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Improvement of an Simultaneous Determination for Clenbuterol and Ractopamine in Livestock Products using LC-MS/MS

LC-MS/MS를 이용한 축산물 중 clenbuterol과 ractopamine의 동시 분석법 개선

  • Received : 2012.11.29
  • Accepted : 2013.01.21
  • Published : 2013.02.28

Abstract

Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

Keywords

clenbuterol;ractopamine;beef;pork;LC-MS/MS

References

  1. FAO/WHO-JECFA. Evaluation of certain veterinary drug residues in food: Seventy-fifth report of the joint FAO/WHO expert committee on food additives (WHO technical report series; No. 969). FAO/WHO, Geneva, Switzerland. pp. 1-101 (2012)
  2. MHLW. Available from: http://www.mhlw.go.jp/english/topics/mrls/final/mrls.html. Accessed May 31, 2005.
  3. FDA. Available from: http://www.fda.gov/Food/FoodSafety/default.htm. Accessed January 30, 2013.
  4. European Union. Commission Regulation (EU) No. 37/2010: On pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin. Off. J. Eur. Union L 15: 1-72 (2010)
  5. Oh JH, Kwon CH, Jeon JS, Choi DM. Management of veterinary drug residues in food. Korean J. Environ. Agric. 28: 310-325 (2009) https://doi.org/10.5338/KJEA.2009.28.3.310
  6. European Union. Council 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. Off. J. Eur. Union L 224: 1-8 (1990)
  7. KFDA. Notice No. 2007-68. Available from: http://kfda.go.kr/ index.kfda?mid=95. Accessed October 18, 2007.
  8. KFDA. Notice No. 2012-17. Available from: http://kfda.go.kr/ index.kfda?mid=92. Accessed May 10, 2012.
  9. Park MS, Lee JJ, Myung SW. Analysis of phoxim residue in animal food production (cattle and pig) by LC/ESI-MS/MS. J. Korean Chem. Soc. 55: 626-632 (2011) https://doi.org/10.5012/jkcs.2011.55.4.626
  10. KFDA. 2011 Safety evaluation on residues of veterinary drugs in foods. Korea Food and Drug Administration, Cheongwon, Korea (2011)
  11. KFDA. Food code. Korea Food and Drug Administration, Cheongwon, Korea. pp. 10.5.121-123 (2011)
  12. KFDA. Food code. Korea Food and Drug Administration, Cheongwon, Korea. pp. 10.5.161-164 (2011)
  13. Juan C, Igualada C, Moragues F, Leon N, Manes J. Development and validation of a liquid chromatography tandem mass spectrometry method for the analysis of $\beta$-agonists in animal feed and drinking water. J. Chromatogr. A 1217: 6061-6068 (2010) https://doi.org/10.1016/j.chroma.2010.07.034
  14. KFDA. Clenbuterol risk profile. Available from: http://www.foodnara. go.kr/foodnara/board-list.do Accessed December, 2010.
  15. Elanco. Material safety data sheet. Available from: http://www.elancocentral.com/topmax_msds.pdf. Accessed October 17, 2008.
  16. Blanca J, Muñoz P, Morgado M, Méndez N, Aranda A, Reuvers T, Hooghuis H. Determination of clenbuterol, ractopamine and zilpaterol in liver and urine by liquid chromatography tandem mass spectrometry. Anal. Chim. Acta 529: 199-205 (2005) https://doi.org/10.1016/j.aca.2004.09.061
  17. Shishani E(I), Chai SC, Jamokha S, Aznar G, Hoffman MK. Determination of ractopamine in animal tissues by liquid chromatography- fluorescence and liquid chromatography/tandem mass spectrometry. Anal. Chim. Acta 483: 137-145 (2003) https://doi.org/10.1016/S0003-2670(03)00120-X
  18. Lee DW, Mona IC, Daniel RD. Multiresidue confirmation of $\beta$- agonists in bovine retina and liver using LC-ES/MS/MS. J. Chromatogr. B 813: 35-45 (2004) https://doi.org/10.1016/j.jchromb.2004.09.005
  19. Li C, Wu YL, Yang T, Zhang Y, Huang-Fu WG. Simultaneous determination of clenbuterol, salbutamol, and ractopamine in milk by reversed-phase liquid chromatography tandem mass spectrometry with isotope dilution. J. Chromatogr. A 1217: 7873-7877 (2010) https://doi.org/10.1016/j.chroma.2010.10.055
  20. Fesser ACE, Dickson LC, Macneil JD, Patterson JR, Lee S, Gedir RD. Determination of $\beta$-agonists in liver and retina by liquid chromatography-tandem mass spectrometry. J. AOAC Int. 88: 61-69 (2005)
  21. Widstrand C, Larsson F, Fiori M, Civitareale C, Mirante S, Brambilla G. Evaluation of MISPE for the multi-residue extraction of $\beta$-agonists from calves urine. J. Chromatogr. B 804: 85-91 (2004) https://doi.org/10.1016/j.jchromb.2003.12.034
  22. Xu Z, Hu Y, Hu Y, Li G. Investigation of ractopamine molecularly imprinted stir bar sorptive extraction and its application for trace analysis $\beta_{2}$-agonists in complex samples. J. Chromatogr. A 1217: 3612-3618 (2010) https://doi.org/10.1016/j.chroma.2010.03.046
  23. Masci G, Casati G, Crescenzi V. Synthesis and LC characterization of clenbuterol molecularly imprinted polymers. J. Pharmaceut. Biomed. 25: 211-217 (2001) https://doi.org/10.1016/S0731-7085(00)00477-5
  24. Codex Alimentarius Committee. Codex guidelines for the estab lishment of a regulatory programme for control of veterinary drug residues in foods, CAC/GL 16. Codex, Rome, Italy. pp. 1-46 (1993)
  25. Zhang X, Chen L, Xu Y, Wang H, Zhao Q, Zhao Q, Ren N, Ding L. Determination of $\beta$-lactam antibiotics in milk based on magnetic molecularlyimprinted polymer extraction coupled with liquid chromatography-tandem mass spectrometry. J. Chromatogr. B 878: 3421-3426 (2010) https://doi.org/10.1016/j.jchromb.2010.10.030
  26. Guy PA, Savoy MC, Stadler RH. Quantitative analysis of clenbuterol in meat products using liquid chromatography-electrospray ionisation tandem mass spectrometry. J. Chromatogr. B 736: 209- 219 (1999) https://doi.org/10.1016/S0378-4347(99)00466-1
  27. EU Council. Implementing council directive 96/23/EC concerning the performance of analytical methods and the interpretation of results (2002/657/EC). Off. J. Eur. Commun. L221: 8-36 (2002)

Cited by

  1. Establishment of a Method for Analyzing the Zilpaterol Residue in Beef Using Tandem Mass Spectrometry vol.20, pp.6, 2014, https://doi.org/10.3136/fstr.20.1165

Acknowledgement

Grant : 2011년 잔류동물의 약품 시험법 개발 및 잔류실태조사

Supported by : 식품의약품안전평가원