Journal of fish pathology (한국어병학회지)

The Korean Society of Fish Pathology (한국어병학회)
권호 표지
DOI QR코드

DOI QR Code

Analytical method study for cephalexin with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) applicable for residue studies in the whiteleg shrimp Litopenaeus vannamei

흰다리새우(Litopenaeus vannamei)에서 cephalexin의 잔류농도 연구를 위한 LC-MS/MS 분석법 연구

  • Yang, Chan Yeong (Departments of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University) ;
  • Bae, Jun Sung (Departments of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University) ;
  • Lee, Chae Won (Departments of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University) ;
  • Jeong, Eun Ha (Departments of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University) ;
  • Lee, Ji-Hoon (National Fishery Products Quality Management Service) ;
  • Bak, Su-Jin (National Fishery Products Quality Management Service) ;
  • Choi, Sang-Hoon (Departments of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University) ;
  • Park, Kwan Ha (Departments of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University)
  • 양찬영 (군산대학교 해양과학대학 수산생명의학과) ;
  • 배준성 (군산대학교 해양과학대학 수산생명의학과) ;
  • 이채원 (군산대학교 해양과학대학 수산생명의학과) ;
  • 정은하 (군산대학교 해양과학대학 수산생명의학과) ;
  • 이지훈 (국립수산물품질관리원) ;
  • 박수진 (국립수산물품질관리원) ;
  • 최상훈 (군산대학교 해양과학대학 수산생명의학과) ;
  • 박관하 (군산대학교 해양과학대학 수산생명의학과)
  • Received : 2021.05.03
  • Accepted : 2021.06.08
  • Published : 2021.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Cephalexin, a semi-synthetic cephalosporin antibiotic, has long been used in fish aquaculture in various countries under legal authorization. The drug is thus widely available for use in other aquatic species except fishes like the crustacean whiteleg shrimp. This study aims to develop a sensitive method for laboratory residue studies to adopt in withdrawal period determinations. Through repeated trials from the existing methods developed for other food animal tissues, it was possible to achieve a sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. The results showed that at a concentration of 0.1 mg/kg, the recovery rate was 81.79%, and C.V. value was 8.2%, which meet the recovery rate and C.V. recommended by Codex guideline. After satisfactory validation of analytical procedures, applicability to the shrimp tissue was confirmed in experimentally cephalexin-treated whiteleg shrimp. As a result, most muscle samples were detected below the limit of quantification (0.05 mg/kg) after day 3, and most hepatopancreas samples were detected below the limit of quantification after day 14. In particular, the limit of quantification 0.05 ppm with the presently developed method suggests sufficient sensitive over the current legal maximum residue limit of 0.2 mg/kg set for fishes.

세팔로스포린계열의 반합성 항생제인 세팔렉신은 법적 허가를 받은 여러 나라의 양식장에서 사용되어 왔다. 따라서 이 항생제는 갑각류 중 흰다리새우와 같은 어류를 제외한 다른 수생 생물 종에서 광범위하게 사용할 수 있다. 본 연구는 흰다리새우에서 세팔렉신의 잔류기간 설정을 위해 잔류분석법을 개발하였고, 이를 바탕으로 잔류실험을 수행하였다. 분석법 개발을 위하여 이미 존재하고 있는 여러 식용동물 조직에서의 분석법을 바탕으로 반복 시도하여 고성능 액체 크로마토그래피와 질량분석기(high-performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)를 이용한 분석법을 개발하였다. 그 결과, 0.1 mg/kg의 농도에서 회수율이 81.79%, C.V.값은 8.2%로 Codex guideline에서 권장하는 회수율 및 C.V.에 충족하는 결과를 나타내었다. 개발한 분석법의 적정성을 검증 후, 흰다리새우에 세팔렉신을 경구투여 한 뒤 잔류량을 분석하여 흰다리새우 조직에서의 적용 가능성을 확인하였다. 그 결과, 투여 종료 3일 후에 대부분의 근육 시료에서 정량한계(0.05 mg/kg)이하로 검출되었고, 투여 종료 14일 후에 대부분의 간췌장 시료에서 정량한계 이하로 검출되었다. 특히 본 연구에서 개발한 분석법에서 설정된 정량한계(0.05 ppm)는 어류에서 법적으로 설정된 잔류허용 기준 0.2 mg/kg에 대해 충분한 감도를 나타낸다.

