ANALYTICAL METHOD FOR DETERMINATION OF AVERMECTINS IN PORCINE LIVER BY LIQUID CHROMATOGRAPHY WITH FLUORESCENCE DETECTION
DOI:
https://doi.org/10.66211/tmvm.v11i2.341Keywords:
avermectins, HPLC-FLD, method validation, derivatization, liver, CLVCEAbstract
A method has been developed and validated for quantitative determination of residues of six avermectins - abamectin, doramectin, moxidectin, ivermectin, eprinomectin and emamectin in porcine liver by liquid chro-matography with fluorescence detection (HPLC-FLD). Nemadectin is used as an internal standard (IS). Aver-mectins are used as antiparasitic agents in veterinary medicine. They have maximum residue limits (MRLs) in accordance with Commission Regulation (EU) No 37/2010 in foodstuffs of animal origin. The sample prepa-ration is held with liquid extraction by acetonitrile, followed by clean up step using solid-phase extraction (SPE) with Strata C18-E cartridges and derivatization with trifluoroacetic acid anhydride (TFAA), 1-me-thylimidazole and the addition of 100 % acetic acid and triethylamine (TEA), to enable fluorescence detection. For the detection a column Agilent Eclipse XDB-C18 (150 x 4.6 mm, 5 μm) has been used, equipped with a guard column of the same packing and a fluorescence detector works with wavelengths at λex = 365 nm and λem = 460 nm. Gradient elution is applied with mobile phase A (acetonitrile) and mobile phase B (water) The analytical method is validated according to the requirements of Commission Implementing Regulation (EU) 2021/808. The method has been implemented and it is used as routine method for control of residues of Aver-mectins in Central Laboratory of Veterinary Control and Ecology (CLVCE).
References
1. European Commission, Commission Regulation (EU) No 37/2010 of 22 December 2009 on phar-macologically active substances and their classification regarding maximum residue limits in food-stuffs of animal origin, 2010. Available from: https://eur-lex.europa.eu/legal-con-tent/EN/TXT/?uri=CELEX%3A02010R0037-20240408 [Accessed 5 February 2026].
2. European Commission, Commission Implementing Regulation (EU) 2018/470 of 21 March 2018 on detailed rules on the maximum residue limit to be considered for control purposes for foodstuffs derived from animals which have been treated in the EU under Article 11 of Directive 2001/82/EC, 2018. Available from: https://eur-lex.europa.eu/eli/reg_impl/2018/470/oj [Accessed 5 February 2026].
3. EURL Guidance on minimum method performance requirements (MMPRs) for specific pharmaco-logically active substances in specific animal matrices. Version 3.0, 10.12.2025.
4. S. Sartori, et al. (2021). Classification, synthesis, biological activities and industrial applications, Tetrahedron. 84, 132001. doi: 10.1016/j.tet.2021.132001.
5. A. Crump, S. Ōmura. (2011). Ivermectin, 'wonder drug' from Japan: the human use perspective. Proc. Jpn. Acad. Ser. B. Phys. Biol. Sci., 87, 2, 13–28, https://doi.org/10.2183/pjab.87.13.
6. M. Danaher, et al. (2006). Review of methodology for the determination of macrocyclic lactone residues in biological matrices. J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 844, 2, 2006, 175–203. doi: 10.1016/j.jchromb.
7. Ruyu Sun, et al. (2023). Simultaneous Quantification of Avermectines in six Aquatic Foods by UHPLC/FLD with precolumn Derivatization. MDPI volume 10.
8. European Commission, Commission Implementing Regulation (EU) 2021/808 of 22 March 2021 on the performance of analytical methods for residues of pharmacologically active substances used in food-producing animals and on the interpretation of results as well as on the methods to be used for sampling and repealing Decisions 2002/657/EC and 98/179/EC, 2021. Available from: https://eur-lex.europa.eu/eli/reg_impl/2021/808/oj/eng [Accessed 5 February 2026]
9. M. Danaher, et al. (2001). Development and optimisation of an improved derivatisation procedure for determination of avermectins and milbemycins in bovine liver.
10. EURL Guidance Document on Stability. Studies in the Field of the Analysis of Residues of Phar-macologically Active Substances. Version 1.1, 28.01.2026
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 TRADITION AND MODERNITY IN VETERINARY MEDICINE
This work is licensed under a Creative Commons Attribution 4.0 International License.
