Validated liquid chromatographic method for the determination of N-3-(2,2,5,5-tetramethyl-3-pirrolin-3-carboxamidopropylphthalimide hydrochloride), a novel antiarrhythmic agent in human plasma.

A simple high-performance liquid chromatographic method with ultraviolet absorbance detection has been developed to determine the concentration of N-3-(2,2,5,5-tetramethyl-3-pirrolin-3-carboxamidopropylphthalim ide hydrochloride; A-2545), a new antiarrhythmic agent from human plasma. Separation of the investigated compound and internal standard was achieved on a Nucleosil 7 C18 column with a 0.01-M potassium dihydrogenphosphate buffer (pH 2.5)-methanol (60:40, v/v) mobile phase. The detection was performed at 220 nm. During the determinations, buspirone served as the internal standard. The compounds were isolated from plasma on a Bakerbond C18 solid-phase extraction cartridge and the mean absolute recovery was 92.9%. The limit of quantitation was found to be 10 ng/ml. The bioanalytical method was validated with respect to linearity, within- and between-day accuracy and precision, system suitability and stability. All validated parameters were found to be within the internationally required limits. The developed analytical method for A-2545 was found to be suitable for application in pharmacokinetic studies and for human drug monitoring.