Since the so-called "Nitrosamines (N-NAs) Crisis" emerged in June 2018, the European
Medicines Agency (EMA), and the Food and Drug Administration (FDA), together with the
rest of the regulatory agencies, began to carry out risk analysis, in order to identify the
different causes of the formation of these compounds and eradicate their presence, both in
active ingredient and in finished product. And in order to respond to many of the needs that
have arisen from these risk analyses, the Center for Applied Chemistry and Biotechnology
(CQAB), within the Pilot Plant and Biotechnology Area, has focused on the synthesis,
isolation and characterization of N-NAs and N-Nitroso compounds (NOCs) formed directly
on the corresponding active ingredients (APIs). Until now, N-NAs have been obtained in
active ingredients with great molecular diversity and application in different therapeutic
areas. These API nitrosamines are being used as standards when quantified by NMR.
Currently, several of the structures obtained are being used to address a large part of the
problem of identifying N-NAs in APIs and finished products, thus favoring their correct
pharmacovigilance.
Moreover, a new ultra-sensitive analytical method based on the FDA method was developed using
LC-MS/MS with HRMS for the quantification of six nitrosamine (NAs) impurities in different
drug substances (N-nitroso-dimethylamine, N-nitroso-diethylamine, N-nitroso-ethyl-isopropyl
amine, N-nitroso-diisopropylamine, N-nitroso-dibutylamine, and N-nitroso-N-methyl-4-
aminobutyric acid). This method improves the sensitivity of the FDA method up to about 20
times, being also reduced the analysis time to the half. Its high sensitivity and easy sample
treatment should be of relevance in the field of pharmaceutical quality control for determining
NAs impurities associates to the synthesis processes. Moreover, for the detection of other possible
nitrosamines formed as by-products due to the modification of the manufacturing processes, a
screening method in full scan mode was also developed showing a deteccion limits similar to the
FDA.
Therefore, these two new methodologies provide a reliable method for routine control of
nitrosamine impurities in pharmaceutical industry.