DEVELOPMENT OF A HPLC METHOD FOR THE DETERMINATION OF ENROFLOXACIN AND CIPROFLOXACIN IN REPTILE PLASMA AFTER TRANSDERMAL DELIVERY

Iga Czyz, Jean-Paul Salvi, Alban Ducrotte, Plamen Kirilov, Sébastien Perrot, Charly Pignon, Fabrice Pirot, Françoise Falson, Roselyne Boulieu*

A rapid and simple high-performance liquid chromatography method using diode array detection for determination of enrofloxacin and ciprofloxacin in snake’s plasma was developed. Several snakes were treated transdermally with a single dose of enrofloxacin emulsions corresponding to an antibiotic dose of 5 mg/kg. Enrofloxacin and ciprofloxacin were analyzed using a core-shell silica particle stationary phase (Kinetex® RP–C18, 150 × 4.6 mm, particle size 5 μm) and 0.002 M phosphoric acid/acetonitrile (83:17, v/v) as mobile phase. Calibration curves were linear over the concentration range of 2–100 μM and 3–100 μM. The intra-day and inter-day coefficient of variations were below or equal to 10% for both compounds. Limits of quantification for enrofloxacin and ciprofloxacin were 2 μM and 3 μM respectively. Sample treatment procedure consisted of deproteinization with perchloric acid. The described HPLC method using core-shell silica particles results in better resolution, higher sensitivity and low back pressure.