THE DEVELOPMENT AND THE VALIDATION OF THE METHOD FOR TRIMETOPRIM RESIDUES DETERMINATION IN TISSUES SAMPLES USING THE METHOD OF ENZYME-LINKED IMMUNOSORBENT ASSAY
The article presents the results of the development of sample preparation method of muscle and parenchymal tissues for their further testing on trimethoprim residues by enzyme-linked immunosorbent assay using Trimethoprim test kits from Kwinbon Biotechnology (China), designed to analyze another target matrix, viz. honey. The results of the comparison of sample preparation methods with the use of different techniques of analyte extraction from tissues, with the established percentage of extraction are presented. The most effective conditions are proposed: acetonitrile extraction using ultrasonic intensification, subsequent concentration of the analyte by evaporation, reconstitution of dried residue and degreasing. This approach makes it possible to determine trimethoprim residues in poultry tissues at the level of 5-10 μg/kg with an absolute recovery of about 75%. In accordance with the Decision of the European Commission 2002/657/EC, the validation of the proposed method was carried out taking into account the maximum permissible levels (MRL) of trimethoprim in the tissues of productive animals. The suitability of the method was confirmed on the basis of establishing of the main validation parameters for screening methods (technical threshold and cut-off factor) using control (pure) and spiked with a standard solution of trimethoprim at the level of ½ MRL samples of muscle and parenchymal tissues of broiler chickens. The main advantages of the developed technique are simplicity of performance, rapidity and cost effectiveness. The certainty of the results obtained using the developed method was verified using the confirmatory UPLC-MS/MS method. The article presents the results of a comparative study of pure samples of broiler chickens muscles and parenchymal tissues and spiked at the level of ½ MRL with a standard solution of trimethoprim. The developed screening technique of enzyme-linked immunosorbent assay is recommended for determination of residual amounts of trimethoprim in tissue samples and can be used for routine laboratory tests, as well as to study the withdrawal procedure of veterinary drugs based on this drug.
Abd Elkhabeer, M., Ramadan, G.A., Ryad Eglal, L., Souaya, R. (2020). Analytical Method Optimization and Determination of Sulfonamides, Chloramphenicol and Tetracyclines Drug Residues in Chicken Meat across Egypt. Journal of Food Processing & Technology. 11(4)
Amini, H. & Ahmadiani, A. (2007). Rapid and simultaneous determination of sulfamethoxazole and trimethoprim in human plasma by high-performance liquid chromatography. Journal of Pharmaceutical and Biomedical Analysis. 43(3). 1146-1150 DOI.org/10.1016/j.jpba.2006.09.004
Andrée, S. & Schwägele, F. (2007). Determination of veterinary drug residues in poultry meat – Sulfonamides and trimethoprim. Fleischwirtschaft –Frankfurt. 87(6). 104-108.
Baert, K, De Baere, S., Croubels, S., De Backer. P. (2003). Pharmacokinetics and oral bioavailability of sulfadiazine and trimethoprim in broiler chickens. Veterinary Research Communications. 27(4), 301-309. DOI: 10.1023/a:1024084108803
Biswas, A.K., Rao, G.S., Kondaiah, N., Anjaneyulu. A.S.R., Malik, J.K. (2007). Simple Multiresidue Method for Monitoring of Trimethoprim and Sulfonamide Residues in Buffalo Meat by High-Performance Liquid Chromatography. Journal of Agriculture and Food Chemistry. 55(22), 8845-8850. DOI: org/10.1021/jf071140w
Bousova, K.. & Mittendorf, K. (2016). Automated Online Multi-Residue LC-MS/MS Method for the Determination of Antibiotics in Chicken Meat. Method number: 63646. Thermo Fisher Scientific.
Brands, E., Kubalec, P., Machác, L. (1997). HPLC determination of trimethoprim in meat and milk with an SPE preseparation procedure. Zeitschrift für Lebensmitteluntersuchung und Forschung A. 204. 341-344.
CODEX ALIMENTARIUS (2012). Maximum residue limits for veterinary drugs in foods updated as at the 35th Session of the Codex Alimentarius Commission, CAC/LMR 2.
