Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Improvement of an Simultaneous Determination for Clenbuterol and Ractopamine in Livestock Products using LC-MS/MS

LC-MS/MS를 이용한 축산물 중 clenbuterol과 ractopamine의 동시 분석법 개선

  • Cho, Yoon-Jae (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Chae, Young-Sik (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Kim, Jae-Young (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Kim, Hyung-Soo (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Kang, Ilhyun (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Do, Jung-Ah (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Oh, Jae-Ho (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Kwon, Kisung (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Chang, Moon-Ik (Food Chemical Residues Division, Department of Food Safety Evaluation, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration)
  • 조윤제 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 채영식 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 김재영 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 김형수 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 강일현 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 도정아 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 오재호 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 권기성 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과) ;
  • 장문익 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 화학물질과)
  • Received : 2012.11.29
  • Accepted : 2013.01.21
  • Published : 2013.02.28
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Abstract

Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

동물용의약품은 2007년부터 급격한 잔류허용기준 신설에 따라 많은 수의 분석법도 함께 신설하였으며, 국제식품규격위원회(CODEX), EU 등에서 동물용의약품에 대한 기준이 국제적으로 엄격해지고 있어, 낮은 농도의 정량한계 및 재현성이 높은 분석법이 요구되어지고 있다. 하지만 국내 식품공전에서의 클렌부테롤 및 락토파민 분석법은 각각 개별 분석법으로 나뉘어져 있고, 시간적 및 경제적으로 손실이 있을 뿐 아니라 추출 효율 및 재현성이 낮아 분석에 어려움이 있다. 따라서 본 연구는 물리화학적 특성이 유사한 ${\beta}$-agonist계 동물용의약품인 클렌부테롤 및 락토파민의 기존 개별 분석법을 동시 분석법으로 개선하고 검사 효율성을 증대시키고자 하였다. 분석에 사용된 검체는 소와 돼지의 근육을 이용하였다. 검체에 내부표준물질인 클렌부테롤-$d_9$과 락토파민-$d_3$을 각각 첨가하고 ${\beta}$-글루쿠로니다제/아릴설파타제 효소를 사용하여 가수분해한 후 에틸아세테이트로 추출하였다, 추출액을 농축한 후 헥산과 메탄올을 포화시킨 용매를 적용하여 지방 제거과정을 거친 뒤 MIP 카트리지로 정제한 후 액체크로마토그래피-질량분석기(LC-MS/MS)에 주입하였다. 기기분석은 ESI(Electro-Spray Ionization) 및 positive MRM(Multiple Reaction Monitoring) 모드로 하였고, 검증은 CODEX 가이드라인 규정에 따라 실시하였다. 그 결과, 클렌부테롤과 락토파민의 LOQ는 각각 0.2 및 0.5 ${\mu}g/kg$ 수준이었고, 평균회수율은 각각 104.2-113.5% 및 107.6-118.1%로 나타났다. 또한, 분석오차는 각각 2.8-10.5% 및 1.6-5.2%로 CODEX 가이드라인 규정에 만족하는 수준이었다. 따라서 개선된 동시 분석법은 잔류동물용의약품의 분석에 있어 보다 신속하고 경제적인 분석 및 모니터링에 적용 가능할 것으로 기대된다.

Keywords

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