Journal of the Society of Cosmetic Scientists of Korea (대한화장품학회지)

Society of Cosmetic Scientists of Korea (대한화장품학회)
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Antioxidative Activity and Component Analysis of Quercus glauca Leaf Extracts

종가시나무 잎 추출물의 항산화 활성, 성분 분석

  • Yang, Hee-Jung (Department of Fine Chemistry, College of Nature and Life Science, Seoul National University of Technology) ;
  • Ahn, You-Jin (Department of Fine Chemistry, College of Nature and Life Science, Seoul National University of Technology) ;
  • Kim, Jae-Hyun (Department of Fine Chemistry, College of Nature and Life Science, Seoul National University of Technology) ;
  • Park, Soo-Nam (Department of Fine Chemistry, College of Nature and Life Science, Seoul National University of Technology)
  • 양희정 (서울산업대학교 자연생명과학대학 정밀화학과) ;
  • 안유진 (서울산업대학교 자연생명과학대학 정밀화학과) ;
  • 김재현 (서울산업대학교 자연생명과학대학 정밀화학과) ;
  • 박수남 (서울산업대학교 자연생명과학대학 정밀화학과)
  • Published : 2008.09.30
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Abstract

In this study, the antioxidative effects, inhibitory effects on elastase, and components of Quercus glauca extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ of extract I fractions of Quercus glauca leaf was in the order: 50% ethanol extract $(12.45{\mu}g/mL)$ < ethyl acetate fraction $(10.47{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(8.57{\mu}g/mL)$. Reactive oxygen species (ROS) scavenging activities $(OSC_{50})$ of some Quercus glauca leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was 50% ethanol extract $(OSC_{50},\;4.2{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(1.58{\mu}ug/mL)$ < ethyl acetate fraction $(0.66{\mu}g/mL)$. Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract / fractions of Quercus glauca leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Quercus glauca leaf extracts suppressed photohemolysis in a dose dependent manner, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect $({\tau}_{50}$, 398.67 min at $50{\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Quercus glauca leaf extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (360 nm) as well. Two components were identified as quercetin (55.77%), and kaempferol (44.23 %). TLC chromatogram of ethyl acetate fraction of Quercus glauca leaf extracts revealed 6 bands $(QG1{\sim}QG6)$, Among them, isoquercitrin (QG3), hyperin (QG4), and rutin (QG6) were identified. The inhibitory effect of aglycone fraction on tyrosinase $(IC_{50},\;73.5{\mu}g/mL)$ and elastase $(IC_{50},\;16.2{\mu}g/mL)$ was high. These results indicate that extract / fractions of Quercus glauca can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Quercus glauca leaf extract and inhibitory activity on tyeisinase and elastase of the aglycone fraction could be applicable to new functional cosmetics.

본 연구에서는 종가시나무 잎 추출물의 항산화, 성분 분석 및 elastase 저해 효과에 관한 조사를 수행하였다. 추출물의 free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) 소거환성 $(FSC_{50})$은 50% 에탄을 추출물$(12.45{\mu}g/mL)$ < ethyl acetate 분획 (10.47) < 당을 제거시킨 플라보노이드 aglycone 분획 (8.57) 순으로 증가하였다. Luminol-의존성 화학발광법을 이용한 $Fe^{3+}-EDTA/H_2O_2$ 계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 종가시나무 잎 추출물의 총항산화능은 50% 에탄올 추출물$(OSC_{50},\;4.2{\mu}g/mL)$ < aglycone 분획(1.58) < ethyl acetate 분획 (0.66) 순으로, ethyl acetate 분획에서 가장 큰 활성을 나타내었다. 종가시나무 잎 추출물에 대하여 rose-bengal로 증감된 사람적혈구의 광용혈에 대한 억제 효과를 측정하였다. 종가시나무 잎 추출물의 경우 농도 의존적$(1{\sim}50{\mu}g/mL)$으로 광용혈을 억제하였다. 특히 당을 제거시킨 aglycone 분획은 $50{\mu}g/mL$ 농도에서 ${\tau}_{50}$ 398.67 min으로 매우 큰 세포보호 효과를 나타내었다. 종가시나무 잎 추출물 중 ethyl acetate 분획의 당 제거 반응 후 얻어진 aglycone 분획은 TLC에서 2개의 띠고 분리되었으며, HPLC 실험 (360 nm)에서도 2개의 피이크로 분리되었다. 분리된 2가지 성분은 quercetin 및 kaempferol이었으며, 그들의 성분비는 각각 55.77%, 44.23%로 quercetin의 함량이 큰 것으로 나타났다. 종가시나무 잎 추출물의 ethyl acetate분획의 TLC 크로마토그램은 6개의 띠$(QG1{\sim}QG6)$로 분리되었고. QG 3는 isoquercitrin, QC 4는 hyperin, QG 6는 rutin으로 확인되었다. Aglycone 분획은 tyrosinase 저해활성 $(IC_{50})$이 $73.5{\mu}g/mL$이고, elastase 저해환성 $(IC_{50})$은 $16.2{\mu}g/mL$로 활성을 나타내었다. 이상의 결과들은 종가시나무 잎 추출물이 $^1O_2$ 혹은 다른 ROS를 소광시키거나 소거함으로써 그리고 ROS에 대항하여 세포막을 보호함으로써 생체계, 특히 태양 자외선에 노출된 피부에서 항산화제로서 작용할 수 있음을 가리키며, 종가시나무 잎 성분 분석과 ethyl acetate 분획의 당 제거 실험 후 얻어진 aglycone분획의 tyrosinase, elastase 저해활성으로부터 기능성 화장품원료로서 응용 가능성이 있음을 시사한다.

Keywords

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