Food Analytical Methods

, Volume 11, Issue 9, pp 2528–2537 | Cite as

Assessment of Oxytetracycline Residue in Cooked and Raw Meat of Chicken Broilers Before and After the End of Official Withdrawal Period

  • Kaddour Ziani
  • Marcos Pérez-López
  • Abdeldjallil Mansouri
  • Meghit Boumedienne Khaled
  • Antonio Silva Rodriguez
  • Miloud Slimani


The aim of this study was to determine and to compare the presence of oxytetracycline (OTC) residues in raw and cooked (boiled) meat before and after withdrawal period with the Maximum Residue Limits (MRL) using biological and physicochemical techniques. In animal building of 2000 broiler chickens (Gallus domesticus, breed ISA F15) and under the same conditions, 12 healthy adult chickens were selected and divided in two groups (n = 6 in each group). The sample was treated by Oxytetracycline® 20% (0.4 g/kg) during 3 days. Juices and muscles tissues were sampled twice for this assay: first, the broiler chickens were slaughtered (n = 6) 24 h after the third day of treatment. However, the second sampling (n = 6) was performed 24 h after the end of the withdrawal period. For each portions of sample, the juice meat and muscle tissues samples were collected after cooking at 80 °C during 45 min and after freezing/defrosting for the raw meat. Biological methods consisted of using four plates test (FPT) to pre-screening and Premi®Test to screening. Finally, to confirm and to quantify the accurate level of OTC in positive samples, ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) measurements were conducted. The FPT test showed that all samples were positive. While the Premi®Test analysis showed negative results for raw meat tested and positive results for juice of meat cooked in both samples. The obtained results using UHPLC-MS/MS methods confirmed the previously results. Thus, the presence of such toxic substances in our diet constitutes a major health risk for consumers, requiring the establishment of an adequate monitoring system. There is a paucity of official withdrawal periods for veterinary medicinal products in cooking meat; a relationship between the concentrations of residues of antibiotics after waiting period and cooking is not established since meat is always cooked before consumption.


Broiler chickens Cooking MRL OTC residues Withdrawal period 



The authors gratefully acknowledge to Dr. BENOUIS Bouziane for his technical assistance.

Compliance with Ethical Standards

Conflicts of Interest

Kaddour Ziani declares that he has no conflict of interest. Marcos Pérez-López declares that he has no conflict of interest. Mansouri Abdeldjallil declares that he has no conflict of interest. Meghit Boumedienne KHALED declares that he has no conflict of interest. Antonio Silva Rodriguez declares that he has no conflict of interest. Miloud Slimani declares that he has no conflict of interest.

Ethical Approval

In this article, all institutional and national guidelines for the care and use of laboratory animals were followed.

Informed Consent

Informed consent is not applicable for the nature of this study.


