Multiresidue Analytical Method for the Determination of Eight Penicillin Antibiotics in Muscle Tissue by Ion-Pair Reversed-Phase HPLC after Precolumn Derivatization

Abstract

Abstract A high-performance liquid chromatographic multiresidue method was developed for the determination of 8 penicillin compounds (benzylpenicillin, phenoxymethylpenicillin, ampicillin, amoxicillin, nafcillin, oxacillin, cloxacillin, and dicloxacillin) at trace levels in muscle tissue. This method involves extraction of the penicillins with phosphate buffer pH 9 followed by cleanup and concentration on a C18 solid¯phase extraction column and reaction with benzoic anhydride at 50°C for 5 min and with 1,2,4-triazole and mercury(II) chloride solution pH 9 at 65°C for 10 min. The derivatized compounds are eluted on a C18 column with a mobile phase containing acetonitrile and phosphate buffer (pH 6; 0.1 mol/L) loaded with sodium thiosulfate and ion-pairing tetrabutylammonium hydrogenosulphate. The method detection limit is approximately 3-11 μg/kg and the limit of determination was evaluated down to 25 μg/kg in line with the criteria of the EU decision No. 93/256/EEC.