한국식품위생안전성학회지 (Journal of Food Hygiene and Safety)

  • 제34권1호
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  • Pages.13-21
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  • 2019
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  • 1229-1153(pISSN)
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  • 2465-9223(eISSN)
한국식품위생안전성학회 (The Korean Society of Food Hygiene and Safety)
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LC-MS/MS를 이용한 농산물 중 Streptomycin 및 Dihydrostreptomycin 동시시험법 개발

Development of Simultaneous Analytical Method for Streptomycin and Dihydrostreptomycin Detection in Agricultural Products Using LC-MS/MS

  • 이한솔 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 도정아 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 박지수 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 박신민 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 조성민 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 신혜선 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 장동은 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 최영내 (한국식품과학연구원) ;
  • 정용현 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과) ;
  • 이강봉 (식품의약품안전처 식품의약품안전평가원 식품위해평가부 잔류물질과)
  • Lee, Han Sol (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Do, Jung-Ah (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Park, Ji-Su (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Park, Shin-Min (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Cho, Sung Min (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Shin, Hye-Sun (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Jang, Dong Eun (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Choi, Young-Nae (Korea Advanced Food Research Institute) ;
  • Jung, Yong-hyun (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Kangbong (Pesticide and Veterinary Drug Residues Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety)
  • 투고 : 2018.11.07
  • 심사 : 2018.12.04
  • 발행 : 2019.02.28
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초록

본 연구는 2018년 국내식품 중 신규 기준설정 예정 농약인 스트렙토마이신의 잔류 농약 검사를 위한 공정시험법을 마련하기 위하여 수행하였다. 스트렙토마이신은 아미노글리코사이드계 항생제로 소, 돼지, 닭 등 가축 및 가금에 동물용의약품으로 허가되어 세균성 질병치료에 사용되고 있다. 또한 우리나라에서는 감귤, 매실, 참다래, 복숭아 등에 발병하는 세균성 병해를 방지하기 위하여 농약으로 등록되어 있다. 국내에서는 스트렙토마이신의 잔류물의 정의를 스트렙토마이신과 디히드로스트렙토마이신의 합으로 정하여 관리하고 있으며 잔류허용기준은 동물용의 약품으로 사용 시 불검출부터 1.0 mg/kg까지 설정되어 있다. 농약으로 사용 시 농산물 중 잔류허용기준은 설정되어 있지 않으며 식품공전의 시험법 역시 동물용의약품에 적용되는 시험법으로 설정되어 있다. 이에 농산물 중 잔류허용기준 설정을 위한 시험법 마련이 시급하여 본 연구에서는 식품공전에서 정하고 있는 대표농산물 5종(현미, 감자, 대두, 감귤, 고추)을 포함한 잔류시험법을 개발하고자 하였다. 따라서 수용성 유기용매인 메탄올의 적용과 포름산을 이용한 pH 조절을 통한 추출법 및 HLB 카트리지를 이용한 정제법을 최적화하여 LC-MS/MS에 의한 분석법을 확립하였다. 스트렙토마이신과 디히드로스트렙토마이신의 시험법 정량한계는 0.01 mg/kg으로 PLS 제도에 맞는 잔류허용기준 수준을 만족하였다. 스트렙토마이신과 디히드로스트렙토마이신의 평균 회수율은 각각 72.0~116.5%와 72.1~116.0%이었고, 상대표준편차는 각각 12.3%와 12.5% 이하로 조사되어 이러한 결과는 국제식품규격위원회 가이드라인(CAC/GL 40-1993, 2003)의 잔류농약 분석기준 및 식품의약품안전평가원의 '식품등 시험법 마련 표준절차에 관한 가이드라인(2016)'에 적합한 수준임을 검증하였다. 따라서 본 연구에서 개발한 시험법은 농산물 중 스트렙토마이신의 잔류검사를 위한 공정시험법으로 활용할 수 있으며 유사 농산물에 대한 적용도 가능하리라 판단되어 향후 국내 농산물 중 농약의 잔류허용기준 신설 및 잔류농약 검사의 기초자료로 활용 가능할 것이다.

