Determination of Possible Potential Genotoxic Impurities in Lenalidomide Drug Substance by Simple RP-HPLC Method

Abstract

This study is concerned with development and validation of HPLC method for the simultaneous detection and quantification of methyl 2-(chloromethyl)-3-nitrobenzoate (MCN), methyl 2-(bromomethyl)-5-nitrobenzoate (MMM), methyl 2-(bromomethyl)-6-nitrobenzoate (MON), methyl 2-(bromomethyl)-4-nitrobenzoate (MPN) and 2-methyl-3-nitrobenzoic acid methyl ester (MNM), which are the genotoxic impurities of lenalidomide. Chromatographic separation was accomplished using a Waters HPLC system equipped with Ascentis Express F5 (150 × 4.6 mm, 2.7 μm) using mobile phase composed of solvent A (0.1% perchloric acid): solvent B (methanol 80% and acetonitrile 20%); 55:45, vol/vol. The selected impurities were detected using UV detector set at 210 nm. The standard curves showed linearity in the range of concentrations 4.59-91.2 ppm (for MCN), 6.58-90.0 ppm (for MMM), 3.96-89.1 ppm (for MON), 6.47-89.7 ppm (for MPN) and 4.28-90.1 ppm (for MNM). The statistical results of method precision, system precision, specificity, accuracy, ruggedness was found to be within limits of acceptance. All the impurities were stable in lenalidomide test samples up to 24 h.