Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Development and Validation of Analytical Method for Wogonin, Quercetin, and Quercetin-3-O-glucuronide in Extracts of Nelumbo nucifera, Morus alba L., and Raphanus sativus Mixture

연잎, 상엽, 건조 무 혼합 추출물의 지표성분 wogonin, quercetin 및 quercetin-3-O-glucuronide의 분석법 개발 및 검증

  • Jang, Gill-Woong (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Park, Eun-Young (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Choi, Seung-Hyun (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Choi, Sun-il (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Cho, Bong-Yeon (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Sim, Wan-Sup (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Han, Xinggao (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Cho, Hyun-Duk (Haram Co. Ltd.) ;
  • Lee, Ok-Hwan (Department of Food Science and Biotechnology, Kangwon National University)
  • 장길웅 (강원대학교 식품생명공학과) ;
  • 박은영 (강원대학교 식품생명공학과) ;
  • 최승현 (강원대학교 식품생명공학과) ;
  • 최선일 (강원대학교 식품생명공학과) ;
  • 조봉연 (강원대학교 식품생명공학과) ;
  • 심완섭 (강원대학교 식품생명공학과) ;
  • 한웅호 (강원대학교 식품생명공학과) ;
  • 조현덕 (하람) ;
  • 이옥환 (강원대학교 식품생명공학과)
  • Received : 2018.05.30
  • Accepted : 2018.07.02
  • Published : 2018.08.30
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Abstract

The aim of this study was to develop and validate an analytical method for determining the presence of wogonin, quercetin, and quercetin-3-O-glucuronide in extracts of Nelumbo nucifera, Morus alba L., and Raphanus sativus mixtures. We evaluated the specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of analytical methods for wogonin, quercetin, and quercetin-3-O-glucuronide using high performance liquid chromatography. Our result showed that the correlation coefficients of the calibration curve for wogonin, quercetin, and quercetin-3-O-glucuronide were 0.9999. The LOD for wogonin, quercetin, and quercetin-3-O-glucuronide ranged from 0.09 to 0.16 and those for the LOQ ranged from 0.26 to $0.48{\mu}g/mL$. The inter-day and intra-day precision values of wogonin, quercetin, and quercetin-3-O-glucuronide ranged from 0.74 to 1.87 and from 0.28 to 1.12%, respectively. The inter-day and intra-day accuracies were 99.96~115.88% and 99.73~114.81%, respectively. Therefore, the analytical method was validated for the detection of wogonin, quercetin, and quercetin-3-O-glucuronide in extracts of Nelumbo nucifera, Morus alba L., and Raphanus sativus mixtures.

본 연구는 연잎, 상엽, 건조 무 혼합 추출물의 wogonin, quercetin, quercetin-3-O-glucuronide의 함량 분석 및 해당 표준물질들의 동시 분석법의 개발 및 검증을 실시하였다. 분석법은 특이성, 직선성, 정확성, 정밀성, 검출한계 및 정량한계를 통하여 분석법의 신뢰성을 검증했다. HPLC를 이용한 분석방법에서 표준용액의 머무름 시간과 연잎, 상엽, 건조 무 혼합 추출물 중 quercetin-3-O-glucuronide의 머무름 시간이 일치하였으며 동일한 spectrum을 나타내는 것을 확인해 분석법의 특이성을 검증하였다. Wogonin, quercetin 및 quercetin-3-O-glucuronide의 검량선은 상관계수 값이 wogonin, quercetin 및 quercetin-3-O-glucuronide 모두 0.9999로 우수한 직선성을 보여 분석에 적합함을 알 수 있었고 정밀성과 정확성은 농도를 아는 시료에 인위적으로 저농도, 중농도, 고농도의 표준물질을 첨가해 계산하였다. Wogonin, quercetin, quercetin-3-O-glucuronide의 정밀성은 일간 정밀성, 일내 정밀성으로 확인했으며 세 가지 성분들의 일간 정밀성은 각각 0.76~1.24%, 0.90~1.40%, 0.74~1.87% 수준으로 확인되었고, 일내 정밀성은 0.12~1.06%, 0.28~1.12%, 0.52~0.92%로 5% 이하의 우수한 정밀성을 보였다. 정확성의 측정 결과 wogonin, quercetin 및 quercetin-3-O-glucuronide의 일간 정확성은 99.99~107.97%, 99.96~115.88%, 99.73~111.91%를 나타냈고 일내 정확성은 100.36~108.88%, 99.77~114.81%, 99.91~112.44%로 우수한 수준의 정확성을 보였다. Wogonin, quercetin 및 quercetin-3-O-glucuronide의 검출한계는 각각 $0.16{\mu}g/mL$, $0.09{\mu}g/mL$, $0.15{\mu}g/mL$로 측정됐으며 정량한계는 각각 $0.48{\mu}g/mL$, $0.26{\mu}g/mL$, $0.45{\mu}g/mL$로 나타나 저농도에서도 검출이 가능함을 확인할 수 있었다. 분석법 검증결과, 확립된 분석법은 특이성, 직선성, 정밀성, 정확성, 검출한계 및 정량한계가 모두 우수한 분석법임이 검증됐으며, 검증된 분석법을 이용해 연잎, 상엽, 건조 무 혼합 추출물의 wogonin, quercein, quercetin-3-O-glucuronide의 함량을 분석한 결과 wogonin과 quercetin은 검출이 되지 않았지만 quercetin-3-O-glucuronide는 함량을 가지고 있는 것으로 확인됐다. 본 연구 결과 에탄올로 추출한 11번(EM11) 샘플에서 quercetin-3-O-glucuronide의 함량이 가장 높게 나왔으며 열수와 에탄올로 추출한 샘플들이 함유하고 있는 quercetin-3-Oglucuronide의 함량은 불검출~50.95 mg/dry weight g 수준으로 함유되어 있는 결과를 보였다.

Keywords

References

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