Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
권호 표지
DOI QR코드

DOI QR Code

Simultaneous Determination of Penicillin Antibiotics in Meat using Liquid Chromatography Tandem Mass Spectrometry

LC-MS/MS를 이용한 육류 중 페니실린계 항생제 8종의 동시분석 및 적용성 검증

  • Kim, Myeong-Ae (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Yoon, Su-Jin (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Kim, MeeKyung (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Cho, Yoon-Jae (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Choi, Sun-Ju (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Chang, Moon-Ik (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Lee, Sang-Mok (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Kim, Hee-Jeong (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Jeong, Jiyoon (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Rhee, Gyu-Seek (Pesticide and Veterinary Drug Residues Division, National Institute of Food & Drug Safety Evaluation) ;
  • Lee, Sang-Jae (Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation)
  • 김명애 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 윤수진 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 김미경 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 조윤제 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 최선주 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 장문익 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 이상목 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 김희정 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 정지윤 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 이규식 (식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 이상재 (식품의약품안전평가원 식품위해평가부)
  • Received : 2014.02.12
  • Accepted : 2014.05.16
  • Published : 2014.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

The objective of this study was to develop a simultaneous method of 8 penicillin antibiotics including amoxicillin, ampicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin, penicillin G and penicillin V in meat using LC-MS/MS. The procedure involves solid phase extraction with HLB cartridge and subsequent analysis by LC-MS/MS. To optimize MS analytical condition of 8 compounds, each parameter was established by multiple reaction monitoring in positive ion mode. The chromatographic separation was achieved on a $C_{18}$ column with a mobile phase of 0.05% formic acid and 0.05% formic acid in acetonitrile at a flow rate of 0.2 mL/min for 20 min with a gradient elution. The developed method was validated for specificity, linearity, accuracy and precision in beef, pork and chicken. The recoveries were 71.0~106%, and relative standard deviations (RSD) were 4.0~11.2%. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.003~0.008 mg/kg and 0.01~0.03 mg/kg, respectively, that are below maximum residue limit (MRL) of the penicillins. This study also performed survey of residual penicillin antibiotics for 193 samples of beef, pork and chicken collected from 9 cities in Korea. Penicillins were not found in all the samples except a sample of pork which contained cloxacillin (concentration of 0.08 mg/kg) below the MRL (0.3 mg/kg).

본 연구에서는 LC-MS/MS를 이용하여 페니실린계 동물용의약품 8종, 암피실린, 아목시실린, 클록사실린, 디클록사실린, 나프실린, 옥사실린, 페니실린 G, 페니실린 V의 동시분석법을 확립하고 잔류실태조사를 수행하였다. 피페라실린을 내부표준물질으로 적용하였으며, 이동상으로는 0.05% 포름산 용액와 0.05% 포름산 아세토니트릴 용액을 gradient mode로 사용하였고 $C_{18}$ 칼럼을 이용하여 분석하였다. 각 분석물질은 MRL의 0.25, 0.5, 1, 2, 4, 8배 농도범위에서 검량선을 작성하여 $R^2{\geq}0.99$의 직선성 및 재현성을 확인하였다. 소고기, 돼지고기, 닭고기에 대하여 MRL의 0.5, 1, 2배인 세 가지 농도에서 6회 반복으로 분석을 실시한 결과, 평균 회수율은 71.0~106%, 상대표준편차는 4.0~11.2%, LOD는 0.003~0.008 mg/kg, LOQ는 0.01~0.03 mg/kg이었다. 국내 유통 소고기, 돼지고기, 닭고기에서 페니실린계 항생제 8종의 분석결과, 총 193건 중 경기도 지역의 국내산 소고기 1건에 대해서 클록사실린이 MRL 이하인 0.08 mg/kg이 검출되었으며, 나머지 192건의 검체에서는 불검출로 분석되었다. 본 연구를 통하여 확립된 페니실린계 동물용의약품 8종 동시분석법은 식육에 대한 페니실린계 동물용의약품의 안전관리 체계 방안을 제시하기 위한 기초자료로 활용 가능한 것으로 사료된다.

