Marbofloxacin is a veterinary fluoroquinolone drug used for the treatment of bacterial infections in animals. In this study, a stability-indicating UV-visible spectrophotometric method was developed and validated for the estimation of marbofloxacin in bulk and their dosage form. The method involved the use of a UV-visible spectrophotometer to measure the absorbance of marbofloxacin at its maximum wavelength of 301 nm. The method was validated for linearity, accuracy, specificity, precision, and robustness as per ICH guidelines. The linearity range was found to be 5-50 μg/mL, with a correlation coefficient of 0.9996. The percentage recovery was found to be within the limit of 98-102%, indicating the accuracy of the method. The method was found to be specific, as there was no interference from the excipients or degradation products of marbofloxacin. The precision of the method was evaluated by repeatability and intermediate precision, and the results showed RSD less than 2%. The method was found to be robust, and small deliberate changes were made to the analytical variables, which did not significantly affect the method's performance. The validated method was successfully applied for the estimation of marbofloxacin in bulk and pharmaceutical dosage form. The proposed method is a simple, accurate, precise, and stability-indicating UV-visible spectrophotometric method for the determination of marbofloxacin in bulk and pharmaceutical formulations.