Quantitative analysis of residual tetracyclines in buffalo meat using RP-HPLC

Abstract

A liquid chromatographic method is described for the simultaneous quantitative measurement of oxytetracycline and chlortetracycline in buffalo meat. After extraction of the meat samples using Mcllvaine buffer – EDTA solution and SPE clean up, the analysis of the antimicrobials was carried out using a reverse phase HPLC using UV detector. The chromatographic separation was accomplished with mobile phase consisting of 0.1M oxalic acid, methanol and acetonitrile (70:15:15, v/v) in an isocratic elution mode. The volume of injection and flow rate of the mobile were 1.5 ml/min and 50 μl respectively.UV detector was operated at a wavelength 360nm. Linearity, recovery, selectivity, limit of detection, limit of quantification was evaluated in buffalo meat matrix at drug concentrations ranging from 25–1000 ng/g. Mean extraction recoveries of oxytetracycline and chlortetracycline were in the range of 90–97%. The limits of quantification for oxytetracycline and chlortetracycline were 38.30 and 36.63 μg/kg, respectively. The proposed RP-HPLC method is quite adequate for routine analysis of residual tetracyclines in buffalo meat below Codex MRL of 200 μg/kg.