Related Experiment Video
Updated: Oct 5, 2025

A General Method for Detecting Nitrosamide Formation in the In Vitro Metabolism of Nitrosamines by Cytochrome P450s
Published on: September 25, 2017
Mass spectrometry based fragmentation patterns of nitrosamine compounds
Shardrack O Asare1, Jessica N Hoskins1, Richard A Blessing1
1Analytical Research and Development, AbbVie, 1 N. Waukegan Rd., North Chicago, IL, USA.
Rationale:
Nitrosamines are a class of mutagenic substances that can display high carcinogenic potential. New chemical entities may have the potential to form unique nitrosamines specific to the drug substance. It is therefore essential to understand the gas-phase fragmentation behavior of nitrosamine compounds to enable the development of analytical methods to characterize novel nitrosamine compounds.
Methods:
The gas-phase fragmentation behavior of eight model nitrosamine compounds representing the common substructures seen in many small molecule pharmaceutical compounds was studied with positive electrospray ionization tandem mass spectrometry (ESI-MS/MS). The fragmentation patterns of these compounds under various collision parameters available in commercially available mass spectrometers were studied.
Results:
Protonated nitrosamine compounds produced diagnostic fragment ions upon MS/MS. Three primary structure-dependent fragmentation pathways were observed. The first pathway involves the loss of 30 Da which corresponds to the loss of the NO radical from the protonated nitrosamine compound (Group 1). The second and third fragmentation pathways, which have not been reported for nitrosamine compounds, proceed via the loss of H2 O from the protonated nitrosamine compound (Group 2), and elimination and a loss of 46 Da (loss of NH2 NO) from the nitrosamine compound (Group 3).
Conclusions:
Results presented in this work provide an overview of the gas-phase fragmentation patterns of nitrosamine compounds and may be useful in identifying novel nitrosamine compounds in complex matrices.
Related Concept Videos
Mass Spectrometry: Amine Fragmentation
In amines, the number of nitrogen atoms affects the mass of the molecular ion, which is described by the nitrogen rule of mass spectrometry. This rule states that a compound containing...
Mass Spectrometry of Amines
Amines undergo fragmentation through α cleavage, producing nitrogen-containing cations—iminium ions—and alkyl radicals. Mass spectra of aromatic and cyclic aliphatic amines exhibit...
Mass Spectrometry: Carboxylic Acid, Ester, and Amide Fragmentation
For example,...
Mass Spectrometry: Molecular Fragmentation Overview
One type of fragmentation pattern is the cleavage of a single bond in the molecular ion. The cleavage leads to a radical and a cation. The cleavage can...
Mass Spectrometry: Aromatic Compound Fragmentation
Mass Spectrometry: Long-Chain Alkane Fragmentation

