Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Simultaneous Determination of Quinolones in Flatfish and Egg Using liquid Chromatography with Fluorescence Detection

액체크로마토그래피를 이용한 광어 및 계란 중 퀴놀론계의 동시분석법 개발

  • Lee, Sang-Hee (Food Analysis Center, Korea Food Research Institute) ;
  • Shim, You-Shin (Food Analysis Center, Korea Food Research Institute) ;
  • Kim, Hyun-Ju (Food Analysis Center, Korea Food Research Institute) ;
  • Choi, Yoon-Hee (Food Analysis Center, Korea Food Research Institute) ;
  • Shin, Dong-Bin (Food Analysis Center, Korea Food Research Institute)
  • 이상희 (한국식품연구원, 식품분석센터) ;
  • 심유신 (한국식품연구원, 식품분석센터) ;
  • 김현주 (한국식품연구원, 식품분석센터) ;
  • 최윤희 (한국식품연구원, 식품분석센터) ;
  • 신동빈 (한국식품연구원, 식품분석센터)
  • Published : 2008.12.31
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Abstract

An analytical method for the simultaneous determination of nine quinolones (QNs) namely, marbofloxacin, norfloxacin(IS), ciprofloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine in flatfish and egg was developed and validated using liquid chromatography with fluorescence detection (LC-FD). The samples were extracted using a traditional liquid-liquid extraction process; deproteinization was accomplished by the addition of trichloroacetic acid and acetonitrile (ACN), and defatting was performed with hexane. Chromatographic separation was achieved on a reverse phase C8 column with gradient elution using a mobile phase of 200 mM ammonium acetate buffer (pH 4.5) and ACN. The proposed method was validated according to the CODEX guideline. Mean recoveries of QNs from flatfish and egg were 89.6-106.5% with relative standard deviations (RSDs) below 15% at three different concentrations of 50, 100 and $500{\mu}g/kg$. Linearity was obtained with a correlation coefficient ($r^2$) of 0.9989-1.0000. The LOD for the investigated QNs was $1-16{\mu}g/kg$ depending on flatfish and egg. The present method can be applied simultaneously to determine QNs in muscle of flatfish and egg.

본 논문에서는 식품 중 퀴놀론계(QNs) 합성항균제(marbofloxacin, norfloxacin, danofloxacin, ciprofloxacin, enrofloxacin, difloxacin, sarafloxacin, oxolinic acid 그리고 flumequine) 9종을 분석하기 위하여, 액-액 추출 과정을 거쳐서 형광검출기가 장착된 액체크로마토그래피를 이용하여 QNs를 효율적으로 동시 분석하는 방법을 확립하였다. 컬럼은 Zorbax Eclipse XDB-C8($150\;mm{\times}4.6\;mm,\;5{\mu}m$), 이동상 용매는 200mM ammonium acetate buffer (pH 4.5)와 ACN로 기울기 용리를 사용하였으며, 유속은 1.5 ml/min, 그리고, 주입량은 $10{\mu}l$로 설정하여 분석하였다. 확립된 분석조건으로, 표준검정 곡선은 $10-500{\mu}g/kg$의 농도범위에서 상관계수가 0.9989 이상의 양호한 직선성을 나타내었으며, 회수율은 50, 100 그리고 $500{\mu}g/kg$의 농도에서 89.6-106.5%로, 향상된 추출효율을 나타내었다. 검출한계는 $1-16{\mu}g/kg$이었고, 정량한계는 $3-47{\mu}g/kg$이었으며, 일내(intra-day)와 일간(inter-day) 정밀도(CV%)는 0.2-15.0%, 0.5-11.7%이었다. 따라서, 확립된 분석방법은 광어 및 계란 중의 QNs을 효과적으로 분석하는데 이용될 수 있을 것으로 사료된다.

