Assay of afoxolaner and determination of its related substances in commercial bulk batches of afoxolaner by reversed-phase HPLC method based on a short octadecyl column

•First reported RP-HPLC method to analyze afoxolaner and its related substances.•First reported stability-indicating method to analyze afoxolaner and its related substances.•Application of rapid and automated chromatographic method development tools.•The method was successfully validated according to the ICH guidelines.•This proposed method is simple, efficient, sensitive, and QC-friendly for routine analysis of afoxolaner.