Isolation and Identification of Antimicrobial Active Substances from Rhodiola sachlinensis

홍경천(Rhodiola sachlinensis)에서 항균성 물질의 분리 및 동정

  • 심창주 (충남대학교 식품공학과) ;
  • 이규희 (충남대학교 식품공학과) ;
  • 정재홍 (충남대학교 식품공학과) ;
  • 이상덕 (충남대학교 식품공학과) ;
  • 김영호 (충남대학교 약학대학 약학부) ;
  • 오만진 (충남대학교 식품공학과)
  • Published : 2004.03.01

Abstract

The antimicrobial substances from Rhodiola sachalinensis were extncted, isolated and identified. The highest level of antimicrobial activity and its yield were obtained in methanol extract. The minimum inhibition concentrations of the methanol extract were 500 $\mu\textrm{g}$mL on agar plate and 100 $\mu\textrm{g}$mL in broth media for four gram positive and four gram negative microbials. The methanol extract was fractionated by n-hexane, chloroform, ethyl ether, ethyl acetate, and butanol, orderly. The separate was developed on the TLC plate with different solvent system ratio of chloroform and methanol. Nine substances were isolated from chloroform and methanol mixture(9:1, v/v). Among them, three isolates showed antimicrobial activity. Three substances separated by HPLC were identified by GC/MS(EI) spectrum and $^1$H, /sup13/C-NMR spectrum. They were gallic acid, (-)-epicatechin and kaempferol. The antimicrobial activities of each substances were shown gallic acid, (-)-epicatechin, kaempferol orderly.

홍경천의 천연 보존료로서의 이용성을 검토하기 위하여 각종 용매로 홍경천 추출물을 제조하고 극성에 따라 순차 분획, silica-gel chromatography, thin-layer chromatography, high performance liquid chromatography의 분리과정을 거쳐 순수 분리된 각 물질을 GC/MS(EI) spectrum, $^1$H-NMR, $^{13}$C-NMR spectrum, 을 이용하여 항균물질을 동정하여 다음과 같은 결과를 얻었다. 홍경천에 대해 acetone, ethyl acetate, ethanol, methanol, chloroform, water의 각 용매로 추출한 결과 항균성분의 추출 용매로는 methanol이 가장 우수하였다. 홍경천 methanol 추출물을 n-hexane, chloroform, ethyl ether, ethyl acetate, n-butyl alcohol로 분획한 결과 ethyl acetate와 n-butyl alcohol 분획에서 우수한 항균력을 나타내었다. 홍경천 methanol 추출물은 그람양성 4균주와 그람음성 4균주에 대해 고체 배지의 경우 대체로 500 $\mu\textrm{g}$mL에서 최소저해를 나타냈으며, 액체배지의 경우 100 $\mu\textrm{g}$mL 내외에서 최소저해를 나타내었다. Silica gel column chromatography에서 용매의 비율을 높여가며 용출한 결과 chloroform methanol(8:2, v/v)의 비율에서 항균력이 가장 높게 나타났다. 분리된 항균성분을 silica-gel TLC plate에 용매의 비율을 높여가며 전개시켜 chloroform methanol(9:1, v/v)에서 9개의 물질군을 얻을 수 있었으며, 그중 항균력이 확인된 3개의 물질군을 HPLC와 NMR spectrum을 분석한 결과 gallic acid, (-)-epicatechin 및 kaempferol로 동정되었다. gallic acid, (-)-epicatechit kaempferol의 항균물질 각각의 최소저해농도는 Staphylococcu aureus와 Listeria monocytogenes에 대해 gallic acid와 kaempferol이 500 $\mu\textrm{g}$mL(-)-epicatechin이 100 UgfmL에서 저해를 나타내었다.

Keywords

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