Isocratic High Performance Liquid Chromatography Assay for Quantification of Ceftiofur Hydrochloride in Bubaline...

We optimized and validated an isocratic high-performance liquid chromatography (HPLC) assay for quantification of ceftiofur hydrochloride in bubaline plasma. Ceftiofur, its metabolic products and protein-bound residues were cleaved, derivatized into desfuroylceftiofur acetamide and injected into HPLC system. The mobile phase comprising of sodium dihydrogen phosphate (0.025 M, pH 7) and acetonitrile (34:66, v/v), was driven at a flow rate of 1 mL/min, and separation was achieved using C18 column. Isocratic elution was performed with an injection volume of 45 µL and analyte was scanned at 310 nm. The linearity range, limit of detection and limit of quantification were 0.1-10 µg/mL, 0.03 µg/mL and 0.11 µg/mL respectively. Moreover, the accuracy, precision and recovery remained within the acceptable limits. The assay was effectively applied for determining the concentration of ceftiofur in plasma samples collected from ceftiofur-treated buffalo calves.