Nitrosamine Impurities

The presence of nitrosamine impurities (NNAs) in pharmaceutical products has become a great challenge for the pharmaceutical industry. Since the so-called “Nitrosamine (N-NAs) Crisis” emerged in June 2018, the European Medicines Agency (EMA), and the Food and Drug Administration (FDA), along with the rest of the regulatory agencies, began to carry out Risk Analysis of Nitrosamines, in order to evaluate the different causes of formation of these compounds and eradicate their presence, both in active ingredient and in finished product. Therefore, mitigating contamination by children and adolescents is one of the global regulatory priorities for the pharmaceutical industry. In order to respond to many of the needs that have arisen, the Center for Applied Chemistry and Biotechnology (CQAB), within the Fine Chemicals and Biotechnology Pilot Plant Area, has focused on carrying out the synthesis, isolation and characterization of NNAs and N-Nitroso compounds (NOCs) that can be formed directly on the corresponding active ingredients (APIs) and/or on medications. Said compounds are carried out in the presence of nitrosating agents and/or under oxidative alkylation conditions. To date, NNAs have been obtained in active ingredients, synthetic intermediates and reagents with great molecular diversity and application in different therapeutic areas. These nitrosamines have emerged after carrying out a detailed risk analysis of the API synthesis route, considering both the intermediates and reagents used, as well as the corresponding formulations in the finished product. NNAs are synthesized and used as standards when quantified by NMR. Thus, our clients can address compliance with ICH Q3.

Furthermore, we also offer analytical support associated with risk analysis. In the Bioanalysis and Quality Control Area, we are actively working on the development of chromatographic methods for the detection and quantification of different nitrosamines at very low levels in various products. When this problem arose, a new ultrasensitive analytical method based on the FDA method was developed and validated using LC-MS/MS with HRMS for the quantification of six nitrosamine (NA) impurities initially set by the Agencies in different drugs ( N-nitroso-dimethylamine (NDMA), N-nitroso-diethylamine (NDEA), N-nitroso-ethyl-isopropylamine (NEIPA), N-nitroso-diisopropylamine (NDIPA), N-nitroso-dibutylamine (NDBA) and N-acid nitroso-methyl-4-aminobutyric (NMBA)In addition, this method improves the sensitivity of the FDA method up to about 20 times, also reducing the analysis time by half. Its high sensitivity and easy sample treatment should be of relevance in the field of pharmaceutical quality control for the determination of NA impurities associated with the synthesis processes. In addition, for the detection of other possible nitrosamines formed as byproducts due to the modification of manufacturing processes, a screening method is also developed in Full scan showing detection limits similar to those of the FDA. Based on the Maximum Daily Dose of each drug, the quantification and detection levels of the corresponding nitrosamines are established, developing a specific analytical method for the determination of 10% of the Limit Test of each one.


Therefore, thanks to the synergy between the services offered by the two Areas, the CQAB provides itself with the appropriate tools to provide a reliable and effective solution, to guarantee correct detection, identification and quantification of these mutagenic impurities in the chemical-pharmaceutical industry.