The aim of this study was to develop and validate a simple high performance liquid chromatography (HPLC) method for the determination of cefovecin sodium in small volume of lion’s plasma. Analytical separation was obtained in less than 5 minutes (4.29-4.42 min) using a C-18 column with UV detection at λmax 254nm. The extraction of cefovecin sodium was carried out in methanol and acetonitrile. The mobile phase was composed of water: acetonitrile: methanol in a ratio of 60:20:20 at pH 3.1. The method was validated according to FDA Bioanalytical assay guidance. The limit of detection of cefovecin sodium was 0.05 ug/ml, and the limit of quantification was 0.1ug/ml. The range of linearity consisted of 5 points with 3 runs for each point that result in excellent linearity with R2 = 0.9998. The recoveries were 98-99.25 ± %RSD (NMT 2%). The intra-day and inter-day precision (%CV) of this method was 2.3-10.26% ± SD and 2.44-7.22% ± SD respectively. A straightforward, easy and successful HPLC method was developed that determined and quantified cefovecin sodium concentrations in African Lion's plasma that can be used in the future work in a pharmacokinetic study of cefovecin sodium.

Sultan M. Alshahrani and John Mark Christensen