Development and Validation of an Analytical Method for Nitrosamines Detection via LC-MS/MS in Pharmaceutical APIs and Finished Products.

Nitrosamines, a group of chemical compounds that form through the reaction between primary or secondary amines and nitrosating agents, have raised serious concerns within the pharmaceutical industry due to their well-documented carcinogenic properties. Since their unexpected detection in several medications worldwide in 2018, regulatory agencies have intensified scrutiny over their presence in pharmaceutical products. These impurities can arise at various stages of drug synthesis, manufacturing, and storage, making their prevention and detection a critical aspect of pharmaceutical safety. Given the potential health risks, strict regulatory limits have been established, necessitating the use of highly sensitive analytical techniques capable of identifying nitrosamines at ultra-trace levels, often in the parts-per-billion (ppb) range. This master’s thesis presents the development and validation of novel analytical methods utilizing liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) to detect and quantify nitrosamines in both active pharmaceutical ingredients (APIs) and finished drug products. The methods were carefully optimized to enhance sensitivity and selectivity, enabling the detection of these impurities at sub-ppb concentrations. Beyond detection, this study also investigates the nitrosation process, shedding light on the conditions under which these harmful compounds form. Understanding these formation mechanisms is essential for assessing risks in pharmaceutical manufacturing and implementing effective preventive strategies. The validated methods demonstrated strong specificity, accuracy, and precision across diverse pharmaceutical matrices. By integrating targeted LC-MS/MS analysis with a comprehensive validation process, this research provides a reliable and robust approach for detecting nitrosamines at levels compliant with regulatory standards. As a result, these methods offer a valuable tool for ensuring pharmaceutical quality control and safeguarding public health.