A method was developed for the simultaneous detection of an antibiotic fungicide, streptomycin, and its metabolite (dihydrostreptomycin) in agricultural products using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted using methanol adjusted to pH 3 using formic acid, and purified with a HLB (Hydrophilic lipophilic balance) cartridge. The matrix-matched calibration curves were constructed using seven concentration levels, from 0.001 to 0.1 mg/kg, and linearity of five agricultural products (hulled rice, potato, soybean, mandarin, green pepper), with coefficients of determination $(R^2){\geq}0.9906$, for streptomycin and dihydrostreptomycin. The mean recoveries at three fortification levels (LOQ, $LOQ{\times}10$, $LOQ{\times}50$, n = 5) were from 72.0~116.5% and from 72.1~116.0%, and relative standard deviations were less than 12.3% and 12.5%, respectively. The limits of quantification (LOQ) were 0.01 mg/kg, which are satisfactory for quantification levels corresponding with the Positive List System. All optimized results satisfied the criteria ranges requested in the Codex guidelines and the Food Safety Evaluation Department guidelines. The present study could serve as a reference for the establishment of maximum residue limits and be used as basic data for detection of streptomycin and dihydrostreptomycin in food.

키워드

참고문헌

  1. Korea Crop Protection Association (KCPA), Using guideline of crop protection agents. Available at http://www.koreacpa.org/korea/bbs/board.php?bo_table=3_3 (2018).
  2. Korean Society for Molecular and Cellular Biology, Streptomycin. Available at https://terms.naver.com/entry.nhn?docId=5569295&cid=61233&categoryId=61233 (2018).
  3. Encyclopedia of Life Science, Dihydrostreptomycin. Available at https://terms.naver.com/entry.nhn?docId=425088&cid=60261&categoryId=60261 (2014).
  4. Chemicalize, Streptomycin. Available at https://chemicalize.com/#/calculation (2018).
  5. Chemicalize, Dihydrostreptomycin. Available at https://chemicalize.com/#/calculation (2018).
  6. Ministry of Food and Drug Safety (MFDS), MRLs in Veterinary drugs. Available at http://www.foodsafetykorea.go.kr/residue/vd/mrls/list.do?currentPageNo=1&searchCode=&menuKey=2&subMenuKey=83&subChildMenuKey=&searchConsonantFlag=&searchConsonantFlag2=&searchValue2=&searchFlag=vd&searchClassLCode=&searchClassMCode=&searchClassScode=&searchValue=%EC%8A%A4%ED%8A%B8%EB%A0%99%ED%86%A0%EB%A7%88%EC%9D%B4%EC%8B%A0 (2018).
  7. Ministry of Food and Drug Safety (MFDS), MRLs in Pesticide. Available at http://www.foodsafetykorea.go.kr/residue/prd/mrls/list.do?currentPageNo=1&searchCode=&searchFoodCode=&menuKey=1&subMenuKey=161&subChildMenuKey=&searchConsonantFlag=&searchConsonantFlag2=&searchValue2=&searchFlag=prd&searchClassLCode=&searchClassMCode=&searchClassScode=&searchValue=%EC%8A%A4%ED%8A%B8%EB%A0%99%ED%86%A0 (2018).
  8. The Joint FAO/WHO Expert Committee on Food Additives (JECFA), Online Edition: "Residues of some veterinary drugs in foods and animals". Available at http://www.fao.org/food/food-safety-quality/scientific-advice/jecfa/jecfavetdrugs/details/en/c/41/ (2018).
  9. The Japan Food Chemical Research Foundation, Maximum Residue Limits (MRLs) List of Agricultural Chemicals in Foods. Available at https://db.ffcr.or.jp/front/pesticide_detail?id=29800 (2018).
  10. European Commission, EU Pesticides database. Available at http://ec.europa.eu/food/plant/pesticides/eu-pesticides-database/public/?event=pesticide.residue.CurrentMRL&language=EN (2018).
  11. United States Environmental Protection Agency (US EPA), Regulation of Pesticide Residues on Food. Available at https://www.ecfr.gov/cgi-bin/text-idx?SID=3dcd650aebb04d701750379c98587a8a&mc=true&node=se40.26.180_1245&rgn=div8 (2018).
  12. Ministry of Food and Drug Safety (MFDS), Korea food code. pp. 1449-1452 (2018).
