The study was conducted with the aim of developing and validating a method for determining the content of some tetracycline group substances including tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC), metacycline (MC), doxycycline (DC) and epimers: 4-epi oxytetracycline (4-epi OTC), 4-epi tetracycline (4-epi TC), 4-epi chlortetracycline (4-epi CTC) in meat and meat products by liquid chromatography tandem mass spectrometry (LC-MS/MS). The sample processing procedure was quick and simple: the sample was extracted with McIlvaine-EDTA buffer, and the protein was precipitated with trichloroacetic acid. Then, the extract was cleaned on a HLB solid-phase extraction column. The separation was performed on a reversed-phase X-bridge C18 column (150 mm x 2.1 mm id; 3.5 μm), mobile phase channel A was 0.1% formic acid in H2O, channel B was ACN; flow rate 0.5 mL/min, injection volume 10 μL. The validated method showed good specificity, the standard curve of the analytes was linear in the concentration range of 15 – 150 μg/kg. The limit of detection (LOD) and limit of quantification (LOQ) were achieved with the corresponding analytes of 5 μg/kg and 15 μg/kg. The accuracy was assessed by recovery in the range of 84.23 – 108.5%, the precision was assessed by repeatability and internal reproducibility met the requirements according to AOAC. The method was applied to analyze the content of tetracycline antibiotics and epimers in 10 food samples (chicken, pork, ham, sausage) collected from the market.
TC, OTC, CTC, 4-epi OTC, 4-epi TC, 4-epi CTC, meat, meat products, LC/MS-MS
[1]. G. Evtugyn, A. Porfireva, G. Tsekenis, V. Oravczova, and T. J. S. Hianik, "Electrochemical aptasensors for antibiotics detection: Recent achievements and applications for monitoring food safety," Sensors (Basel), vol. 22, no. 10, p. 3684, 2022. doi: 10.3390/s22103684.
[2]. I. Chopra, M. J. M. Roberts, and m. b. reviews, "Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance," Microbiology and Molecular Biology Reviews, vol. 65, no. 2, pp. 232-260, 2001. doi: 10.1128/MMBR.65.2.232-260.2001.
[3]. Y. Amangelsin, Y. Semenova, M. Dadar, M. Aljofan, and G. Bjørklund, "The Impact of Tetracycline Pollution on the Aquatic Environment and Removal Strategies," Antibiotics (Basel, Switzerland), vol. 12, no. 3, 2023. doi: 10.3390/antibiotics12030440.
[4]. M. G. Bacanlı, "The two faces of antibiotics: an overview of the effects of antibiotic residues in foodstuffs," Archives of toxicology, vol. 98, no. 6, pp. 1717-1725, Jun 2024, doi: 10.1007/s00204-024-03760-z.
[5]. Ministry of Health, "24/2013/TT-BYT, Maximum Limits On Residues Of Veterinary Medicine In Food," 2013 [Online]. Available https://thuvienphapluat.vn/van-ban/Bo-may-hanh-chinh/Thong-tu-24-2013-TT-BYT-muc-gioi-han-toi-da-du-luong-thuoc-thu-y-204380.aspx.
[6]. Mai Hoang Ngoc Do et al., "Screening of antibiotic residues in pork meat in Ho Chi Minh City, Vietnam, using a microbiological test kit and liquid chromatography/tandem mass spectrometry," Food control, vol. 69, pp. 262-266, 2016, doi: 10.1016/j.foocont.2016.05.004.
[7]. European Commission, "Commission regulation (EC) 1442/95, 1995, of 26 June 1995 amending annexes I, II, III and IV to regulation (EEC) no. 2377/90 laying down a community procedure for the establishment of maximum residue limits of veterinary medicinal products in foodstuffs of animal origin.," 1995.
[8]. TCVN 8748:2011, "Meat and meat products - Method for determination of tetracyclines residues by high performance liquid chromatography," 2011 (in Vietnamese). [Online]. Available https://luatvietnam.vn/thuc-pham/tieu-chuan-tcvn-8748-2011-xac-dinh-du-luong-tetracyclin-trong-thit-161794-d3.html.
[9]. S. Kruanetr, K. J. C. A. S. Prasertboonyai, and TECHNOLOGY, "UV-Vis Spectrophotometric Method Using Natural Reagent from Vigna unguiculata subsp. sesquipedalis for Tetracycline Determination in Pharmaceutical Samples," Current Applied Science and Technology, vol. 21, no. 1, 2021.
