Structural analysis of photo-degradation in thiazole-containing compounds by LC–MS/MS and NMR

The photo-degradation behavior of a pharmaceutical compound previously under development for treatment of overactive bladder was studied. Samples of {4-(4-chloro-3-fluorophenyl)-2-[4-(methyloxy)phenyl]-1,3-thiazol-5-yl} acetic acid were stressed with visible light and were observed to degrade into a single primary photo-degradation product. This unknown product was analyzed by liquid chromatography tandem mass spectrometry (LC–MS/MS) with accurate mass measurement and hydrogen/deuterium exchange to determine its molecular weight and formula, isotope distribution patterns and exchangeable protons, and product ion structures. By comparison of the fragmentation pathways of the protonated and sodiated species, the charge was found to locate in the electron-rich part of the molecule after fragmentation. MS-derived structural information combined with stopped-flow 1H LC-nuclear magnetic resonance (NMR) analysis suggested that the degradation product was 4-chloro-N-(4-methoxybenzoyl)-3-fluorobenzamide. This unique photo-degradation product was subsequently isolated using preparative-scale chromatography, and its structure was confirmed using 1D and 2D NMR techniques involving the 1H, 13C, 15N and 19F nuclei. The structure of this product suggests that {4-(4-chloro-3-fluorophenyl)-2-[4-(methyloxy)phenyl]-1,3-thiazol-5-yl} acetic acid has reacted with singlet oxygen (1Δg) via a [4 + 2] Diels-Alder cycloaddition upon photo-irradiation to cause photo-oxygenation in the solid-state (as is common in solution phase), resulting in an unstable endoperoxide that rearranges to the final degradation product structure. Photo-degradation of a structurally related thiazole, 4-(4-Chlorophenyl)thiazol-2-amine, proceeded via a similar process but in a less reactive manner. However, when exposed to the same conditions, sulfathiazole did not degrade, indicating that this photo-degradation process may only occur for thiazole-containing compounds with specific substituents, such as aryl rings.