Keywords

References

  1. Benarab, N. and Fangninou, F.F.: The issues of antibiotics: cephalexin antibiotic as emerging environmental contaminant. J. Sci. Res. Pub., 10:306-318, 2020.
  2. Chae, W.S., Lee, S.J., Son, S.E., Kim, S. and Lee, H.J.: Analytical method for determination of cephalexin in bovine edible tissues using liquid chromatography coupled to tandem mass spectrometry. J. Food Hyg. Saf., 33:58-64, 2018. https://doi.org/10.13103/JFHS.2018.33.1.58
  3. Codex Alimentarius Commission: Codex Guidelines for the Establishment of a Regulatory Programme for Control of Veterinary Drug Residues in Foods (CAC/GL-16). http://www.fao.org/fao-who-codex-alimentarius/home/pt/, 1993.
  4. Faroongsarng, D., Chiayvareesajja, S., Chandumpai, A., Theapparat, Y.: Hepatopancreatic and muscular distribution of oxytetracycline antibiotics in farmed pacific white shrimp (Penaeus vannamei): a physiological-based pharmacokinetic model approach. Aquacult. Res., 41:143-152, 2009. https://doi.org/10.1111/j.1365-2109.2009.02324.x
  5. Jung, H.N., Park, D.H., Yoo, K.H., Cho, H.J., Shim, J.H., Shin, H.C. and Abd El-Aty A.M.: Simultaneous quantification of 12 veterinary drug residues in fishery products using liquid chromatography-tandem mass spectrometry. J. Food Chem., 348:1-10, 2021.
  6. KFDA (Korea Food & Drug Administration): Pesticides and veterinary drugs information. https://www.foodsafetykorea.go.kr/residue/main.do, 2021a
  7. KFDA (Korea Food & Drug Administration): Food code. http://www.foodsafetykorea.go.kr/foodcode/01_02.jsp?idx=263, 2021b
  8. Kim, M.R., Choi, Y.H., Kim, D.H., Kim, Y.S. and Lee, J.H.: Monitoring for cephalosporin residues in raw meat in Seoul. J. Vet. Serv., 38:259-264, 2015. https://doi.org/10.7853/kjvs.2015.38.4.259
  9. Kim, J.W., Cho, M.Y., Jee, B.Y., Park, M.A and Kim, N.Y.: Administration and use of aquaculture drugs in Korea. J. Fish Pathol., 27:67-75, 2014. https://doi.org/10.7847/jfp.2014.27.1.067
  10. Kim, N.E. and Kim, D.H.: Acute hepatopancreatic necrosis disease of shrimp and import health measures. J. Fish Pathol., 28:1-7, 2015. https://doi.org/10.7847/jfp.2015.28.1.001
  11. Kim, J.W., Lee, H.N., Jee, B.Y., Woo, S.H., Kim, Y.J. and Lee, M.K.: Monitoring of the mortalities in the aquaculture farms of South Korea. J. Fish Pathol., 25:271-277, 2012. https://doi.org/10.7847/jfp.2012.25.3.271
  12. Kim, C.S., Ryu, H.D., Chung, E.G., Kim, Y.S. and Rhew, D.H.: Determination of antibiotic residues:I. Extraction and clean-up Methods for solid samples: a review. J. Kor. Soc. Water Environ., 32:600-627, 2016. https://doi.org/10.15681/KSWE.2016.32.6.600
  13. Kim, Y.J., Seo, J.S., Park, J.O., Jeong, A.R. and Lee, J.H.: Monitoring of aquatic medicine managements in South Korea. J. Fish Pathol., 32:37-43, 2019. https://doi.org/10.7847/JFP.2019.32.1.037
  14. Kim, S.K., Shim, N.Y., Jang, J.W., Jun, J.C., Kim, S.K. and Shin, Y.K.: Effect of acclimation methods on physiological status of white shrimp, Litopenaeus vannamei larvae to low salinities. J. Kor. Soc. Environ. Biol., 35:6-12, 2017. https://doi.org/10.11626/KJEB.2017.35.1.006
