Advanced analytical techniques for detection, separation and quantification of N-nitroso and related impurities in trimetazidine: A UPLC-PDA, LC-MS/MS, and GC-MS study

Abstract Three techniques (UPLC, LC-MS/MS, and GC-MS) were developed to detect and evaluate various types of impurities in the trimetazidine API molecule. The UPLC method was developed to investigate simultaneously the nine process related impurities (IMP ‘A’ to ‘I’), GC-MS method was developed to quantify N-nitrosopiperazine and 1,4 dinitrosopiperazine simultaneously and LC-MS method for N-nitrosotrimetazidine quantification. Every studied impurity had good linear calibration curves over the concentration ranges of LOQ to 150% of the test sample concentration. The developed approaches exhibited the ability to quantify investigated impurities with great sensitivity, as seen by the LOQ values obtained, which were much below the impurity specification limits. The specificity, linearity, sensitivity, accuracy, precision, and robustness of the devised approaches were all validated in conformity with ICH guidance. There was also discussion on the potential mechanism for the N-nitrosotrimetazidine synthesis. This investigation showed that the established techniques (UPLC, LC-MS/MS, and GC-MS) for various forms of impurities in the trimetazidine API molecule may be utilised for quality control, as mandated by regulatory bodies to guarantee the product's safety in addition to effectiveness.