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Multi-residual method detection and determination antibacterial substances in feed by LC-MS/MS technique

Wieloskładnikowa metoda wykrywania i oznaczania substancji przeciwbakteryjnych w paszach techniką LC-MS/MS

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Ewelina Patyra1, Carolina Nebot2, Rosa E. Gavilán2, Alberto Cepeda2, Krzysztof Kwiatek1

1Department of Hygiene of Animal Feedingstuffs, National Veterinary Research Institute, Pulawy, Poland

2Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Veterinary Medicine, University of Santiago de Compostela, 27002 Lugo, Spain

Key words: antibacterial substances, LC-MS/MS, feed, : Commission Decision 657/2002/EC

Słowa klucze: substancje przeciwbakteryjne, LC-MS/MS, pasza, Decyzja Komisji 657/2002/WE

The aim of this work was to develop a multi-component method allowing to analyze several antimicrobial substances from various chemical groups in non-target feeds in one LC-MS/MS analytical procedure.

The following compounds were selected for the study: ciprofloxacin, enrofloxacin, erythromycin, lincomycin, penicillin G, tiamulin, trimethoprim, tylosin, sulfadiazine, sulfamethazine and valnemulin. From two grams feed sample extraction of these compounds was made using a mixture of 0.1% formic acid in acetonitrile. The samples were shaken and then centrifuged, 4 ml of the obtained extract was concentrated under a stream of nitrogen. Residues was dissolved in Milli-Q water and filtered through a syringe filter and then analyzed with the use LC-ESI-MS/MS. The analyzes were carried out on a biphenyl column using a mobile phase consisting of 0.1% formic acid in acetonitrile and 0.1% formic acid in water Mili-Q. The analysis time was 24 minutes.

The presented method was validated in accordance with the guidelines of the European Commission Decision 657/2002/EC. Good validation parameters were obtained with high recoveries ranging from 76.04 to 117.39% and coefficients of variation ranging from 2.41 to 19.27%. The developed method allows for the determination of eleven antimicrobial compounds ranging from 250 to 2500 μg/kg of feed and is useful for routine feed analysis of various origins.


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