Epimers/Metabolites of Tetracycline Derivatives; Biological Activity and Regulation Aspects for MRL in Food

생물학적활성을 기초로 한 테트라싸이클린계 항생물질 잔류스크리닝법의 개선과 식품 중 잔류허용기준 설정 개선

  • Kwon, Jin-Wook (National Veterinary Research & Quarantine Service (NVRQS)) ;
  • Yun, Hyo-In (College of Veterinary Medicine, Chungnam Nat'l Univ.) ;
  • Lee, Kyu-Seung (Department of Applied Biology & Chemistry, College of Agriculture & Life Science, Chungnam Nat'l Univ.)
  • Received : 2011.02.23
  • Accepted : 2011.03.22
  • Published : 2011.03.31


BACKGROUND: Tetracyclines (TCs) are mainly regulated as parent compounds by bioactivity-based screening methods in food. Especially with respect to antimicrobial residues, their metabolites/epimers are also highly concerning chemicals and traditionally applied microbial detection methods are needed to improve with validation for regulatory control. METHODS AND RESULTS: Detection capability and biological activity of tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC) and their epimers; anhydrotetracycline (ATC), epianhydrotetracycline (EATC), epitetracycline (ETC), 4-epi-chlortetracycline (ECTC), 4-epianydrochlotetra-cycline (EACTC), 4-epioxychlortetracycline (EOTC), were measured by microbial growth inhibition screening method of Korea Food Code. CONCLUSION(S): Limited detection capabilities were found, B. megarerium and B. subtilis showed for TC and CTC, and B. subtilis for OTC. Biological potency of each epimer was also presented against various microorganisms, at the level from 50% to 96%, comparing with parent TCs. It is recommended that more advanced microbial screening methods with validation are needed, and biologically active epimers are to be considered as marker residues for MRL setting of regulatory control purpose.


  1. Aiello S.E., 1998. The Merck veterinary manual, 8th ed. Merck, Whitehouse Station, NJ, USA.
  2. Berendsen B.J.A., Pikkemaat, M. G., Stolker A.A.M., 2011. Are antibiotic screening approaches sufficiently adequate? A proficiency test, Analytica Chimica Acta 685;170-175.
  3. Bovee T.F., Pikkemaat M.G., 2009. Bioactivity-based screening of antibiotics and hormones. J. Chromatogr A. 13;1216(46):8035-50.
  4. Commission Decision 2002/657/EC, 2002. Implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off. J. Eur. Commun. L221/8.
  5. Community Reference Laboratories Residues (CRLs), 2010. Guidelines for the validation of screening methods for veterinary medicines (initial validation and transfer) 20/1/2010.
  6. Council Directive 96/23/EC, 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. Commun. L125.
  7. Duggar, B.M., 1948. A product of the continuing search for new antibiotics, Ann. N.Y. Acad.Sci. 51:177.
  8. EMEA, 1995. Committee for Veterinary Medicinal Products. Oxytetracycline, Tetracycline, Chlortetracycline. Summary Report (3). The European Agency for the Evaluation of Medicinal Products. EMEA/MRL/023/95.
  9. Food and Drug Administration, 2001. Guidance for Industry: Bioanalytical Method Validation. Rockville, MD: US Department of Health and Human Services, FDA, Center for Drug Evaluation and Research.
  10. Halling-Sorensen B., Sengelov G., Tjornelund J., 2002. Toxicity of tetracyclines and tetracycline degradation products to environmental relevant bacteria, including selected tetracycline-resistant bacteria. Arch. Environ. Contam. Toxicol. 42, 263–271.
  11. Okerman, L., De Wasch, K., van Hoof, J., Smedts, W., 2003. Simultaneous determination of different antibiotic residues in bovine and in porcine kidneys by solid-phase fluorescence immunoassay, J. AOAC. Int. 86, 236-240.
  12. Pikkemaat M.G., 2009. Microbial screening methods for detection of antibiotic residues in slaughter animals. Anal Bioanal Chem 395(4):893-905.
  13. Shah V.P., 2007. The history of bioanalytical method validation and regulation: Evolution of a guidance document on bioanalytical methods validation, The AAPS Journal 2007; 9 (1) Article 5.
  14. Soeborg T., Ingerslev F., Halling-Sorensen B., 2004. Chemical stability of chlortetracycline and chlortetracycline degradation products and epimers in soil interstitial water. Chemosphere 57(10):1515-24.