  15. KOSIS (Korean Statistical Information Service): Statistics for each variety of fishery. https://kosis.kr/statisticsList/statisticsListIndex.do?vwcd, 2021.
  16. Lee, C.W.: An analytical method development of cephalexin in whiteleg shrimp (Litopenaeus vannamei) using LC-MS/MS and its application in residue concentration studies. Thesis for Master's Degree, Kunsan National University, Korea, February 2020.
  17. Lee, J.H.: Development of cefadroxil analysis and its application to residuce analysis in fishes by LC-MS/MS, Thesis for Master's Degree, Kunsan National University, Korea, February 2016.
  18. Lee, K.B., Kim, N.W., Kim, D.U., Sin, M.H. and Lee, M.Y.: Monitoring of veterinary drug residues in cultured fishery products in Chungcheongnam-do Province. J. Food Hyg. Saf., 35:447-451, 2020. https://doi.org/10.13103/JFHS.2020.35.5.447
  19. Lee, Y.M. and Zoh, K.D.: Recent analysis of veterinary drug residues in food. J. Pub. Health, 53:27-37, 2016.
  20. Lim, J.H., Jang, B.S., Park, B.K. and Yun, H.I.: Residue depletion of cephalexin in the flounder (Paralichthys olivaceus). J. Vet. Res., 43:435-438, 2003.
  21. Lim, S.K.: Careful use of cephalosporin antibiotics. J. Kor. Vet. Med. Assoc., 50:734-738, 2014.
  22. Ma, R., Huang, L., Wei, W., Wang, Y., Zou, X., Zhou, J., Li, X. and Fang, W.: Pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin in Pacific white shrimp Litopenaeus vannamei after multiple-dose oral administration. Fish. Sci., 84:869-876, 2018. https://doi.org/10.1007/s12562-018-1229-y
  23. NIFDS (National Institute of Food and Drug Safety): A casebook of excellence in MFDS research and development projects in 2016. https://www.nifds.go.kr/brd/m_347/view.do?seq=10503, 2016.
  24. Song, J.Y., Hu, S.J., Joo, H.J., Kim, M.O., Hwang, J.B., Han, Y.J., Kwon, Y.J., Kang, S.J. and Cho, D.H.: Residue analysis of penicillines in livestock and marine products. J. Anal. Sci. Technol., 25:257-264, 2012. https://doi.org/10.5806/AST.2012.25.4.257
  25. Takamuku, T., Yamaguchi, A., Matsue, D,. Tabata, M., Kumamoto, M., Nishimoto, J., Yoshida, K., Yamaguchi. T., Nagao, M., Otomo, T. and Adachi, T.: Large-angle X-ray scattering and small-angle neutron scattering study on phase separation of acetonitrile-water mixtures by addition of NaCl. J. Physi. Chem. B, 105:6236-6245, 2001. https://doi.org/10.1021/jp003011n
  26. unlusayin, M., Erdilal, R., Gumus, B. and Gulyavuz, H.: The effects of salt boiling on protein loss of Penaeus semisulcatus. J. Fish. Aquat. Sci., 10:75-79, 2010.
  27. Wang, L., Li, W., Wang, Y., Wang, C., Ye, D., Zhou, L., Hu, X., Ke, Y. and Xia, X.: Determination of cephalexin residual level using ultra-high-performance liquid chromatography-tandem mass spectrometry: residue depletion study in swine. J. Chromatogr. B, 1124:233-238, 2019. https://doi.org/10.1016/j.jchromb.2019.06.003