  13. Shim Y. E., Myung S. W.: Analysis of streptomycin in honey by LC-MS/MS. Anal. Sci. Technol., 21, 424-431 (2008).
  14. Do J. A., Lee M. Y., Cho Y. J., Chang M. I., Hong J. H., Oh J. H.: Determination of streptomycin in kiwifruit samples using LC-ESI-MS/MS. Anal. Sci. Technol., 28, 299-307 (2015). https://doi.org/10.5806/AST.2015.28.4.299
  15. Ministry of Food and Drug Safety (MFDS), Korea food code. pp. 1377-1382 (2018).
  16. Bruijnsvoort, M. van, Ottink, S. J. M., Jonker, K. M., Boer, E. de: Determination of streptomycin and dihydrostreptomycin in milk and honey by liquid chromatography with tandem mass spectrometry. J. Chromatogr. A., 1058, 137-142 (2004). https://doi.org/10.1016/j.chroma.2004.07.101
  17. Bohm, D. A., Stachel, C. S., Gowik, P.: Confirmatory method for the determination of streptomycin and dihydrostreptomycin in honey by LC-MS/MS. Food Addit. Contam., 29, 189-196 (2012). https://doi.org/10.1080/19440049.2011.635347
  18. Oliveira, R. C. de, Paschoal, J. A. R., Sismotto, M., Airoldi, F. P. da S., Reyes, F. G. R.: Development and validation of an LC-APCI-MS-MS analytical method for the determination of streptomycin and dihydrostreptomycin residues in milk. J. Chromatogr. Sci., 47, 756-761 (2009). https://doi.org/10.1093/chromsci/47.9.756
  19. Gremilogianni, A. M., Megoulas, N. C., Koupparis, M. A.: Hydrophilic interaction vs ion pair liquid chromatography for the determination of streptomycin and dihydrostreptomycin residues in milk based on mass spectrometric detection. J. Chromatogr. A., 1217, 6646-6651 (2010). https://doi.org/10.1016/j.chroma.2010.04.059
  20. Kang Y. W., Joo H. J., Kim Y. S., Cho Y. J., Kim H. Y., Lee G. H., Kim M. H. Analysis and monitoring of residues of aminoglycoside antibiotics in livestock products. Korean J. Food Sci. Technol., 43, 1-5 (2011). https://doi.org/10.9721/KJFST.2011.43.1.001
  21. EU Reference Laboratory for pesticides requiring Single Residue Methods (EURL-SRM), Quick Method for the Analysis of numerous Highly Polar Pesticides in Foods of Plant Origin via LC-MS/MS involving Simultaneous Extraction with Methanol (QuPPe-Method). Version 9.3. Available at http://www.eurl-pesticides.eu/userfiles/file/EurlSRM/meth_QuPPe-PO_EurlSRM.pdf (2017).
  22. Ministry of Food and Drug Safety (MFDS), Pesticide analytical residues manual in food code. pp. 13-14 (2013).
  23. Ko A. Y., Abd El-Aty, A. M., Rahman, M. M., Jang J., Kim S. W., Choi J. H., Shim, J. H.: A modified QuEChERS method for simultaneous determination of flonicamid and its metabolites in paprika using tandem mass spectrometry. Food Chem., 157, 413-420 (2014). https://doi.org/10.1016/j.foodchem.2014.02.038
  24. Millership, J. S., Hare, L. G., Farry, M., Collier, P. S., McElnay, J. C., Shields, M. D., Carson, D. J.: The use of hydrophilic lipophilic balanced (HLB) copolymer SPE cartridges for the extraction of diclofenac from small volume paediatric plasma samples. J. Pharm. Biomed. Anal., 25, 871-879 (2001). https://doi.org/10.1016/S0731-7085(01)00367-3
  25. Jadhav, M. R., Utture, S. C., Banerjee, K., Oulkar, D. P., Sabale, R., Ahammed, S. T. P.: Validation of a residue analysis method for streptomycin and tetracycline and their food safety evaluation in pomegranate (Punica granatum L.). J. Agric. Food Chem., 61, 8491-8498 (2013). https://doi.org/10.1021/jf401412s
  26. Kumar, P., Rubies, A., Companyo, R., Centrich, F.: Hydrophilic interaction chromatography for the analysis of aminoglycosides. J. Sep. Sci., 35, 498-504 (2011). https://doi.org/10.1002/jssc.201100860
  27. CODEX Alimentarius Commission, Guidelines on good laboratory practice in residue analysis. CAC/GL 40-1993 (2003).
  28. Ministry of Food and Drug Safety (MFDS), Guidelines on standard procedures for preparing analysis method (2016).