Keywords

References

  1. Shin H.C., Park K.H.: Veterinary drugs residue monitoring. KFDA, 9-61 (2006).
  2. Gil G.H., Ko K.Y., Lee Y.J., Park H.J., Lee K.S.: Simultaneous determination of tetracyclines and sulfonamides residues in pork and flatfish using Matrix Solid Phase Dispersion (MSPD) extraction and HPLC. Korean J. Environ. Agr., 28, 266-273 (2009). https://doi.org/10.5338/KJEA.2009.28.3.266
  3. Shin H.C., Han S.B., Park K.H.: Residue monitoring and analytical method development of veterinary drugs permitted and not-permitted to use in food producing animals and fishes. KFDA, 11-13 (2007).
  4. Lee M.H., Yun S.M., Lee K.J., Kim H.R., Son S.W.: Regulation and issue of veterinary drug ; Overview of the herbals for animal drug use and quality specifications. Korean J. Vet. Public Health, 30, 147-151 (2006).
  5. Turnipseed S.B., Roybal J.E., Andersen W.C., Kuck L.R.: Analysis of avermectin an moxidectin residues in milk by liquid chromatography-tandem mass spectrometry using an atmospheric pressure chemical ionization/atmospheric pressure photoionization source. Anal. Chim. Acta., 529, 159-165 (2005). https://doi.org/10.1016/j.aca.2004.07.061
  6. Kim H.Y., Chung S.Y., Choi S.H., Lee J.S., Choi I.S., Cho M.J., Shin M.S., Song J.S, Choi J.C., Park H.O., Ha S.C., Shin I.S., Seo E.C.: Monitoring of veterinary drug residues in foods produced in Korea. Korean J. Food Sci. Technol., 42, 653-663 (2010).
  7. http://www.qia.go.kr/viewwebQiaCom.do?id=34070&type=6_18_1bdsm: 축산 항생제 사용량 2년 연속 최저. 농림축산검역본부 (2013).
  8. Hong J.H.: Method development and monitoring on residues of veterinary drugs in foods. KFDA, 11-35 (2012).
  9. Kim M.K., Cho B.H., C.M., Kim D.G., Yune S.Y., Shin J.Y., Bong Y.H., Kang J.W., Kim M.A., Son S.W.: Chemical Residues and Contaminants in Foods of Animal Origin in Korea during the Past Decade. J. Agr Food Chem., 61, 2293-2298 (2013). https://doi.org/10.1021/jf3046297
  10. Anfossi1 P., Zaghini1 A., Grassigli G., Menotta S., Fedrizzi G.: Relative oral bioavailability of microgranulated amoxicillin in pigs. J. Vet. Pharmacol. Ther., 25, 329-334 (2002). https://doi.org/10.1046/j.1365-2885.2002.00428.x
  11. Song J.Y, Hu S.J., Joo H.J., Kim M.O., Hwang J.B., Han Y.J., Kwon Y.J., Kang S.J., Cho D.H.: Residue analysis of penicillines in livestock and marine products. Anal. Sci. Tech., 25, 257-264 (2012). https://doi.org/10.5806/AST.2012.25.4.257
  12. Macarov C.A., Tong L., Martinez-Huelamo M., Hermo M.P., Chirila E., Wang Y.X., Barron D., Barbosa J.: Multi residue determination of the penicillins regulated by the European Union, in bovine, porcine and chicken muscle, by LC-MS/ MS. Food Chem., 135, 2612-2621 (2012). https://doi.org/10.1016/j.foodchem.2012.06.126
  13. Boison JO, Keng LJ, MacNeil JD.: Analysis of penicillin G in milk by liquid chromatography. J. AOAC Int., 77, 565-570 (1994).
  14. Tyczkowska K.L., Voyksner R.D., Straub R.F., Aronson A.L.: Simultaneous multiresidue analysis of beta-lactam antibiotics in bovine milk by liquid chromatography with ultraviolet detection and confirmation by electrospray mass spectrometry. J. AOAC Int., 77, 1122-1131 (1994).
  15. Ang C.Y.W., Luo W.: Rapid determination of ampicillin in bovine milk by liquid chromatography with fluorescence detection. J. AOAC Int., 80, 25-30 (1997).
  16. Abjean J.P., Lahogue V.: Planar chromatography for quantitative determination of ampicillin residues in milk and muscle. J. AOAC Int., 80, 1171-1176 (1997).
  17. Yoon S.B., Im C.U.: $\beta$-lactamase inhibitory activity and comparative activity of 6-benzothiazole penicillin derivatives in combination with $\beta$-lactam antibiotics. Pharm. Soc. Korea, 52, 306-310 (2008).
