Study of the impurity profile and polymerized impurity in mezlocillin sodium by the multiple heart‐cutting two‐dimensional liquid chromatography coupled with ion trap/time‐of‐flight mass spectrometry

RATIONALE: Eleven impurities and one polymerized impurity in mezlocillin were identified and their formation mechanisms were investigated in this study. The sources and reasons for the formation of impurities were revealed, which may guide industry to improve the manufacturing process and storage conditions and reduce the content of impurities in products. The results from this study also provided a scientific basis for the improvement of official monographs in pharmacopoeias. METHODS: The impurity profiles and polymerized impurity in mezlocillin were studied by multiple heart-cutting two-dimensional liquid chromatography coupled with ion trap time-of-flight mass spectrometry (2D-LC/IT-TOF MS) in both positive and negative modes of electrospray ionization. Target eluents from the first dimensional chromatography with a non-volatile mobile phase were trapped and sent to the second dimensional chromatography with a volatile mobile phase by a switching valve. The structures of the impurities in the mezlocillin drug substance were deduced based on the high-resolution MSn data. RESULTS: In the environment of water, oxygen, high temperature, acid and base, a series of degradation products could be easily produced from mezlocillin. Mezlocillin was hydrolyzed into impurities I, IV, V and X, and was degraded into impurity III by methanolysis. Mezlocillin was oxidized into sulfoxide by producing impurity XI. Furthermore, impurities VI, VII, VII and IX were all isomers of mezlocillin. The proposed formation pathways of these products were demonstrated in this study. CONCLUSIONS: Eleven degradation impurities and one polymerized impurity in mezlocillin were separated and characterized. Based on characterization of impurities, this study discovered the mechanism of impurity production and provided guidance for manufacturers to improve the process and storage conditions and reduce levels of impurities.