Development and validation of a HPLC method for the determination of oxytetracycline and enrofloxacin in veterinary formulation

摘要

An isocratic reversed phase high performance liquid chromatographic method with DAD detection was developed for the analysis of oxytetracycline and enrofloxacin. The mobile phase consisted in acetonitrile: methanol: 0.4% orthophosphoric acid adjust to pH 2.7 with triethanolamine (20:10:70). The UV detection was carried out at 254 nm, and the flow rate was 1.0 mL/min. The separation was carried out on a Nucleosil C18 column, 5 μm 250 mm x 4.6 mm, which was maintained at 20 0C. This method was validated by specificity parameters, linearity, limits of detection and quantification, precision and accuracy.