kg procainamide hydrochloride to rats.

Physiologically-Based Pharmacokinetic Modeling for Drug-Drug Interactions of Procainamide and N-Acetylprocainamide with Cimetidine, an Inhibitor of rOCT2 and rMATE1, in Rats

“…The concentrations of PA and NAPA in various matrices (e.g., plasma, urinary samples, and samples from in vitro studies) were determined as previously described [36]. Briefly, analyses were performed with an Agilent Technologies 1290 Infinity II UHPLC system (Agilent Technologies, Palo Alto, CA, USA) equipped with a multisampler (G7167B), a flexible pump (G7104A), a multicolumn thermostat (MCT) (G7116B), and a DAD detector (G7117A).…”

A Rapid UPLC-MS Method for Quantification of Gomisin D in Rat Plasma and Its Application to a Pharmacokinetic and Bioavailability Study

“…ESI positive and negative modes were both compared and evaluated in method development [24,25,26]. In this work, the response of the positive ion mode showed a better signal-to-noise ratio than that of the negative ion mode.…”

Metabolic Stability of D-Allulose in Biorelevant Media and Hepatocytes: Comparison with Fructose and Erythritol

“…Experimental animal protocols were reviewed and approved by the Animal Care and Use Committee of the Gachon University (#GIACUC-R2018022, approval date 7 November 2018). As described in our recent pharmacokinetic study [26], overnight fasted rats were anesthetized intramuscularly with Zoletil (20 mg/kg), and a femoral vein and artery were then surgically cannulated with polyethylene tubing (PE50; Clay Adams, Parsippany, NJ, USA). The cannula was flushed with 20 IU/mL heparinized saline to prevent clotting.…”