[10]. Y. A. Akeneev, G. B. Slepchenko, V. I. Deryabina, T. M. Gindullina, I. O. Maximchuk, and T. I. Shchukina, "Determination of Tetracycline in Honey by Voltammetry," Procedia Chemistry, vol. 15, pp. 355-359, 12/31 2015, doi: 10.1016/j.proche.2015.10.056.
[11]. N. T. Huong, N. T. M. Hien, T. T. D. Thuy, N. T. N. Ai, and L. D. Q. J. V. J. o. C. Anh, "Determination of tetracycline residue in pork by differential pulse voltammetry using activated glassy carbon electrode," Vietnam journal of chemistry, vol. 61, no. 1, pp. 65-73, 2023, doi: 10.1002.vjch.202200010.
[12]. F. Alanazi, R. Almugbel, H. M. Maher, F. M. Alodaib, and N. Z. Alzoman, "Determination of tetracycline, oxytetracycline and chlortetracycline residues in seafood products of Saudi Arabia using high performance liquid chromatography-Photo diode array detection," Saudi pharmaceutical journal : SPJ : the official publication of the Saudi Pharmaceutical Society, vol. 29, no. 6, pp. 566-575, Jun 2021, doi: 10.1016/j.jsps.2021.04.017.
[13]. E. Patyra and K. J. J. o. V. R. Kwiatek, "Analytical procedure for the determination of tetracyclines in medicated feedingstuffs by liquid chromatography-mass spectrometry," Journal of Veterinary Research, vol. 60, no. 1, pp. 35-41, 2016, doi: 10.1515/jvetres-2016-0006.
[14]. N. T. T. Hang and N. M. J. V. J. o. F. C. Khai, "Determination of macroline and tetracyclines group in water by liquid chromatography tandem mass spectrometry (LC-MS/MS)," Vietnam Journal of Food Control, vol. 3, no. 3, pp. 173-182, 2020, doi: 10.47866/2615-9252/vifc.1723.
[15]. A. Jia, Y. Xiao, J. Hu, M. Asami, and S. J. J. o. C. A. Kunikane, "Simultaneous determination of tetracyclines and their degradation products in environmental waters by liquid chromatography–electrospray tandem mass spectrometry," Journal of Chromatography A, vol. 1216, no. 22, pp. 4655-4662, 2009, doi: 10.1016/j.chroma.2009.03.073.
[16]. A. Gajda and A. J. J. o. V. R. Posyniak, "Liquid chromatography–tandem mass spectrometry method for the determination of ten tetracycline residues in muscle samples," Bulletin of the Veterinary Institute in Pulawy, vol. 59, no. 3, pp. 345-352, 2015.
[17]. D. A. Bohm, C. S. Stachel, P. J. A. Gowik, and B. Chemistry, "Validation of a multi-residue method for the determination of several antibiotic groups in honey by LC-MS/MS," Analytical and Bioanalytical Chemistry, vol. 403, pp. 2943-2953, 2012.
[18]. J.-Z. Xu et al., "Analysis of tetracycline residues in royal jelly by liquid chromatography–tandem mass spectrometry," Journal of Chromatography B, vol. 868, no. 1, pp. 42-48, 2008/06/01/ 2008, doi: 10.1016/j.jchromb.2008.04.027.
[19]. C. R. Anderson, H. S. Rupp, and W.-H. J. J. o. C. A. Wu, "Complexities in tetracycline analysis—chemistry, matrix extraction, cleanup, and liquid chromatography," Journal of Chromatography A, vol. 1075, no. 1-2, pp. 23-32, 2005, doi: 10.1016/j.chroma.2005.04.013.
[20]. D. Rajalingam, C. Loftis, J. J. Xu, and T. K. Kumar, "Trichloroacetic acid-induced protein precipitation involves the reversible association of a stable partially structured intermediate," Protein science: a publication of the Protein Society, vol. 18, no. 5, pp. 980-93, May 2009, doi: 10.1002/pro.108.
[21]. G. Latimer Jr, "Guidelines for standard method performance requirements: official methods of analysis," AOAC International, pp. 1-18, 2016.
[22]. F. Tasci, H. S. Canbay, and M. J. F. C. Doganturk, "Determination of antibiotics and their metabolites in milk by liquid chromatography-tandem mass spectrometry method," Food Control, vol. 127, p. 108147, 2021, doi: 10.1016/j.foodcont.2021.108147.