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 and Antioxidant Effect for Berberine and Palmatine in P.amurense

황백의 지표성분 berberine과 palmatine의 분석법 개발과 검증 및 항산화 효능 평가

  • Jang, Gill-Woong (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Choi, Sun-Il (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Han, Xionggao (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Men, Xiao (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Kwon, Hee-Yeon (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Choi, Ye-Eun (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Park, Byung-Woo (CombiMed Co. Ltd.) ;
  • Kim, Jeong-Jin (CombiMed Co. Ltd.) ;
  • Lee, Ok-Hwan (Department of Food Science and Biotechnology, Kangwon National University)
  • Received : 2020.10.30
  • Accepted : 2020.11.18
  • Published : 2020.12.30
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Abstract

The aim of this study was to develop and validate a simultaneous analytical method for berberine and palmatine, which are representative substances of Phellodendron amurense, and to evaluate the antioxidant activity. We evaluated the specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of analytical methods for berberine and palmatine using high-performance liquid chromatography. Our result showed that the correlation coefficients of the calibration curve for berberine and palmatine exhibited 0.9999. The LODs for berberine and palmatine were 0.32 to 0.35 µg/mL and the LOQs were 0.97 to 1.06 µg/mL, respectively. The inter-day and intra-day precision values for berberine and palmatine were from 0.12 to 1.93 and 0.19 to 2.89%, respectively. The inter-day and intra-day accuracies were 98.43-101.45% and 92.39-100.60%, respectively. In addition, the simultaneous analytical method was validated for the detection of berberine and palmatine. Moreover, we conducted FRAP and NaNO2 scavenging activity assays to measure the antioxidant activities of berberine and palmatine, and both showed antioxidant activity. These results suggest that P.amurense could be a potential natural resource for antioxidant activity and that the efficacy can be confirmed by investigating the content of the berberine and palmatine.

본 연구의 목적은 예로부터 약재로 사용되어 온 황백의 대표적인 지표물질 berberine 및 palmatine의 동시 분석법을 개발 및 검증하고 지표물질들의 항산화 효능을 평가하여 해당 물질들에 대한 기초자료를 제공하고자 하였다. 표준물질들의 동시 분석법은 특이성, 직선성, 정확성, 정밀성, 검출한계 및 정량한계를 측정하여 신뢰성을 검증하였으며, 지표물질들의 항산화 활성은 FRAP 활성, reducing power 및 아질산염 소거능 실험으로 측정하였다. HPLC-PDA를 이용하여 berberine과 palmatine의 혼합용액을 분석하였을 때, 두 물질이 간섭 없이 선택적으로 분리되었음을 확인하였다. Berberine 및 palmatine의 검량선의 상관계수 값은 모두 0.9999로 우수한 직선성을 보여 분석에 적합함을 알 수 있었고 정확성과 정밀성은 농도를 알고 있는 황백 시료에 인위적으로 저농도, 중농도, 고농도의 표준물질 혼합용액을 첨가하여 계산하였다. 정밀성과 정확성은 일간실험 및 일내실험으로 확인하였다. Berberine의 일간 및 일내 정밀성은 1.27-1.93%, 0.12-0.56%의 수준으로 나타났고, palmatine의 일간 및 일내 정밀성은 0.19-2.89%, 0.36-1.52%로 % 이하의 우수한 정밀성을 나타냈으며, berberine의 일간 및 일내 정확성은 98.43-100.73%, 99.56-101.45%로 나타났고 palmatine의 일간 및 일내 정확성은 93.20-100.60%, 92.39-97.10%로 나타났다. Berberine과 palmatine의 검출한계는 각각 0.32 µg/mL, 0.35 µg/mL로 확인되었고 정량한계는 0.97 µg/mL, 1.06 µg/mL로 확인되었다. 분석법 검증결과, 확립된 분석법은 특이성, 직선성, 정밀성, 정확성, 검출한계 및 정량한계가 모두 우수한 분석법임을 검증할 수 있었다. 항산화 활성은 FRAP 활성, reducing power 및 berberine과 palmatine 모두 농도 의존적으로 활성이 증가하는 것을 확인할 수 있었다. 이상의 결과를 종합하여 볼 때 황백은 추후 항산화에 도움을 주는 소재로의 적용 및 항염증 효과로 인한 피부 기능개선 제품의 소재로서 활용도가 넓을 것으로 사료되며, 황백 내에 존재하는 항산화물질인 berberine 및 palmatine의 동시분석법을 통하여 성분의 함량을 분석하여 효능을 확인할 수 있을 것으로 사료된다.

Keywords

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