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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

s. 38 - 39

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.
 

References

  1. Anon: Commission Decision 2002/657/EC. 2002. of 12 August implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. OJ. L 221:8–36.
  2. Borras S, Companyo R, Granados M, Guiteras J, Perez-Vendrell AM, Brufau J, Medina M, Bosch J. 2011. Analysis of antimicrobial agents in animal feed. Trends Anal Chem. 30:1042–1064.
  3. Gavilán R.E., Nebot C., Patyra E., Miranda J.M., Franco C.M., Cepeda A. 2018.  Simultaneous analysis of coccidiostats and sulfonamides in non-target feed by HPLC-MS/MS and validation following the Commission Decision 2002/657/EC. Food Additiv. Contam. Part A.. doi: 10.1080/19440049.2018.1466061.
  4. Gavilán RE, Nebot C, Veiga-Gomez M, Roca-Saavedra P, Vazquez Belda B, Franco CM, Cepeda A. 2016. A confirmatory methods based on HPLC-MS/MS for the detection and quantification of residue of tetracyclines in nonmedicated feed. J Anal Methods Chem. doi:10.1155/2016/1202954.
  5. Kaklamanos G, Vincent U, von Holst C. 2013. Analysis of antimicrobial agents in pig feed by liquid chromatography coupled to orbitrap mass spectrometry. J Chromatogr A. 1293:60–74.
  6. Patyra E, Kwiatek K. 2017. Development and validation of multi-residue analysis for teteracycline antibiotics in feed by high performance liquid chromatography coupled to mass spectrometry. Food Additiv Contam Part A. 34 (9):1553–1561.
  7.  Patyra E, Kwiatek K. 2017. Determination of fluoroquinolones in animal feed by ion pair high-performance liquid chromatography with fluorescence detection. Anal Letters. doi:10.1080/00032719.2016.1249876.