Control of Residues of Five Macrocyclic Lactones in Cow Milk By Liquid Chromatography with Fluorescence Detection

Teresa Szprengier-Juszkiewicz 1 , Piotr Jedziniak 1 , Małgorzata Olejnik 1  and Jan Żmudzki 1
  • 1 Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100 Pulawy, Poland

Abstract

A sensitive and reliable method has been developed and validated to determine residues of abamectin, doramectin, eprinomectin, ivermectin, and moxidectin in bovine milk. Isolation of the analytes from milk was performed with the use of liquidliquid extraction with acetonitrile in the presence of sodium chloride. The extract was defatted with hexane and cleaned up using solid phase extraction (C8 cartridge) after forming ion pairs with triethylamine. The analytes were derivatized with N, Ndimethylformamide, acetic acid anhydride, and N-methylimidazole (100°C, 90 min). The derivatives were determined by reverse phase liquid chromatography with fluorescence detection (excitation and emission wavelength 365 nm and 475 nm, respectively). Recoveries of the lactones from milk samples fortified at 10-30 μg kg-1 ranged from 52% to 80% with intra-laboratory reproducibility (CV) of 12.7%-22.8%. The critical concentrations (decision limit, CCα and detection capability, CCβ) were in accordance with target limits. The method has been verified in the proficiency studies by EURL/CVL Berlin (all z-scores in the range of ±2). The method was transferred to routine laboratories, verified in inter-laboratory comparison and successfully applied in the National Residue Control Plan.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • 1. Ali M.S., Sun T., McLeroy G.E., Phillippo E.T.: Simultaneous determination of eprinomectin, moxidectin, abamectin, doramectin and ivermectin in beef liver by LC with fluorescence detection. J AOAC Int 2000, 83, 31-38.

  • 2. Avermectins in milk (lyophilised samples). Interlaboratory Study AVER_07/05. Berlin 2006.

  • 3. Cerkvenik-Flajs V., Milčinski L., Süssinger A., Hodošček L., Danaher M., Antonić J.: Trace analysis of endectocides in milk by high performance liquid chromatography with fluorescence detection. Anal Chim Acta 2010, 663, 165-171.

  • 4. Commission Decision 2002/675/EC of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off J Eur Comm L 2002, 22, 8-36.

  • 5. Council Directive 96/23/EC of 29 April 1996 on measures to monitor certain substances and residues thereof in live animals and animal products and repealing Directives 85/358/EEC and 86/469/EEC and Decisions 89/187/EEC and 91/664/EEC. Off J Eur Comm L 1996, 125, 10-31.

  • 6. Council Regulation (EU) No 37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin Off J Eur Comm L 2010, 15, 1-72.

  • 7. Danaher M., Howells L.C., Crooks S.R.H., Cerkvenik- Flajs V., O’Keeffe M.: Review of methodology for the determination of macrocyclic lactone residues in biological matrices. J Chromatogr B 2006, 844, 175-203.

  • 8. Danaher M., O’Keeffe M., Glennon J.D.: Development and optimisation of an improved derivatisation procedure for the determination of avermectins and milbemycins in bovine liver. Analyst 2001, 126, 576-580.

  • 9. Danaher M., O’Keeffe M., Glennon J.D.: Validation and robustness testing of a HPLC method for the determination of avermectins and moxidectin in animal liver samples using an alumina column clean-up. Analyst 2000, 125, 1741-1744.

  • 10. Danaher M., Radeck W., Kolar L., Keegan J., Cerkvenik- Flajs V.: Recent developments in the analysis of avermectin and milbemycin residues in food safety and environment. Curr Pharm Biotechnol 2012, 13, 936-951.

  • 11. Deseleire E., Mortier L., Delahaut P.: Determination of macrocyclic lactones in bovine milk and meat tissue by liquid chromatography-tandem mass spectrometry. In: Proceedings of the EuroResidue V Conference on Residues of Veterinary Drugs in Food, edited by L.A.van Ginkel and A.A. Bergwerff, Noordwijkerhout, the Netherlands, 2004, pp. 422-428.

  • 12. Durden D.A.: Positive and negative electrospray LC-MSMS methods for quantitation of the antiparasitic endectocide drugs abamectin, doramectin, emamectin, eprinomectin, ivermectin, moxidectin and selamectin in milk. J Chromatogr B 2007, 850, 134-146.

  • 13. European Food Safety Authority; Report for 2010 on the results from the monitoring of veterinary medicinal product residues and other substances in live animals and animal products. Supporting Publications 2012:212. [65 pp.].

  • 14. FAO and WHO Codex Alimentarius. http://www.codexalimentarius.net/weg/uindex_en.jsp (accessed August 27, 2012)

  • 15. Lobato V., Rath S., Reyes F.G.R.: Occurrence of ivermectin in bovine milk from the Brazilian retail market. Food Addit Contam 2007, 23, 668-673.

  • 16. Nagata T.,F. Miyamoto Y., Hasegawa E.J.: Simultaneous determination of residual antiparasitic lactones in bovine muscle and liver by liquid chromatography with fluorescence detection. J AOAC Int 2003, 86, 490-493.

  • 17. Roudaut B.: Multiresidue method for the determination of avermectin and moxidectin residues in the liver using HPLC with fluorescence detection. Analyst 1998, 123, 2541-2544.

  • 18. Schenck F.J., Lagman L.H.: Multiresidue determination of abamectin, doramectin, ivermectin and moxidectin in milk using liquid chromatography and fluorescence detector. J AOAC Int 1999, 82, 1340-1344.

  • 19. Tolan J.W., Eskola P., Fink D.W., Mrozik H., Zimmerman L.A.: Determination of avermectins in plasma at nanogram levels using high-performance liquid chromatography with fluorescence detection. J Chromatogr 1980, 190, 367-376.

  • 20. Tway P.C., Wood J.S., Downing G.V.: Determination of ivermectin in cattle and sheep tissues using high performance liquid chromatography with fluorescence detection. J Agric Food Chem 1981, 29, 1059-1063.

  • 21. Żmudzki J., Niewiadowska A., Wojtoń B.: National veterinary residue control program in animal tissues and food of animal origin. Medycyna Wet 2005, 61, 649-653.

OPEN ACCESS

Journal + Issues

Search