Development of Rapid Analytical Method of Forbidden Medicines in Dietary Supplements Using LC-ESI-Tandem MS

LC-ESI-tandem MS를 이용한 기능성표방식품 중 부정유해물질 신속검사체계 개발

  • Kim, Hee-Yun (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration) ;
  • Jang, Young-Mi (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration) ;
  • Joo, Hyun-Jin (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration) ;
  • Jung, Young-Hyun (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration) ;
  • Lee, Myoung-Sook (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration) ;
  • Park, Jong-Seok (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration) ;
  • Lee, Kwang-Ho (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration) ;
  • Lee, Hwa-Mi (Hazard Substances Analysis Team, Center for Food and Drug Inspection, Gyeongin Regional Korea Food and Drug Administration)
  • 김희연 (경인지방식품의약품안전청 유해물질분석팀) ;
  • 장영미 (경인지방식품의약품안전청 유해물질분석팀) ;
  • 주현진 (경인지방식품의약품안전청 유해물질분석팀) ;
  • 정용현 (경인지방식품의약품안전청 유해물질분석팀) ;
  • 이명숙 (경인지방식품의약품안전청 유해물질분석팀) ;
  • 박종석 (경인지방식품의약품안전청 유해물질분석팀) ;
  • 이광호 (경인지방식품의약품안전청 유해물질분석팀) ;
  • 이화미 (경인지방식품의약품안전청 유해물질분석팀)
  • Published : 2007.08.31

Abstract

A high-performance liquid chromatography-electrospray ionization (HPLC-ESI) tandem MS was developed for the rapid and simultaneous determination of forbidden medicines in dietary supplements. Thirteen medicinal components such as PDE-5 inhibitors and their analogues, and the newly identified dimethylsildenafil and xanthoanthrafil, were included in this study. After tentative standardization of molecular ions in both polarities using thirteen references on the mass spectrometer, with ESI-continuous infusion via the syringe pump method, the relative intensity of the ions present in the resulting spectra was quantitatively compared. From the results, the ion mode was selected depending on each reference's characteristics. A HPLC method coupled with the ESI mode was developed considering the matrix effect and interference depending on the type of sample. The validation test of the developed method was followed by carrying out precision, accuracy, recovery, sensitivity and linearity, etc. The method showed sufficiently high sensitivity, reproducibility, and specificity, and produced 4 times faster results when compared with the existing HPLC/UV method for the determination of forbidden compounds in dietary supplements.

부정유해물질 총 13종에 대한 신속하고 고감도의 LC-ESI-MS-MS 동시분석 방법을 개발하였으며 바데나필을 포함한 11종은 ESI positive 모드에서 타다라필을 포함한 2종은 ESI negative 모드에서 검출하는 방법으로서 시료 전처리는 간단히 메탄올 추출법을 사용하였다. 기능성표방식품 중 부정유해물질의 확인은 한번 시료를 주입함으로써 15분 이내에 13종의 분석이 가능하고 크로마토그램의 분리는 아세토니트릴과 10mM ammonium formate가 들어있는 탈이온수를 이용한(pH 7.0) 기울기 용매 조건으로 수행하였다. 확립한 13종에 대한 부정유해물질 분석방법은 검출 한계(LOD)는 0.1-5 ng/mL이고, 정량한계(LOQ)는 0.1-10 ng/mL으로서 평균 상관계수$(r^2)$는 0.9853로서 ppb 수준에서 정량성을 가지며 회수율은 87.5-98.5%, 변동계수는 15% 이하임을 확인할 수 있었다. 또한 확립한 시험방법 LC/MS/MS를 이용하여 147건의 기능성표방식품 중의 부정유해물질의 검증을 실시한 결과, 유해물질의 검출이 나타타지 않음을 알 수 있었다. 기능성표방식품 중의 실데나필과 그 유사물질을 포함한 13종의 부정유해물질에 대한 스크리닝 방법으로 MRM 모드를 이용한 LC-ESI-MS-MS 방법을 개발하였으며, 이는 유해물질에 대한 고성능액체크로마토그래피/자외선흡광광도법의 선택성등의 제한성을 극복한 부정유해 물질의 스크리닝에 신속하고 미량까지 검출 가능한 가치 있는 방법임을 확인할 수 있었다.

Keywords

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