Orbifloxacin is a fluoroquinolone antibiotic which inhibits bacterial growth by inhibiting DNA gyrase and topoisomerase IV.

Orbifloxacin has very limited non-medical applications, as it is primarily known and used as a veterinary antibiotic for the treatment of bacterial infections in animals. However, it can be used as a reference standard in analytical chemistry for the development and validation of methods for the detection and quantification of fluoroquinolone antibiotics, such as Orbifloxacin, in various samples, such as food, environmental, and biological samples. Orbifloxacin has been used as a standard for the analysis of fluoroquinolone antibiotics in food products, such as meat, milk, and eggs, as well as in environmental samples, such as water and soil. Moreover, its potential use in animals, particularly livestock and poultry, highlights the importance of prudent use and management of the antibiotic to prevent the development and spread of antibiotic-resistant bacteria.

Orbifloxacin inhibits bacterial DNA synthesis through interference with the activity of DNA gyrase and topoisomerase IV enzymes, which are necessary for the replication, transcription, and repair of bacterial DNA. Orbifloxacin is active against Gram-negative and Gram-positive bacteria, including many strains of Escherichia coli, Pasteurella, Streptococcus, and Staphylococcus species.