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Determination of Coccidiostats in Milk Products by LC–MS and Its Application to a Fermentation Experiment

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Abstract

A liquid chromatographic tandem mass spectrometric (LC–MS–MS) method for the determination of five chemical coccidiostats (decoquinate, diclazuril, halofuginone, nicarbazin, and robenidine) and five ionophore coccidiostats (maduramicin, monensin, narasin, salinomycin, and semduramicin) in yoghurt, kefir, and sour cream is presented. Lasalocid, the sixth ionophore listed in 124/2009/EC was not included because of its extremely dissimilar behavior during sample preparation. Main steps of the method include extraction with acetonitrile, centrifugation, clean-up on Oasis HLB solid phase extraction cartridge, evaporation under nitrogen stream, and LC–MS–MS determination. Selectivity, linearity, sensitivity, accuracy, repeatability, within-laboratory reproducibility, limit of determination, and limit of quantitation were determined during the validation procedure. The method proved to be applicable for both qualitative and quantitative determination of the ten above-mentioned target compounds. In our in-house fermentation experiments, milk fortified with coccidiostats was fermented to get yoghurt, kefir, and sour cream. Our results show that the coccidiostat content did not change significantly during fermentation for any of the target compounds.

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References

  1. Commission Regulation 1831/2003/EC (2003) Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition. Off J Eur Union L268:29–43

    Google Scholar 

  2. (2003) Community Register of Feed Additives to Regulation (EC) No. 1831/2003. List of the authorized additives in feedingstuffs (1) published in application of Article 9t (b) of Council Directive 70/524/EEC concerning additives in feedingstuffs. Off J Eur Union C50:1–144

  3. Commission Regulation 124/2009/EC (2009) COMMISSION REGULATION (EC) No 124/2009 of 10 February 2009 setting maximum levels for the presence of coccidiostats or histomonostats in food resulting from the unavoidable carry-over of these substances in non-target feed. Off J Eur Union L40:7–11

    Google Scholar 

  4. Huebra MJG, Vincent U, von Holst C (2010) Determination of semduramicin in poultry feed at authorized level by liquid chromatography single quadrupole mass spectrometry. J Pharm Biomed Anal 53:860–868

    Article  Google Scholar 

  5. Feás X, Vázquez BI, Iglesias A, Cantalapiedra J, Fente CA, Cepeda A (2010) Sensitive determination of decoquinate in milk by high-performance liquid chromatographic coupled to laser-induced fluorescence detection. Food Anal Methods 3:138–143

    Article  Google Scholar 

  6. Matabudul DK, Crosby NT, Lumley I, Sumar S (2001) The optimisation of a rapid method for the determination of lasalocid in poultry feed using supercritical fluid extraction and high performance liquid chromatography. Food Chem Anal Nutr Clin Methods Sect 75:465–471

    CAS  Google Scholar 

  7. Matabudul DK, Conway B, Lumley ID (2000) A rapid method for the determination of lasalocid in animal tissues and eggs by high performance liquid chromatography with fluorescence detection and confirmation by LC–MS–MS. Analyst 125:2196–2200

    Article  CAS  Google Scholar 

  8. Dusi G, Faggionato L, Gamba V, Baiguera A (2000) Determination of nicarbazin and clopidol in poultry feeds by liquid chromatography. J Chromatogr A 882:79–84

    Article  CAS  Google Scholar 

  9. Chéneau E, Henri J, Pirotais Y, Abjean J-P, Roudaut B, Sanders P, Laurentie M (2007) Liquid chromatography–electrospray tandem mass spectrometric method for quantification of monensin in plasma and edible tissues of chicken used in pharmacokinetic studies: applying a total error approach. J Chromatogr B 850:15–23

    Article  Google Scholar 

  10. Mortier L, Daeseleire E, Peteghem CV (2005) Liquid chromatographic tandem mass spectrometric determination of five coccidiostats in poultry eggs and feed. J Chromatogr B 820:261–270

    Article  CAS  Google Scholar 

  11. Mortier L, Daeseleire L, Peteghem CV (2005) Determination of the coccidiostat diclazuril in poultry feed and meat by liquid chromatography–tandem mass spectrometry. Anal Chim Acta 529:229–234

    Article  CAS  Google Scholar 

  12. Yakkundi S, Cannavan A, Elliott CT, Lövgren T, Kennedy DG (2003) Development and validation of a method for the confirmation of halofuginone in chicken liver and eggs using electrospray tandem mass spectrometry. J Chromatogr B 788:29–36

    Article  CAS  Google Scholar 

  13. Cannavan A, Ball G, Kennedy G (1999) Determination of nicarbazin in feeds using liquid chromatography–electrospray mass spectrometry. Analyst 124:1431–1434

    Article  CAS  Google Scholar 

  14. Mortier L, Daeseleire E, Delahaut P (2003) Simultaneous detection of five coccidiostats in eggs by liquid chromatography–tandem mass spectrometry. Anal Chim Acta 483:27–37

    Article  CAS  Google Scholar 

  15. Vincent U, Ezerskis Z, Chedin M, von Holst C (2011) Determination of ionophore coccidiostats in feeding stuffs by liquid chromatography-tandem mass spectrometry. Part II. Application to cross-contamination levels and non-targeted feed. J Pharm Biomed Anal 54:526–534

