This study aimed to develop a simple stability-indicating HPLC-DAD method for the assay of Tildipirosin (TPS), which is a 16-member ring semi-synthetic macrolide, in an injectable solution. TPS is commonly used in the USA for the treatment of animal's respiratory infection. The isocratic elution was set up using the Inertsil-ODS column with the flow rate set at 0.9 mL min-1 and the detection wavelength of 285 nm. The column temperature was ambient and the overall runtime was 5 minutes. Additionally, the mobile phase was water: acetonitrile: trifluoroacetic acid (53.45: 46.45: 0.10, by volume). The HPLC-forced degradation stability method for TPS in Egydipiro Injectable Solution was performed in different stress conditions, i.e. light/UV, heat, acid, base, and oxidation. The results showed the linearity range was 36.00 to 108.00 µg mL-1 for TPS. The correlation coefficient was higher than 0.999 and the accuracy was 98.78%. The degradation percentages were presented ranging from 31.54% in the photodegradation condition to 50.10% in the base degradation. The Eco-scale score was about 82 points indicating an excellent green chromatographic technique. Therefore, the novel stability method was unique in terms of simplicity and environment-friendly aspects. The new HPLC–DAD method was the first, green, stability-indicating chromatographic method for a TPS analysis with its degradation products. However, the major drawbacks of the new chromatographic method are the incomplete identification of degradation products and the inability to propose an appropriate mechanism for drug degradation. Overall, the method can be employed to evaluate TPS stability under various storage conditions.
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