Liquid chromatographic separation of fluoroquinolone antibacterials used as veterinary drugs

A liquid chromatographic method has been developed to separate a series of quinolones which are widely used as veterinary drugs: danofloxacin, difloxacin, enrofloxacin (and its metabolite ciprofloxacin), marbofloxacin, norfloxacin and sarafloxacin. Separation was performed by using an end-capped high-purity silica-based C8 column and a mobile phase consisting of acetonitrile-aqueous oxalic acid buffer solution. In order to establish the optimal conditions for an isocratic separation, a three-factor Doehlert experimental design was applied. Gradient elution was applied which reduced analysis time. Figures of merit of the method proposed, with fluorimetric detection, were evaluated.