Sensitive Quantification of N-Nitroso Dibutylamine Impurities in Tramadol Tablets Using Optimized LC-MS/MS Method

Background: NDBA, a recognized genotoxic and carcinogenic impurity in pharmaceutical products, is employed as a model compound. As a result, regulatory agencies worldwide have imposed stringent testing requirements for Nitrosamine impurities, and there is a demand for sensitive and selective analytical methods for their detection. Objective: The purpose of this study was to develop and validate a Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) method for the accurate determination of NDBA impurities in Tramadol tablets, thereby overcoming the limitations of High Performance Liquid Chromatography (HPLC) in identifying low-level impurities. Methods: Selectivity and sensitivity of the LC-MS/MS method were optimized, and the Limit of Quantitation (LOQ) was set to 5% and the Limit of Detection (LOD) to 3% of the specification limit. The linearity was characterised over a range of 5% to 150% relative to the specification limit. At several concentration levels within the analytical range, the precision and accuracy were determined. discussion: The blank chromatogram provides a baseline to look for interfering peaks at the N-nitroso dibutylamine (NDBA) impurity retention time. In this study, the blank chromatogram contained no signals at the NDBA retention time (3.4 min) demonstrating that the method is very specific. This is important as this demonstrates the method’s ability to distinguish the NDBA impurity from the other matrix or mobile phase components. Specificity is one of the cornerstone components for an analytical method for genotoxic impurities quantification with no interference Results: In the specified range, the developed method was found to be linear (R² > 0.999). Low relative standard deviation precision studies yielded results consistent with those obtained with this analytical method, and accuracy assessments demonstrated that the method could quantitatively measure NDBA over a range of concentrations. The method was suitable for routine quality control and met and exceeded regulatory performance requirements (LOQ and LOD). Conclusion: It was established that a validated LC-MS/MS method provides a reliable, sensitive, and selective approach for determining NDBA impurities in Tramadol tablets. The use of this method enables compliance with the most stringent regulatory standards for pharmaceutical product safety and efficacy.