Simultaneous liquid chromatographic determination of some tetracyclines in serum

A sensitive liquid chromatographic method has been developed for the simultaneous determination of oxytetracycline, minocycline, tetracycline, and doxycycline in serum. A serum sample is vortex-mixed with a solution of mobile phase for tetracyclines and 2% (v/v) phosphoric acid. The mixture is filtered using a 30,000 molecular weight cutoff microseparation tube which separates high-molecular-weight solutes following low-speed centrifugation. Tetracyclines are separated from other serum components by reverse phase liquid chromatography (LC) with buffered methanol mobile phase. Ultraviolet absorbance of the column effluent is monitored at 267 nm. Concentrations as low as 0.2 micrograms/mL of tetracyclines in serum are quantitatable, with recoveries from 76.2 to 102.6% and coefficients of variation from 2.69 to 5.36%. The method has been tested in bovine, porcine, equine, caprine, ovine, canine, feline, and avian (turkey) serum.