Cracking the EI Code: Fragmentation Patterns for Structural Elucidation of Neonicotinoid Insecticides by GC-MS

Although neonicotinoids (NEOs) insecticides have been extensively detected in environmental matrices, their electron ionization (EI) fragmentation patterns have never been systematically elucidated. Because of the poor thermal stability of NEOs, gas chromatography-mass spectrometry (GC-MS) is rarely used in analysis of NEOs. This work investigated eight representative NEOs by GC-MS using multiple isotopically labeled standards, including ²H and ¹³C labeled standards to clarify fragmentation routes and establish robust structural assignment criteria. We found two main factors affecting EI fragmentation patterns. (1) The pharmacophore controlled the backbone cleavage pathway. Specifically, nitroguanidines underwent transketolation with loss of N₂O, while cyanoamidines fragment via α-cleavage. (2) The heterocycle moiety determined the diagnostic fragment ion. A heterocyclic group with chloropyridine gave the diagnostic fragment ion of m/z 126, and a chlorothiazole moiety gave the diagnostic fragment ion of m/z 132. The lack of m/z 126 and m/z 132 ions in the case of dinotefuran (DNT), which contained neither of these two moieties, indirectly suggested the rules. This set of rules effectively bridged the gap in EI mass spectral interpretation, providing a basis for ion transition selection and fragment assignment during GC-MS method development. Moreover, it could provide supportive mass-spectrometric clues for potential application in the structural elucidation of related compounds.