The prevalent use of antibiotics, e. g. sulphonamides, tetracyclines, beta-lactams, macrolides, aminoglycosides, etc., in dairy farming to prevent and treat microbial infections e. g. mastitis, can be a potential source of veterinary drug residues in milk. Antibiotic residues constitute a risk to human health, since they can cause allergic reactions in hypersensitive individuals or they may lead to the appearance of drug-resistant bacteria. Analysis of these residues plays a key role in ensuring food safety. Regulatory agencies in the European Union and in other countries have established maximum residue limits and requirements concerning the performance of analytical methods and the interpretation of the results. This review describes the state of the art in the analytical strategies concerning the multi-class as well as the multi-residue analysis of antibiotics in milk. Since milk is a complex matrix due to its high protein and fat content, which often interfere in analytical procedures, special focus has been placed on sample preparation: extraction and clean-up. Confirmation of antibiotic residues according to European Decision 657/2002/EC has been also discussed.
A high-performance liquid chromatographic method was developed for the determination of five penicillins: penicillin G (PENG), penicillin V (PENV), oxacillin (OX), cloxacillin (CLO), and dicloxacillin (DICLO), in bovine muscle. Samples were macerated with a mixture of H(2)O/CH(3)CN (1:1) and purified using RP-8 Adsorbex SPE cartridges after centrifugation, with mean recovery from spiked samples higher than 89%. The separation of the examined penicillins was achieved on an analytical column, an Inertsil C8 5 microm, 250x4 mm(2), at ambient temperature. The mobile phase consisted of 0.1% TFA/ACN 50:50 v/v delivered isocratically at a flow rate of 1.1 mL/min. Analytes were monitored at 240 nm. The procedure was validated according to the European Union Decision 2002/657/EC by means of selectivity, stability, decision limit, detection capability, accuracy, and precision. Method's LOQ values achieved were 54 microg/kg for PENG and DICLO, 46 microg/kg for PENV, 16 microg/kg for OX, and 43 microg/kg for DICLO. The detection capabilities (CC(beta)) were 73.6 microg/kg for PENG, 29.1 microg/kg for PENV, 350.6 microg/kg for OX, 379.9 microg/kg for CLO, and 355.8 microg/kg for DICLO. The method was applied to various samples from the local market. Two penicillins were identified by photodiode array (PDA) detection and quantified.
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