A Simple HPLC Method for Detection of Fluoroquinolones in Serum of Avian Species

A high-performance liquid chromatography (HPLC) method with fluorescent detection was validated to analyze fluoroquinolones in small volumes of serum samples from avian species. A reversed-phase column was used. Excitation and emission wavelengths for ciprofloxacin and enrofloxacin were set at 277 nm and 418 nm, respectively. These parameters for danofloxacin were 295 nm and 500 nm and for marbofloxacin were 338 nm and 425 nm, respectively. The mobile phase consisted of acetonitrile and phosphate buffer at a flow rate of 1 mL/min. The standard curves for the tested fluoroquinolones were linear over a range 10–1000 ng/mL. Enrofloxacin did not interfere with ciprofloxacin, danofloxacin, and marbofloxacin. It was used as an internal standard for danofloxacin and marbofloxacin detection. The inter-assay precision varied between 1.73 and 13.21%. Values for intra-assay precision were between 1.11 and 7.18%. Accuracy was higher than 90% at all of the tested concentrations and compounds. Mean recovery was higher than 85% for ciprofloxacin and enrofloxacin and >95% for danofloxacin and marbofloxacin.