ORIGINAL RESEARCH article
Front. Anal. Sci.
Sec. Pharmaceutical analysis
A case study on N-nitrosamine investigation in a marketed drug: From analytical procedure validation to risk mitigation decision
Provisionally accepted
- Universite de Liege, Liège, Belgium
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N-nitrosamines, a class of organic compounds recognized for their mutagenic and carcinogenic properties, have been raising major public health concerns following their detection in pharmaceuticals since 2018. These impurities can form during manufacturing processes, particularly through the interaction of amines with nitrosating agents. Regulatory bodies have launched procedures mandating a systematic investigation of medicinal products for N-nitrosamine contamination. This process involves risk evaluation, confirmatory testing, and the implementation of risk mitigation measures. Hyphenated analytical techniques, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS), have become the standard for N-nitrosamine detection thanks to their high sensitivity and specificity. In this work, based on a previously developed Quantitative Structure Retention Relationship approach for the determination of N-nitrosamines, sensitive LC-MS/MS methods were proposed and validated in accordance with the ICH Q2(R2) guidelines as limit tests for trace-level detection of N-nitrosamines. These methods were applied in confirmatory testing, which demonstrated the absence of most N-nitrosamines throughout the product's shelf life, except for one potentially problematic impurity that nevertheless remained below the established acceptable limit, thus requiring routine quantitative analysis. This finding highlights the need for continued monitoring through routine analysis to ensure the safety and quality of pharmaceutical products.
Keywords: Analytical procedure validation, drug safety, impurity, LC-MS/MS, N-nitrosamine, pharmaceuticals, Quality control
Received: 21 Dec 2025; Accepted: 13 Feb 2026.
Copyright: © 2026 Zhang, Houari, Van Laethem, Kumari, ZIEMONS, Hubert and Hubert. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Yue Zhang
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