Determination of cephalosporins in raw bovine milk by high-performance liquid chromatography

A high-performance liquid chromatographic (HPLC) method based on solid-phase extraction was developed for the determination of cefazolin, cefoperazone, cefquinome and ceftiofur in raw bovine milk. The milk fat was removed by centrifugation and the cephalosporins were extracted in acetonitrile. The extract was cleaned up by solid-phase extraction on an octadecyl sorbent. The compounds were separated by ion-paired gradient HPLC on a phenyl column with ultraviolet detection at 270 nm. The limits of detection estimated by a conservative model were 11 μg/kg for cefazolin and cefoperazone and 7 μg/kg for cequinome and ceftiofur. The mean recoveries were 86–88% for cefazolin, 91–93% for cefoperazone, 69–72% for cefquinome and 84–88% for ceftiofur in the concentration range 20–200 μg/kg.