Determination of cloxacillin in milk and blood of dairy cows by high-performance liquid chromatography.

A rapid and sensitive high-performance liquid chromatographic method is described for the determination of cloxacillin residues in milk and serum. Only a clean-up step after deproteinization is necessary before the analysis. The chromatographic system involves the use of a C18 column and ultraviolet absorbance detection at 225 nm. The mobile phase was acetonitrile-0.02 M KH2PO4 (21:79) at pH 5. Recoveries for cloxacillin were 83.5 and 75.7% in serum and milk, respectively. Detection limits (10 ng/ml in milk and 50 ng/ml in serum) were below the stipulated European Union maximum residue limits for cloxacillin. Thus, the described method showed the same accuracy, precision and sensitivity as the microbiological assays but without interferences caused by other drugs commonly used in therapy. Analysis of different blood and milk samples obtained at different times from dairy cows treated with an intramammary dose of cloxacillin benzatine showed undetectable cloxacillin levels both in milk and blood samples.