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Isolation and Identification of Antimicrobial Compounds against Helicobacter pylori from Rosemary (Rosmarinus officinalis L.) Extracts

로즈마리(Rosmarinus officinalis L.) 추출물로부터 Helicobacter pylori에 대한 항균물질 분리 및 동정

  • Yoon, So-Jung (Kyungjin Pharmacy) ;
  • Kim, Jin-Sung (School of Food Science, Kyungpook National University) ;
  • Jo, Bun-Sung (School of Food Science, Kyungpook National University) ;
  • Kim, Jeung-Hoan (School of Nano Materials Engineering, Kyungpook National University) ;
  • Lee, Sun-Ho (The Institute of Marine Biotechnology, Andong National University) ;
  • Ahn, Bong-Jeun (Department of Cosmeceutical Science, Daegu Hanny University) ;
  • Cho, Young-Je (School of Food Science & Biotechnology, Kyungpook National University)
  • 윤소정 (경진제약) ;
  • 김진성 (경북대학교 식품공학과) ;
  • 조분성 (경북대학교 식품공학과) ;
  • 김정환 (경북대학교 나노소재공학부) ;
  • 이선호 (안동대학교 해양바이오산업연구소) ;
  • 안봉전 (대구한의대학교 화장품약리학과) ;
  • 조영제 (경북대학교 식품공학부)
  • Received : 2010.03.22
  • Accepted : 2011.05.04
  • Published : 2011.09.30

Abstract

Rosmarinus officinalis extracts had a significant antimicrobial activity against Helicobacter pyori. Total phenolic contents and inhibition zone of rosemary extracts were estimated to be 25.7 mg/g and 14 mm at $200{\mu}g/mL$ of phenolic contents, respectively. The Sephadex LH-20 and MCI-gel CHP-20 column chromatographic separations for the phenolic extracts from R. officinalis leaves led to isolation of five acids, whose structures were determined as protocatechuic acid (A), coumaric acid (B), caffeic acid (C), chlorogenic acid (D), and rosmarinic acid (E), from interpretation of spectroscopic data including nagative fast atom bombardment (FAB)-mass, $^1H$-NMR, $^{13}C$-NMR, and IR. All isolated compounds were tested for antimicrobial activity against H. pyori. The purified single compound showed less antimicrobial activity against H. pylori than the mixed purified compounds, which generate A+B, A+E, C+D, C+E (each $200{\mu}g/disc$) excellent as large clear zone by synergy effect. These results indicate rosemary extracts are preventive agents against H. pyori.

로즈마리 추출물의 phenol 함량은 25.7 mg/g으로 매우 높게 나타났다. Helicobacter pylori 에 대한 항균효과를 측정한 결과, clear zone이 $100{sim}200{\mu}/disc$의 phenol 농도에서 각각 11, 12, 14 mm의 clear zone을 형성하여 추출물의 농도가 증가할수록 H. pylori 에 대한 저해효과가 농도 의존적으로 높아지는 것을 확인 할 수 있었다. Sephadex LH-20과 MCI-gel CHP-20 column을 이용하여 normal phase type인 $ethanol{\rightarrow}H_2O$와 reverse phase type인 $H_2O{\rightarrow}methanol$로 용액의 농도를 증가시키면서 용출한 결과 H. pylori에 대한 탁월한 저해활성을 나타낸 5종류(compound A~E)의 단일 물질로 분리하였다. 5가지 각각의 물질은 H. pylori에 대한 항균력이 미약하였으나, 2종류 이상의 다중 물질의 항균 시너지 효과에 의해 각각 10~16 mm의 clear zone을 형성하여 우수한 항균력을 나타내었으며, fast atom bombardment (FAB)-Mass, $^1H$-NMR, $^{13}C$-NMR과 IR spectrum을 사용하여 구조 동정한 결과 protocatechuic acid, coumaric acid, caffeic acid, chlorogenic acid 및 rosmarinic acid 인 것으로 확인되었다.

Keywords

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