Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Modification and Validation of an Analytical Method for Dieckol in Ecklonia Stolonifera Extract

곰피추출물의 지표성분 Dieckol의 분석법 개선 및 검증

  • Han, Xionggao (Department of Food Biotechnology and Environmental Science, Kangwon National University) ;
  • Choi, Sun-Il (Department of Food Biotechnology and Environmental Science, Kangwon National University) ;
  • Men, Xiao (Department of Food Biotechnology and Environmental Science, Kangwon National University) ;
  • Lee, Se-jeong (Department of Food Biotechnology and Environmental Science, Kangwon National University) ;
  • Oh, Geon (Department of Food Biotechnology and Environmental Science, Kangwon National University) ;
  • Jin, Heegu (Department of Food Science and Biotechnology, College of Life Science, CHA University) ;
  • Oh, Hyun-Ji (Department of Food Science and Biotechnology, College of Life Science, CHA University) ;
  • Kim, Eunjin (Naturalway Co. Ltd) ;
  • Kim, Jongwook (Naturalway Co. Ltd) ;
  • Lee, Boo-Yong (Department of Food Science and Biotechnology, College of Life Science, CHA University) ;
  • Lee, Ok-Hwan (Department of Food Biotechnology and Environmental Science, Kangwon National University)
  • 한웅호 (강원대학교 식품환경융합학과) ;
  • 최선일 (강원대학교 식품환경융합학과) ;
  • 문효 (강원대학교 식품환경융합학과) ;
  • 이세정 (강원대학교 식품환경융합학과) ;
  • 오건 (강원대학교 식품환경융합학과) ;
  • 진희구 (차의과학대학교 식품생명공학과) ;
  • 오현지 (차의과학대학교 식품생명공학과) ;
  • 김은진 ((주)네추럴웨이) ;
  • 김종욱 ((주)네추럴웨이) ;
  • 이부용 (차의과학대학교 식품생명공학과) ;
  • 이옥환 (강원대학교 식품환경융합학과)
  • Received : 2022.04.14
  • Accepted : 2022.06.10
  • Published : 2022.06.30
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Abstract

This study was to investigate an analytical method for determining dieckol content in Ecklonia stolonifera extract. According to the guidelines of International Conference on Harmonization. Method validation was performed by measuring the specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of dieckol using high-performance liquid chromatography-photodiode array. The results showed that the correlation coefficient of calibration curve (R2) for dieckol was 0.9997. The LOD and LOQ for dieckol were 0.18 and 0.56 ㎍/mL, respectively. The intra- and inter-day precision values of dieckol were approximately 1.58-4.39% and 1.37-4.64%, respectively. Moreover, intra- and inter-day accuracies of dieckol were approximately 96.91-102.33% and 98.41-105.71%, respectively. Thus, we successfully validated the analytical method for estimating dieckol content in E. stolonifera extract.

본 연구에서는 곰피추출물을 이용하여 건강기능식품 개발 시 원료의 표준화를 위하여 HPLC-PDA를 이용하여 지표성분 dieckol의 분석법 개선 및 검증을 실시하였다. 분석법의 유효성 검증은 ICH가이드라인에 근거하여 특이성, 직선성, 정확성, 정밀성, 검출한계 및 정량한계를 통해 신뢰성 및 타당성을 검증하였다. 지표성분 dieckol 분석을 위하여 0.1% formic acid가 첨가된 water와 acetonitrile을 이동상으로 사용하였고 컬럼은 Capcell pak C18을 사용하여 HPLC를 이용한 분석방법에서 표준용액과 곰피추출물의 지표성분 dieckol 피크 머무름 시간 일치하고 다른 물질의 간섭을 받지 않는 것을 확인하였으며, PDA spectrum이 일치하므로 특이성을 확인하였다. Dieckol의 검량선의 상관계수(R2)은 1에 가까운 높은 상관계수 값 0.9997로 높은 직선성을 확인할 수 있었으며 분석에 적합함을 알 수 있었다. 농도를 알고 있는 곰피추출물에 dieckol 표준용액을 최종 농도가 각각 저농도(2.5 ㎍/mL), 중간농도(5 ㎍/mL), 고농도(10 ㎍/mL)가 되도록 첨가하여 정밀성 및 정확성을 계산하였다. 정밀성은 일간, 일내 정밀성으로 확인하였으며, intra-day 및 inter-day 분석에서 RSD는 각각 1.58-4.39%, 1.37-4.64% 수준으로 5% 이하의 우수한 정밀성을 보였다. 정확성 측정결과 intra-day 및 inter-day 분석에서 회수율은 각각 96.91-102.33% 및 98.41-105.71%로 우수한 정확성을 나타내었다. 검출한계(LOD)와 정량한계(LOQ)는 0.18 ㎍/mL, 0.56 ㎍/mL로 각각 확인되였으며 낮은 농도에서도 검출이 가능함을 보였다. 확립된 분석법은 특이성, 직선성, 정밀성, 정확성, 검출한계 및 정량한계에 대한 분석법 검증결과가 모두 우수한 분석방법임을 증명하였다. 또한 검증된 분석법을 이용하여 곰피추출물 중 함유한 dieckol 함량을 분석한 결과 27.39±0.50 mg/dry weight g로 분석되었다. 본 연구는 HPLC-PDA를 이용한 곰피추출물의 지표성분인 dieckol의 분석방법을 개선 및 유효성 검증을 진행하여 과학적으로 신뢰성이 있는 적합한 분석방법임이 검증되었다.

Keywords

Acknowledgement

이 논문은 2021년 해양수산부 재원으로 해양수산과학기술진흥원(곰피추출물을 이용한 체지방 감소 개별인정형 소재 개발 및 제품 상용화), 2021년도 교육부의 재원(NRF-2021R1A6A1A03044242)과 한국연구재단의 4단계 두뇌한국21 사업(4단계 BK21 사업)(4299990913942)의 지원으로 수행되었으며 이에 감사드립니다.

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