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A liquid chromatographic method with UV detection at 325 nm was developed for simultaneous determination of amoxicillin, ampicillin, penicillin G, and cloxacillin residues in bovine muscle tissue as their mercaptide derivatives. The penicillins are extracted from bovine tissues with 0.1 M phosphate buffer (pH 8.5), cleaned up on a t-C18 Sep-Pak cartridge, and eluted with 2 ml acetonitrile. After the acetonitrile in the eluate is evaporated to dryness, the residue is dissolved in 200 |µL (40 + 60, v/v) aceton itrile-phosphate buffer (pH 6.5) and derivatized with acetic anhydride and mercuric chloride in the presence of 1,2,4-triazole at 65°C for 30 min. Gradient analysis on a Spherisorb 5 µm ODS(2) (octadecyl silane) analytical column using a binary mobile phase consisting of acetonitrile and 0.10M phosphate buffer (pH 6.5) in the presence of 0.0157M sodium thiosulfate at 1 mL/min permits determination of each intact penicillin in bovine muscle tissue at ≥10 ppb with recoveries ≥72%. This laboratory method provides detection sensitivities equivalent to those of rapid tests used for screening β-lactam drug residues in bovine tissue samples for regulatory enforcement.

Multiresidue Liquid Chromatographic Method for Determining Residues of Mono- and Dibasic Penicillins in Bovine Muscle Tissues