Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Determination of Four Macrolide Antibiotics Residues in Chicken Muscle Using High-Performance Liquid Chromatography

액체크로마토그래피를 이용한 닭고기 시료에서의 마크로라이드계 동시분석법 개발

  • 이상희 (한국식품연구원, 식품분석센터) ;
  • 유미영 (한국식품연구원, 식품분석센터) ;
  • 신동빈 (한국식품연구원, 식품분석센터)
  • Received : 2012.09.05
  • Accepted : 2012.12.24
  • Published : 2013.03.31
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Abstract

A simple and rapid method has been developed and validated for simultaneous determination of each macrolides residues (spiramycin, josamycin, tilmicosin, tylosin) in chicken muscle by high-performance liquid chromatography- photo diode array (HPLC-PDA). Chicken muscle sample have been extracted with liquid-liquid extraction process; analytes was extracted by acetonitrile, and then defatted with hexane saturated by acetonitrile. The HPLC separation was performed on a Unison UK-$C_{18}$ ($150mm{\times}3.0mm$, $3{\mu}m$) with a gradient system of 0.1% trifloroacetic acid (TFA) and 0.1% trifloroacetic acid (TFA) in acetonitrile as the mobile phase. The drugs were detected at 232 nm for spiramycin and josamycin, and 287 nm for tilmicosin and tylosin. The limits of quantification (LOQs) were between 27 and $59{\mu}g/kg$; and the intra- and inter-day precision (relative standard deviation; RSD) was between 0.9-13.2 and 2.4-13.1%, respectively in chicken muscle sample. The method may has been successfully applied for multiresidue determination of four macrolides below the maximum residue limits (MRLs) established by the European Union (EU).

본 논문에서는 축산물 중 닭고기 내 마크로라이드계(MLs) 항생물질(spiramycin, josamycin, tilmicosin, 그리고 tylosin) 4종을 분석하기 위하여, 액-액 추출 과정을 거쳐서 PDA 검출기가 장착된 액체크로마토그래피를 이용하여 MLs를 효율적으로 동시분석하는 방법을 확립하였다. 컬럼은 Unison UK-C18 ($150mm{\times}3mm$ id, $3{\mu}m$), 이동상 용매는 0.1% TFA와 0.1% TFA in ACN로 기울기 용리를 사용하였으며, 유속은 0.7 mL/min, 그리고, 주입량은 $10{\mu}L$로 설정하여 분석하였다. 확립된 분석조건으로, 표준검정 곡선은 $50-1000{\mu}g/kg$의 농도범위에서 상관계수가 0.9975 이상의 양호한 직선성을 나타내었으며, 회수율은 저, 중, 고농도로 첨가하여 분석한 결과, 80.4-99.1%를 나타내었다. 검출한계는 $8.8-19.6{\mu}g/kg$이었고, 정량한계는 $26.6-59.4{\mu}g/kg$이었으며, 일내(intra-day)와 일간(inter-day) 정밀도(CV%)는 0.9-13.2%, 2.4-13.1%이었다. 따라서, 확립된 분석방법은 축산물 중 닭고기 시료 내 MLs를 효과적으로 분석하는데 이용될 수 있을 것으로 사료된다.