De Baere, S., Baert, K., Croubels, S., De Busser, J., De Wasch, K., De Backer. P. (2000). Determination and quantification of sulfadiazine and trimethoprim in swine tissues using liquid chromatography with ultraviolet and mass spectrometric detection. Analyst. 125. 409-415. DOI: 10.1039/a908750h
European Commission (2002). Commission Decision 2002/657/EC of 14 august 2002 on implementing Council directive 96/23/EC concerning the performance of analytical methods and the interpretation of results 2002/657/EC. Official Journal of European Community. L. 221. 8-36.
European Council (2010). Council Regulation 37/2010/EU of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin. Official Journal of European Union. L15. 1-72.
Hui, Li, Hanwen, S., Jingxuan, Zh., Kun, P. (2013). Highly sensitive and simultaneous determination of sixteen sulphonamide antibiotics, four acetyled metabolites and trimethoprim in meat by rapid resolution liquid chromatography-tandem mass spectrometry. Food Control. 31(2). 359-365.
Mohamed, D. & Fouad, M.M. (2020). Application of NEMI, Analytical Eco-Scale and GAPI tools for greenness assessment of three developed chromatographic methods for quantification of sulfadiazine and trimethoprim in bovine meat and chicken muscles: Comparison to greenness profile of reported HPLC methods. Microchemical Journal. 157. 104873
Pfeifer, T., Tuerk, J., Bester, K., Spiteller, M. (2002). Determination of selected sulfonamide antibiotics and trimethoprim in manure by electrospray and atmospheric pressure chemical ionization tandem mass spectrometry. Rapid Communications in Mass Spectrometry. 6(7). 663-669. DOI: 10.1002/rcm.624.
Pikkemaat, M.G., Rapallini, M.L.B.A., Zuidema, T., Elferink, J.W.A., Oostra-van Dijk, S., Driessen-van Lankveld, W.D.M. (2011). Screening methods for detection of antibiotic residues in slaughter animals: comparison of the EU-four plate method, the Nouws Antibiotic Test and the Premi®Tedt (applied to muscle and kidney). Food Additives & Contaminants: Part A:
Chemistry, Analysis, Control, Exposure & Risk Assessment. 28(1). 26-34. DOI: 10.1080/19440049.2010.535027.
Sayar, E., Sahin, S., Cevheroglu, S., Atilla Hincal, A. (2010). Development and validation of an HPLC method for simultaneous determination of trimethoprim and sulfamethoxazole in human plasma. European Journal of Drug Metabolism and Pharmacokinetics. 35. 41-46.
Soto-Chinchilla, J.J, García-Campaña, A.M, Gamiz-Gracia, L. (2007). Analytical methods for multiresidue determination of sulfonamides and trimethoprim in meat and ground water samples by CE-MS and CE-MS/MS. Electrophoresis. 28(22). 4164-4172. DOI: 10.1002/elps.200600848.
Varenina, I., Bilandzic, N., Kolanovic, B.S., Bozic, D., Sedak, M., Dokic, M. (2016). Validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of sulfonamides, trimethoprim and dapsone in muscle, egg, milk and honey. Food Additives & Contaminants: Part A: Chemistry, Analysis, Control, Exposure & Risk Assessment. 33(4). 656-667. DOI.org/10.1080/19440049.2016.1152569
Wang, S., Zhang, H.Y., Wang, L., Duan, Z.J., Kennedy, I. (2006). Analysis of sulphonamide residues in edible animal products: A review. Food Additives & Contaminants. 23(4), 362-384.
Yanovych. D., Zasadna, Z., Rydchuk, M., Plotycya, S., Kislova, S., Pazderska, O. (2020). The development of the method for determination of sulfonamides and trimethoprim residual quantities in milk samples by UPLC-MS/MS method and its verification through interlaboratory testing. 21(2) 170-182. DOI: 10.36359/scivp.2020-21-2.23.
This work is licensed under a Creative Commons Attribution 4.0 International License.