Abstract
In this study, an improved liquid chromatographic (LC) method for the analysis of tylosin and its impurities has been developed. A Kinetex EVO C18 column (150 × 4.6 mm, 2.6 μm) packed with superficially porous particles was used as stationary phase. Gradient elution was applied with two mobile phases (A and B) containing acetonitrile, water and 0.2 M ammonium acetate at different ratios (20:10:70 (v/v/v) for A and 60:10:30 (v/v/v) for B). This volatile mobile phase enables the method to be coupled with mass spectrometry (MS) for additional detection and characterization of tylosin impurities. Selectivity, sensitivity, linear calibration, accuracy, precision and robustness of this analytical method were assessed through method validation. In addition, impurities above 0.05% were characterized via LC–MS/MS. It is the first time that a MS compatible method for analysis of tylosin and its impurities is presented. Moreover, it shows a considerably shorter analysis time than previously published methods.
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