    Article  CAS  Google Scholar 

  16. Thompson TS, Noot DK, Kendall JD (2011) Determination of ionophores in raw bovine milk using LC–MS/MS: application to residue surveillance. Food Chem 127:321–326

    Article  CAS  Google Scholar 

  17. Vincent U, Chedin M, Yasar S, von Holst C (2008) Determination of ionophore coccidiostats in feedingstuffs by liquid chromatography–tandem mass spectrometry. Part I. Application to targeted feed. J Pharm Biomed Anal 47:750–757

    Article  CAS  Google Scholar 

  18. Matabudul DK, Conway B, Lumley I, Sumar S (2001) The simultaneous determination of the ionophore antibiotics in animal tissues and eggs by tandem electrospray LC–MS–MS. Food Chem 75:345–354

    Article  CAS  Google Scholar 

  19. Dusi G, Gamba V (1999) Liquid chromatography with ultraviolet detection of lasalocid, monensin, salinomycin and narasin in poultry feeds using pre-column derivatization. J Chromatogr A 835:243–246

    Article  CAS  Google Scholar 

  20. Shao B, Wu X, Zhang J, Duan H, Chu X, Wu Y (2009) Development of a rapid LC–MS–MS method for multi-class determination of 14 coccidiostat residues in eggs and chicken. Chromatographia 69:1083–1088

    Article  CAS  Google Scholar 

  21. Dubreil-Chéneau E, Bessiral M, Roudaut B, Verdon E, Sanders P (2009) Validation of a multi-residue liquid chromatography–tandem mass spectrometry confirmatory method for 10 anticoccidials in eggs according to Commission Decision 2002/657/EC. J Chromatogr A 1216:8149–8157

    Article  Google Scholar 

  22. Galarini R, Fioroni L, Moretti S, Pettinacci L, Dusi G (2011) Development and validation of a multi-residue liquid chromatography-tandem mass spectrometry confirmatory method for eleven coccidiostats in eggs. Anal Chim Acta 700:167–176

    Article  CAS  Google Scholar 

  23. Olejnik M, Szprengier-Juszkiewicz T, Jedziniak P (2010) Confirmatory method for determination of coccidiostats in eggs. Bull Vet Inst Pulawy 54:327–333

    Google Scholar 

  24. Cronly M, Behan P, Foley B, Malone E, Shearan P, Regan L (2010) Determination of eleven coccidiostats in animal feed by liquid chromatography–tandem mass spectrometry at cross contamination levels. Anal Chim Acta 700:26–33

    Article  Google Scholar 

  25. Boscher A, Guignard C, Pellet T, Hoffmann L, Bohn T (2010) Development of a multi-class method for the quantification of veterinary drug residues in feedingstuffs by liquid chromatography–tandem mass spectrometry. J Chromatogr A 1217:6394–6404

    Article  CAS  Google Scholar 

  26. Dubois M, Pierret G, Delahaut Ph (2004) Efficient and sensitive detection of residues of nine coccidiostats in egg and muscle by liquid chromatography–electrospray tandem mass spectrometry. J Chromatogr B 813:181–189

    Article  CAS  Google Scholar 

  27. Olejnik M, Szprengier-Juszkiewicz T, Jedziniak P (2009) Multi-residue confirmatory method for the determination of twelve coccidiostats in chicken liver using liquid chromatography tandem mass spectrometry. J Chromatogr A 1216:8141–8148

    Article  CAS  Google Scholar 

  28. Commission Decision 2002/657/EC (2002) Commission Decision of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off J Eur Communities L221:8–36

    Google Scholar 

  29. Beier RC, Stanker LH (1998) 4,4′-Dinitrocarbanilide—hapten development utilizing molecular models. Anal Chim Acta 376:139–143

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by Wessling International Research and Educational Center. We also wish to express our special thank for Kromat Ltd. for providing the LC–MS–MS instrument. We gratefully acknowledge Janssen for kindly donating diclazuril-bis and Merck Ltd. for providing the chemicals.

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Authors and Affiliations

  1. Joint Research and Training Laboratory on Separation Techniques, Eötvös Loránd University, Pázmány Péter sétány 1/A, Budapest, 1117, Hungary

    Szilárd Nász & Zsuzsanna Eke

  2. Alföldi Tej Kft, Seregélyesi út 127, Székesfehérvár, 8000, Hungary

    Etelka M. Károlyné

  3. Wessling International Research and Educational Center, Fóti út 56, Budapest, 1047, Hungary

    Tamás Rikker & Zsuzsanna Eke

Authors
  1. Szilárd Nász
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  2. Etelka M. Károlyné
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  3. Tamás Rikker
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  4. Zsuzsanna Eke
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Correspondence to Zsuzsanna Eke.

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Nász, S., Károlyné, E.M., Rikker, T. et al. Determination of Coccidiostats in Milk Products by LC–MS and Its Application to a Fermentation Experiment. Chromatographia 75, 645–653 (2012). https://doi.org/10.1007/s10337-012-2236-2

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  • DOI: https://doi.org/10.1007/s10337-012-2236-2

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