A method for determination of oxytetracycline, tetracycline and chlortetracycline residues in milk by high performance liquid chromatography was optimized and validated. Linearity, selectivity, trueness, precision, limits of detection and quantitation were evaluated by in-house validation procedures employing reference materials, sample blanks and spiked samples. Milk samples were homogenized, treated with sodium succinate pH 4.0 buffer and centrifuged, discarding the precipitate. Tetracyclines were removed from the supernatant by chelating with metal ions bound to a chelating resin and eluted with McIlvaine-EDTA pH 4.0 buffer. Mean recoveries for tetracyclines ranged from 76.2 to 104.9% with coefficients of variation from 1.4 to 18.9%. Detection limits of 10, 15 and 40ng/mL and quantitation limits of 10, 20 and 50ng/mL were determined for oxytetracycline, tetracycline and chlortetracycline, respectively.
milk; residues; tetracyclines; high performance liquid chromatography