  18. Cunha B.A.: Antibiotic side effects. Med. Clin. N. Am., 85, 149-185 (2001). https://doi.org/10.1016/S0025-7125(05)70309-6
  19. 서울대학교약학교육연수원: 의약품집. 서울대학교출판부, 258 (2002).
  20. 의학교육연수원: 가정의학. 서울대학교출판부, 서울, 1129 (1998).
  21. Rhee G.S.: Method improvement and monitoring on residues of veterinary drugs in foods Ministry of Food and Drug Safety, 13 (2013).
  22. Choi D.M.: 2009 Veterinary drug residues monitoring. KFDA, 4-14 (2009).
  23. Kukusamudea C., Burakhama R., Chailapakulb O., Srijaranai S.: High performance liquid chromatography for the simultaneous analysis of penicillin residues in beef and milk using ion-paired extraction and binary water-acetonitrile mixture. Talanta, 92, 38-44 (2012). https://doi.org/10.1016/j.talanta.2012.01.020
  24. Ito Y., Ikai Y., Oka H., Kagami T., Takeba K.: Application of ion-exchange cartridge clean-up in food analysis: II. Determination of benzylpenicillin, phenoxymethylpenicillin, oxacillin, cloxacillin, nafcillin and dicloxacillin in meat using liquid chromatography with ultraviolet detection. J.Chromatogr. A, 855, 247-253 (1999). https://doi.org/10.1016/S0021-9673(99)00695-0
  25. Ito Y., Ikai Y., Oka H., Matsumoto H., Kagami T., Takeba K.: Application of ion-exchange cartridge clean-up in food analysis: III. Determination of benzylpenicillin, phenoxymethylpenicillin, oxacillin, cloxacillin, nafcillin and dicloxacillin in bovine liver and kidney by liquid chromatography with ultraviolet detection. J. Chromatogr. A, 880, 85-91 (2000). https://doi.org/10.1016/S0021-9673(00)00201-6
  26. Goto T., Ito Y., Yamada S., Matsumoto H., Oka H.: Highthroughput analysis of tetracycline and penicillin antibiotics in animal tissues using electrospray tandem mass spectrometry with selected reaction monitoring transition. J. Chromatogr. A, 1100, 193-199 (2005). https://doi.org/10.1016/j.chroma.2005.09.056
  27. Chico J., Rubies A., Centrich F., Companyo R., Prat M.D., Granados M.: High-throughput multiclass method for antibiotic residue analysis by liquid chromatography-tandem mass spectrometry. J. Chromatogr. A, 1213, 189-199 (2008). https://doi.org/10.1016/j.chroma.2008.10.023
  28. Ito Y., Goto T., Oka H., Matsumoto H., Takeba K.: Application of ion-exchange cartridge clean-up in food analysis: VI. Determination of six penicillins in bovine tissues by liquid chromatography-electrospray ionization tandem mass spectrometry. J. Chromatogr. A, 1042, 107-111 (2004). https://doi.org/10.1016/j.chroma.2004.05.057
  29. Junza A., Amatya R., Barron D., Barbosa J.: Comparative study of the LC-MS/MS and UPLC-MS/MS for the multiresidue analysis of quinolones, penicillins and cephalosporins in cow milk, and validation according to the regulation 2002/657/EC. J. Chromatogr. B, 879, 2601-2610 (2011). https://doi.org/10.1016/j.jchromb.2011.07.018
  30. Hsieh S.H., Huang H.Y., Lee S.: Determination of eight penicillin antibiotics in pharmaceuticals, milk and porcine tissues by nano-liquid chromatography. J. Chromatogr. A, 1216, 7186-7194 (2009). https://doi.org/10.1016/j.chroma.2009.05.080
  31. Hwang L.H., Yun E.S., Kim H.J., Kim Y.J., Jung H.K., Han I.K., Lee B.D.: A study on simultaneous determination of residual penicillin G and chloramphenicol in livestock products by high performance liquid chromatography. Korean J. Vet. Serv., 23, 1-8 (2000).
  32. Lim C.M., Kwon H.J., Cho B.H., Park S.J., Chung G.S., Son S.W.: Confirmatory assay for the simultaneous detection of eight penicillin antibiotics in swine muscle and kidney by Liquid chromatography coupled with tandem mass spectrometry. Korean J. Vet. Public Health, 35, 275-283 (2011).
  33. 축수산물 유해물질 분석법 편람. 식품의약품안전평가원(2013).
  34. Codex Committee on Pesticide Residues: Guidelines on Good Laboratory Practice in Pesticide Residue Analysis, CODEX, CAC/GL 40-1993 (2010).