Determination of Genotoxic Impurity (R)-(+)-1-Phenylethylamine in Pregabalin Drug Substances by High Performance Liquid Chromatography

Abstract

A highly sensitive high performance liquid chromatography (HPLC) method is developed and validated for the selective trace level determination of (R)-(+)-1-Phenylethylamine impurity, a potential genotoxic that may present in Pregabalin drug substances. HPLC column used was Prontosil 120-5-C18-ace-EPS (250 x 4.6 mm, 5µm). The gradient elution mode was selected where mobile phase-A consist of Buffer (40Mm Diammonium hydrogen o-phosphate solution adjusted to pH 6.5 with dilute orthophosphoric acid) and Methanol and Acetonitrile in ratio 8:1:1 while mobile phase-B as Acetonitrile, The flow rate was set to 0.8 mL/min and run time of 60 minutes. Selected wavelength 210 nm for the analysis on UV detector. The column oven temperature was kept at 25°C. The developed method was validated according to ICH guideline and found to be linear in the range of 1.31 ppm to 3.93 ppm with a correlation coefficient 0.9942. Limit of detection and limit of quantitation was found to be 0.29 ppm and 0.88 ppm respectively. Recovery for this impurity was found between 95.75% and 105.49%. Validation results prove that the method is specific, selective, precise and robust. The developed method can successfully be applied for the determination of genotoxic toxic impurity (R)-(+)-1-Phenylethylamine in Pregabalin.