title: Development of a method for the determination of cefovecin in plasma by HPLC.
abstract: A simple high-performance liquid chromatography method for the determination of cefovecin in small volume plasma has been developed. Following solid-phase extraction using Oasis HLB cartridges, samples were separated by reverse-phase high-performance liquid chromatography on an XBridge C8 (3.5 µm) 4.6 × 250 mm column and quantified using ultraviolet detection at 280 nm. The mobile phase was a mixture of 10 mm ammonium acetate (pH 3.5) and acetonitrile (89:11), with a flow rate of 0.85 mL/min. The standard curve ranged from 0.1 to 200 µg/mL. Intra- and Inter-assay variability for cefovecin was <10%, and the average recovery was >90%. The lower limit of quantitation was 0.1 µg/mL. This method was successfully applied to the analysis of cefovecin samples at our institution. This is also the first fully validated method with an internal standard that does not use mass spectrometry.
authors.affiliation: The University of Tennessee, College of Veterinary Medicine, Department of Biomedical and Diagnostic Sciences, 2407 River Drive, Knoxville, TN, 37996, USA.
authors.forename: Sherry
authors.initials: S
authors.lastname: Cox
authors.forename: Ali
authors.initials: A
authors.lastname: Martin
authors.forename: Mallery
authors.initials: M
authors.lastname: Hamill
authors.forename: Nealy
authors.initials: N
authors.lastname: Pistole
authors.forename: James
authors.initials: J
authors.lastname: Steeil
authors.forename: Juergen
authors.initials: J
authors.lastname: Schumacher
mesh_terms: D000818:Animals; D002511:Cephalosporins / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006737:Horseshoe Crabs; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012754:Sharks; D052616:Solid Phase Extraction; D013056:Spectrophotometry, Ultraviolet
chemical_list: D002511:Cephalosporins; C516253:cefovecin
keywords: UV detection; cefovecin; pharmacokinetics; solid-phase extraction