Background: In this work, we developed and validated a simple, rapid and accurate reverse-
HPLC for the multi-determination of enrofloxacin (EFX), ciprofloxacin (CFX), and oxytetracycline
(OXY) in raw materials and veterinary pharmaceutical formulations.
Method: Optimum chromatographic separations among these three analytes have been achieved within
3 min employing an Agilent Poroshell 120 EC-C18 column (100 mm × 3.0 mm, 2.7 µm) as the stationary
phase using a mobile phase composted of water:acetonitrile (87: 13, v/v) plus 0.1% trifluoroacetic
acid, flow rate at 3 mL min-1 and detection at 280 nm.
Result: The method resulted in excellent separation (Rs > 3) and good efficiency (theoretical plates >
5000) for all of the analytes. The method showed acceptable results in the procedure of validation according
with ICH and USP guidelines with respect to selectivity, linearity, limits of detection and quantitation,
accuracy, precision and robustness. Finally, the developed method can be applied in the determination
of EFX, CFX and OXY in routine quality control of raw materials and veterinary pharmaceutical