Keywords

References

  1. Drlica, K., and Zhao, X. : DNA gyrase, topoisomerase IV, and the 4-quinolones. Microbiol. Mol. Biol. Rev. 61, 377-92 (1997)
  2. Heo, G. J., Shin, K.S., and Lee, M. H. : Diseases of aquaculture animals and prevention of drug residues. Kor. J. Food. Hygiene. 7, 7-19 (1992)
  3. Anonymous 2004. Consolidated version of the Annexes I to IV of Council Regulation no. 2377/90. Updated 22/12/2004. Available: http://pharmacos.eudra.org/F2/mrlconspdef. Accessed May 29 (2006)
  4. Codex guidelines for the establishment of a regulatory programme for control of veterinary drug residues in foods (CAC/GL 16) (1993)
  5. Validation guidelines, Codex committee on residues of veterinary drugs in foods, 11th session (1998)
  6. Okerman, L. Noppe, H. Cornet, V., and De Zutter, L. : Microbiological detection of 10 quinolone antibiotic residues and its application to artificially contaminated poultry samples. Food. Addit. Contam. 24, 252-7 (2007) https://doi.org/10.1080/02652030600988020
  7. Cho, H. J., Abd El-Aty, A. M., Goudah, A., Sung, G. M., Seo, D. C., Kim, J. S., Shim, J. H., Jeong, J. Y., Lee, S. H., and Shin, H. C. : Monitoring of fluoroquinolone residual levels in chicken eggs by microbiological assay and confirmation by liquid chromatography. Biomed. Chromatogr. 22, 92-99 (2008) https://doi.org/10.1002/bmc.900
  8. Watanabe, H., Satake, A., Kido, Y., and Tsuji, A. : Monoclonal- based enzyme-linked immunosorbent assay and immunochromatographic assay for enrofloxacin in biological matrices. Analyst. 127, 98-103 (2002) https://doi.org/10.1039/b109427k
  9. Hassouan, M. K., Ballesteros, O., Taoufiki, J., Vilchez, J. L., Cabrera-Aguilera, M., and Navalon, A. : Meltiresidue determination of quinolones in eggs of laying hens by liquid chromatography with fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci. 852, 625-630 (2007) https://doi.org/10.1016/j.jchromb.2006.12.039
  10. Zeng, Z., Dong, A., Yang, G., Chen, Z., and Huang, X. : Simultaneous determination of nine fluoroquinolones in egg white and egg yolk by liquid chromatography with fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci. 821, 202-9 (2005) https://doi.org/10.1016/j.jchromb.2005.05.007
  11. Karbiwnyk, C. M., Carr, L. E., Turnipseed, S. B., Andersen, W. C., and Miller, K. E. : Determination of quinolone residues in shrimp using liquid chromatography with fluorescence detection and residue confirmation by mass spectrometry. Anal. Chim. Acta. 596, 257-63 (2007) https://doi.org/10.1016/j.aca.2007.06.018
  12. Schneider, M. J., Darwish, A. M., and Freeman, D. W. : Simultaneous multiresidue determination of tetracyclines and fluoroquinolones in catfish muscle using high performance liquid chromatography with fluorescence detection. Anal. Chim. Acta. 586, 269-74 (2007) https://doi.org/10.1016/j.aca.2006.09.025
  13. Roudaut, B., and Yorke, J, C. : High-performance liquid chromatographic method with fluorescence detection for the screening and quantification of oxolinic acid, flumequine and sarafloxacin in fish. J Chromatogr B Analyt Technol Biomed Life Sci. 780, 481-5 (2002) https://doi.org/10.1016/S1570-0232(02)00641-4
  14. Christodoulou, E. A., Samanidou, V. F., and Papadoyannis, I. N. : Validation of an HPLC-UV method according to the European Union Decision 2002/657/EC for the simultaneous determination of 10 quinolones in chicken muscle and egg yolk. J Chromatogr B Analyt Technol Biomed Life Sci. 859, 246-55 (2007) https://doi.org/10.1016/j.jchromb.2007.10.009
  15. Samanidou, V., Evaggelopoulou, E., Trotzmuller, M., Guo, X., and Lankmayr, E. : Multi-residue determination of seven quinolones antibiotics in gilthead seabream using liquid chromatography-tandem mass spectrometry. J Chromatogr A. 1203, 115-23 (2008) https://doi.org/10.1016/j.chroma.2008.07.003
  16. Barbosa, J., Berges, R., Sanz-Nebot, V. : Retention behaviour of quinolone derivatives in high-performance liquid chromatography effect of p H and evaluation of ionization constant. J. Chromatogr. A. 823, 411-422 (1998) https://doi.org/10.1016/S0021-9673(98)00428-2
  17. Gigosos, P. G., Revesado, P. R., Cadahía, O., Fente, C. A., Vazquez, B. I., Franco, C. M., and Cepeda, A. : Determination of quinolones in animal tissues and eggs by high-performance liquid chromatography with photodiode-array detection. J Chromatogr A. 871, 31-6 (2000) https://doi.org/10.1016/S0021-9673(99)01048-1