  1. Aarestrup F (2012) Get pigs off antibiotics. Nature 486:465–466CrossRefGoogle Scholar
  2. Aila O, Shitandi A, Mahungu SM, Sharma HK (2009) Determination of the depletion of furazolidone residues in chicken tissues using a Bacillus stearothermophilus test. J Food Control 20:543–547CrossRefGoogle Scholar
  3. Algerian Norm 2821 (1992) Fresh meat—research for antimicrobial residues. First edition pp. 1–3Google Scholar
  4. Al-Ghamdi M, Al-Mustafa Z, El-Morsy F, Al-Faky A, Haider I, Essa H (2000) Residues of tetracycline compounds in poultry products in the eastern province of Saudi Arabia. Public Health 114:300–304CrossRefGoogle Scholar
  5. Anadon A, Martinez-Larranaga MR (1999) Residues of antimicrobial drugs and feed additives in animal products: regulatory aspects. Livest Prod Sci 59:183–198CrossRefGoogle Scholar
  6. Anderson CR, Rupp HS, Wu WH (2005) Complexities in tetracycline analysis-chemistry, matrix extraction, cleanup, and liquid chromatography. J Chromatogr A 1075:23–32CrossRefGoogle Scholar
  7. Blasco C, Di Corcia A, Pico Y (2009) Determination of tetracyclines in multi-specie animal tissues by pressurized liquid extraction and liquid chromatography–tandem mass spectrometry. Food Chem 116:1005–1012CrossRefGoogle Scholar
  8. 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 feeding stuffs by liquid chromatography-tandem mass spectrometry. J Chromatogr A 1217:6394–6404CrossRefGoogle Scholar
  9. Cinquina AL, Longo F, Anastasi G, Giannetti L, Cozzani R (2003) Validation of a high-performance liquid chromatography method for the determination of oxytetracycline, tetracycline, chlortetracycline and doxycycline in bovine milk and muscle. J Chromatogr A 987:227–233CrossRefGoogle Scholar
  10. Codex Alimentarius Commission (2001) Committee on residues of veterinary drugs in foods, document control of veterinary drug residues in milk and milk products. Joint food and agriculture organization of the united nations world health organization food standards program 2001, 12th Ed. RomeGoogle Scholar
  11. Chafer-Pericas C, Maquieira A, Puchades R (2010) Fast screening methods to detect antibiotic residues in food samples. Trends Anal Chem 29:1038–1049CrossRefGoogle Scholar
  12. Cooper KM, Whelan M, Danaher M, Kennedy DG (2011) Stability during cooking of anthelmintic veterinary drug residues in beef. Food Addit Contam 28:155–165CrossRefGoogle Scholar
  13. Dang Pham K, Degand G, Douny C, Pierret G, Delahaut P, Ton VD, Granier B, Scippo M-L (2013) Preliminary evaluation of antimicrobial residue levels in marketed pork and chicken meat in the Red River Delta region of Vietnam. Food Public Health 3:267–276Google Scholar
  14. De Ruyck H, De Ridder H (2007) Determination of tetracycline antibiotics in cow’s milk by liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 21:1511–1520CrossRefGoogle Scholar
  15. European Commission, Commission Regulation 37/2010(2010) Pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin. Off J Eur Commun L15 (1)Google Scholar
  16. Franco DA, Webb J, Taylor CE (1990) Antibiotic and sulfonamide residues in meat: implications for human health. J Food Prot 53:178–185CrossRefGoogle Scholar
  17. Gaudin V, Juhel-Gaugain M, Moretain J, Sanders P (2008) AFNOR validation of Premi®test, a microbiological-based screening tube-test for the detection of antimicrobial residues in animal muscle tissue. Food Addit Contam 25:1451–1464CrossRefGoogle Scholar
  18. Gaudin V, Hedou C, Rault A, Sanders P, Verdon E (2009) Comparative study of three screening tests, two microbiological tube tests, and a multi-sulphonamide ELISA kit for the detection of antimicrobial and sulphonamide residues in eggs. Food Addit Contam 26:427–440CrossRefGoogle Scholar
  19. Gaudin V, Celine H, Annie R, Eric V (2010) Validation of a five plate test, the STAR protocol, for the screening of antibiotic residues in muscle from different animal species according to the European decision 2002/657/EC. Food Addit Contam 27:935–952CrossRefGoogle Scholar
  20. Gratacós-Cubarsí M, Fernandez-García A, Picouet P, Valero-Pamplona A, García-Regueiro JA, Castellari M (2007) Formation of tetracycline degradation products in chicken and pig meat under different thermal processing conditions. J Agric Food Chem 55:4610–4616CrossRefGoogle Scholar
  21. Hsieh MK, Shyu CL, Liao JW, Franje CA, Huang YJ, Chang SK, Chou CC (2011) Correlation analysis of heat stability of veterinary antibiotics by structural degradation, changes in antimicrobial activity and genotoxicity. Veterinarni Medicina 56:274–285CrossRefGoogle Scholar
  22. Javadi A (2011) Effect of roasting, boiling and microwaving cooking method on doxycline residues in edible tissues of poultry by microbial method. Afr J Pharm Pharmacol 5:1034–1037Google Scholar