Keywords

References

  1. Berrada, H., Borrull, F., Font, G., and Marce, R. M. : Determination of macrolide antibiotics in meat and fish using pressurized liquid extraction and liquid chromatography-mass spectrometry. J. Chromatogr. A. 1208, 83-89 (2008). https://doi.org/10.1016/j.chroma.2008.08.107
  2. McClinchey, T. A., Rafter, P. A., Regan, F., and McMahon, G. P.: A review of analytical methods for the determination of aminoglycoside and macrolide residues in food matrices. Anal. Chim. Acta. 624, 1-15 (2008). https://doi.org/10.1016/j.aca.2008.05.054
  3. Official Journal of the European Union, (EU) No 37/2010, December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin.
  4. USDA Foreign Agricultural Service. Veterinary drug MRL database. http://www.mrldatabase.com/default.cfm.
  5. Tao, Y., Yu, G., Chen, D., Pan, Y., Liu, Z., Wei, H., Peng, D., Huang, L., Wang, Y., and Yuan, Z.: Determination of 17 macrolide antibiotics and avermectins residues in meat with accelerated solvent extraction by liquid chromatography-tandem mass spectrometry. J. Chromatogr. B. Analyt. Technol. Biomed. Life. Sci. 897, 64-71 (2012). https://doi.org/10.1016/j.jchromb.2012.04.011
  6. Codex guidelines for the establishment of a regulatory programme for control of veterinary drug residues in foods (CAC/ GL 16) (1993).
  7. Validation guidelines, Codex committee on residues of veterinary drugs in foods, 11th session (1998).
  8. Le Bizec, B., Pinel, G., and Antignac, J. P.: Options for veterinary drug analysis using mass spectrometry. J. Chromatogr. A. 1216, 8016-8034 (2009). https://doi.org/10.1016/j.chroma.2009.07.007
  9. Pol-Hofstad, I., Driessen-Van Lankveld, W., Tomassen, M., De Jong, J., and Van Egmond, H. : Collaborative study of a microbiological screening method (three-plate) for the banned antimicrobial growth promotors tylosin, virginiamycin, spiramycin, zinc bacitracin and avoparcin in animal feed. Food. Addit. Contam. 25, 1465-1474 (2008). https://doi.org/10.1080/02652030802406201
  10. Vincent, U., Gizzi, G., von Holst, C., De Jong, J., and Michard, J. : Validation of an analytical method for the determination of spiramycin, virginiamycin and tylosin in feeding-stuffs by thin-layer chromatography and bio-autography. Food. Addit. Contam. 24, 351-359 (2007). https://doi.org/10.1080/02652030601075470
  11. Stobba-Wiley, C. M., Chang, J. P., Elsbury, D. T., Moran, J. W., Turner, J. M., Readnour, R. S., Stobba-Wiley, C. M., Chang JP, Elsbury DT, Moran, J. W., Turner, J. M., and Readnour, R. S.: Determination of tilimicosin residues in chicken, cattle, swine, and sheep tissues by liquid chromatography. J. AOAC. Int. 83, 837-846 (2000).
  12. Zhang, Y., Jiang, H., Jin, X., Shen, Z., Shen, J., Fu, C., and Guo, J.: Residue depletion of timicosin in chicken tissues. J. Agric. Food. Chem. 52, 2602-2605 (2004). https://doi.org/10.1021/jf035515z
  13. Maher, H. M., Youssef, R. M., Khalil, R. H., and El-Bahr, S. M.: Simultaneous multiresidue determination of metronidazole and spiramycin in fish muscle using high performance liquid chromatography with UV detection. J. Chromatogr. B. 876, 175-181 (2008). https://doi.org/10.1016/j.jchromb.2008.10.033
  14. Wang, J., Leung, D., and Butterworth, F.: Determination of five macrolide antibiotic residues in eggs using liquid chromatography/ electrospray ionization tandem mass spectrometry. .J Agric. Food. Chem. 53, 1857-1865 (2005). https://doi.org/10.1021/jf048414p
  15. Leal, C., Codony, R., Compano, R., Granados, M., and Prat, M. D.: Determination of macrolide antibiotics by liquid chromatography. J. Chromatogr. A. 910, 285-290 (2001). https://doi.org/10.1016/S0021-9673(00)01231-0
  16. Horie, M., Takegami, H., Toya, Kazuo., and Nakazawa, H.: Determination of macrolide antibiotics in meat and fish by liquid chromatography-electrospray mass spectrometry. Anal. Chim. Acta. 492, 187-197 (2003). https://doi.org/10.1016/S0003-2670(03)00891-2
  17. Abend, A. M., Chung, L., McCollum, D. G., and Wuelfing, W. P.: Development and validation of an automated extraction method (accelerated solvent extraction) and a reverse-phase HPLC analysis method for assay of ivermectin in a meatbased chewable formulation. J. Pharm. Biomed. Anal. 31, 177-183 (2003).
  18. Hwang, L. H., Yun, E. S., Kim, Y. J., Kim, D.E., Yang, Y.M., Lee, J. H., and Lee, B.D.: Study on analytical method of residual macrolide antibiotic in livestock products by LC/MS. Korean. J. Vet. Serv. 25, 221-227 (2002).
  19. Horie, M., Saito, K., Hoshino, Y., Nose, N., and Nakazawa, H.: Determination of tylosin in chicken, pork and beef by highperformance liquid chromatography. EISEI KAGAKU. 34, 128-134 (1988). https://doi.org/10.1248/jhs1956.34.128
  20. Hwang, Y. H., Lim, J. H., Park, B. K., and Yun, H. I.: Simultaneous determination of various macrolides by liquid chromatography/ mass spectrometry. J Vet.Sci. 3, 103-108 (2002).

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