Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR DETERMINATION OF STRUCTURE RELATED GENOTOXIC IMPURITIES IN OMEPRAZOLE DRUG BY LC-MS/MS

*S. Sirish Chandra, G. Nagasowjanya, A. Ajitha and Dr. V. Uma Maheswara Rao

ABSTRACT

A simple, sensitive and accurate gradient reversed-phase LC-MS/MS method has been developed for the Determination of structure related impurities namely 2-chloro methyl -3,5-dimethyl-4-methoxy pyridine Hcl (CDMP), 4-methoxy-3,5-dimethyl-4-nitro pyridine-1-oxide (DNO) and 4-methoxy-2-amino aniline (MAA) in Omeprazole (API). These are Potential genotoxic impurities and hence need to be controlled in Omeprazole (API) before formulating. The analysis was Performed using SHIMADZU LC-MS/MS (tandem mass spectrometer) 8030 C18 column (YMC PACK PRO, 100×4.6, 3 μm) as a stationary phase with Column oven temperature ambient and PDA detection at 254nm. The separation was achieved using mobile phase Buffer (0.1%formic acid) pH 4: Methanol. The method was optimized based on the peak shapes and by comparing Retention time between 2-chloro methyl -3, 5-dimethyl-4- methoxy pyridine Hcl (CDMP), 4-methoxy-3, 5-dimethyl-4-nitro pyridine-1-oxide (DNO), 4-methoxy-2-amino aniline (MAA) and Omeprazole. The method was validated as per International Conference of Harmonization (ICH) guidelines in terms of linearity, precision, accuracy and specificity. The LOD and LOQ values were found to be 1.8μg/ml and 3.1μg/ml, respectively. The sample Concentration injected was 1mg/ml. The method is linear within the range of 3.1-18.6μg/ml for the Impurities.

Keywords: Genotoxic impurities, Omeprazole, CDMP, DNO, MAA.