  23. Koesukwiwat U, Jayanta S, Leepipatpiboon N (2007) Validation of a liquid chromatographye mass spectrometry multi-residue method for the simultaneous determination of sulfonamides, tetracyclines, and pyrimethamine in milk. J Chromatogr A 1140:147–156CrossRefGoogle Scholar
  24. Tian L, Khalil S, Bayen S (2016) Effect of thermal treatments on the degradation of antibiotic residues in food. Crit Rev Food Sci Nutr 57:3760–3770CrossRefGoogle Scholar
  25. Lolo M, Pedreira S, Miranda JM, Vázquez BI, Franco CM, Cepeda A, Fente C (2006) Effect of cooking on enrofloxacin residues in chicken tissue. Food Addit Contam 23:988–993CrossRefGoogle Scholar
  26. Ministry of Agriculture and Rural Development (MARD) (2016) Agricultural poles. Available at Accessed November 20, 2016
  27. Mourot D, Loussourorn S (1981) Sensitivity of lactic acid bacteria to antibiotics used in veterinary medicine (in French). Rec Med Vet Ec Alfort 157:175–177Google Scholar
  28. Myllyniemi AL, Rintala R, Backman C, Niemi A (1999) Microbiological and chemical identification of antimicrobial drugs in kidney and muscle samples of bovine cattle and pigs. Food Addit Contam 16:339–351CrossRefGoogle Scholar
  29. Myllyniemi AL, Sipila H, Nuotio L, Niemi AA, Honkanen BT (2002) An indirect conductimetric screening method for the detection of antibiotic residues in bovine kidneys. Analyst 127:1247–1251CrossRefGoogle Scholar
  30. Nguyen V, Li M, Khan M, Li C, Zhou G (2013) Effect of cooking methods on tetracycline residues in pig meat. African J Pharm Pharmacol 7:1448–1454CrossRefGoogle Scholar
  31. Nguyen V, Nguyen V, Li C, Zhou G (2015) The degradation of oxytetracycline during thermal treatments of chicken and pig meat and the toxic effects of degradation products of oxytetracycline on rats. J Food Sci Technol 52:2842–2850CrossRefGoogle Scholar
  32. Nina E, Virolainen Mariël G, Pikkemaat JW, Elferink A, Karp MT (2008) Rapid detection of Tetracyclines and their 4-Epimer derivatives from poultry meat with bioluminescent biosensor bacteria. J Agric Food Chem 56:11065–11070CrossRefGoogle Scholar
  33. Okerman L, Croubels S, De Baere S, Hoof JV, De Backer P, De Brabander H (2001) Inhibition tests for detection and presumptive identification of tetracyclines, beta-lactam antibiotics and quinolones in poultry meat. Food Addit Contam 18:385–393CrossRefGoogle Scholar
  34. Pikkemaat MG, Rapallini MLBA, Oostra-Van Dijk S, Elferink JWA (2009) Comparison of three microbial screening methods for antibiotics using routine monitoring samples. Anal Chim Acta 637:298–304CrossRefGoogle Scholar
  35. Pikkemaat MG, Rapallini MLBA, Zuidema T, Elferink JWA, Oostra-Van Dijk S (2010) Screening methods for detection of antibiotic residues in slaughter animals: comparison of the EU-four plate method, the Nouws Antibiotic Test and the Premi®Test (applied to muscle and kidney). Food Addit Contam 28:1–26Google Scholar
  36. Rose MD, Bygrave J, Farrington WH, Shearer G (1996) The effect of cooking on veterinary drug residues in food: 4. Oxytetracycline. Food Addit Contam 13:275–286CrossRefGoogle Scholar
  37. Shalaby AR, Nadia AS, Abou-Raya SH, Wafaa HE, Mehaya FM (2011) Validation of HPLC method for determination of tetracycline residues in chicken meat and liver. Food Chem 124:1660–1666CrossRefGoogle Scholar
  38. Sierra D, Sánchez A, Contreras A, Luengo C, Corrales JC, Morales CT, de la Fe C, Guirao, Gonzalo C (2009) Detection limits of four antimicrobial residue screening tests for beta-lactams in goat’s milk. J Dairy Sci 92:3585–3591CrossRefGoogle Scholar
  39. Traub WH, Leonhard B (1995) Heat stability of the antimicrobial activity of sixty-two antibacterial agents. J Antimicrob Chemother 35:149–154CrossRefGoogle Scholar
  40. USFDA ERA (1989) Environmental assessment Oxytet soluble (oxytetracycline hydrochloride soluble powder). I.D. Russell Company, Laboratories 368–379Google Scholar
  41. Van Boeckel TP, Brower C, Gilbert M, Grenfell BT, Levin SA, Robinsoni TP (2015) Global trends in antimicrobial use in food animals. Proc Natl Acad Sci 112:5649–5654CrossRefGoogle Scholar
  42. Wen Y, Wang Y, Feng Y (2006) Simultaneous residue monitoring of four tetracycline antibiotics in fish muscle by in-tube solid-phase microextraction coupled with high-performance liquid chromatography. Talanta 70:153–159CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory of Biotoxicology, Pharmacognosy and Biological Valorization of Plants, Department of BiologyTaher Moulay University of SaidaSaidaAlgeria
  2. 2.Toxicology AreaFaculty of Veterinary Medicine (UEX)CaceresSpain
  3. 3.Health and Environment laboratoryDjillali Liabes University of Sidi-Bel-AbbesSidi-Bel-AbbesAlgeria
  4. 4.Animal Source Foodstuffs Innovation and Analysis Service (SiPA)University of ExtremaduraBadajozSpain

Personalised recommendations