Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
권호 표지

Isolation an Identification of the Active Compounds from Green Mustard Leaves against Oxidative Stress in Bovine Brain Tissue

청겨자채 추출물의 뇌조직 내 Oxidative Stree 억제활성물질의 동정

  • 이근종 (연세대학교 생물산업소재연구센터) ;
  • 김미리 (충남대학교 식품영양학과) ;
  • 심재석 (연세대학교 생물소재공학과) ;
  • 황재관 (충남대학교 식품영양학과)
  • Published : 2001.10.01
Download PDF
⟨ Previous Next ⟩

Abstract

Green mustard leves were found to effectively prevent lipid peroxidation of bovine-brain tissue by ascor-bate/Fe system, The 50% methanol extracts mustard leaves were separated into four solvent faction using n-hexane,. EtOAc, n-BiOH and water. Then n-BiOH fraction exclusively exhibited the antioxidative activities at concentration above 100 $\mu\textrm{g}$/mL/ The n-BuOH fraction was further isolated to a single compound using TLC analysis and silica gel chromatography. The active antiodidative compounds were identified as sinapic acid methyl ester and ferulic acid methyl ester by $^{1}$H-NMR and $^{13}$ C-NMR, The sinapic acid methyl ester and ferulic acid methyl ester were prepared by methylating of sinapic acid and ferulic acid with diazomethane. The results strongly suggested that sinapic acid and ferulic acid could be emplyed as a potential antioxiative agents for preventing the bovine brain lipid peroxidation. lipid peroxidation.

마우스 뇌조직에서 지질과산화를 막아주는 것으로 알려진 청겨자채에 대해서 50%MeOH 추출물(2mg/mL)을 사용하여 소 뇌조직의 산화적스트레스를 유발한 ascorbate/Fe$^{3+}$ system에서 항산화 활성을 조사하였다. 청겨자채 추출물의 용매순차추출 후 TBARS 활성을 각각 용매 분획(100)$\mu\textrm{g}$/mL)별로 확인한 결과 각각의 용매 분획 중 n-BiOH 분획 (49%)이 가장 높았고, EtOAc(40%), H$_{2}$O(9%) 및 n-hexane (0%) 순이었다. n-BuOH 분획을 silica gel column chroma-tography를 반복하였으며, CE, TLC를 통해서 sinapic acid와 ferulic acid를 확인하였다. CE를 통하여 그 함량을 정량한 결과, 건조중량(g)당 sinapic acid는 24.3 ppm 이었고 ferulic acid는 42.8 ppm으로 나타났다. 최종 정제활성물질을 $^{1}$J-NMR(CDCL$_{3}$)와 $^{13}$C-NMR(CDCL$_{3}$)로 확인하였으며 sinapic acid methyl ester, ferulic acid acid와 ferulic acid와 RF치가 일치하는지 여부를 TLC상에서 비교하였다. 동일한 50$\mu\textrm{g}$/mL 농도에서 sinapic acid methyl ester는 40%, ferulic acid methyl ester 는 35%의 항산화 활성을 나타내었으며 이것은 시판되는 표준물질은 sinapic acid(22%)와 ferulic acid (18%)에 비해 약간 높게 나타났다. 이것은이 화합물에 라디칼을 소거하는 작용기와 관계가 있는 것으로 여겨진다. 청겨자채 추출물 중에도 항산화 활성이 비교적 널리 분포된 것으로 보아 이 성분들은 체내에서 단독으로 또는 상호작용으로 항산화 작용을 나타낼 것으로 예상된다.

Keywords

References

  1. Oxidative stress, antioxidants, aging and disease: Oxidative stess and aging Cutler, R. G.;Packer, L.;Bertram, J.;Mori, A.;Packer, L.(ed.)
  2. Biochem. Biophys. Res. Commun. v.78 Hydroxyl radical production involved in lipidperoxidation of rat microsomes Lai, C. S.;Pertte, L. H.
  3. J. Biol. Chem. v.248 Evidence that peroxidation of lysosomal membranes is intiated by hydroxyl free radicals producedflavin enzyme activity Fong, K. L.;McCay, P. B.;Poyer, J. L.;Keele, B. B.;Misra, H.
  4. Cell. Biol. Int.Rep. v.2 Biological mechanisms accounting for the toxic action of oxygen onliving organisms: The key role of superoxide dismutase Halliweel, B.
  5. Biochem. J. v.224 Reactivity of hydroxyl and hydroxyl-like radicals discriminated by release of thiobarbituric acid-reactive material from deoxy sugars, nucleosidesandbenzoate Gutteridge, J. M.
  6. Cell Biophysics. v.14 Scavenging effect of extracts of green tea natural antioxidants on active oxygen radicals Zhao, B. L.;Li, X. J.;He, R. G.;Chens, S. J.;Win, W. J.
  7. Free Radic. Res. Commun. v.16 Essential oil phenyl propanoids: Useful as □OH scavengers? Taira, J.;Ikemoto, T.;Yoneya, T.;Hagi, A.;Murakami, A.;Makino, K.
  8. Biosci. Biotech. Biochem. v.61 Hydroxyl radical-scavenging effects ofspices and scavengers from brown mustard (Brassica nigra) Chung, S. K.;Osawa, T.Kawakishi, S.
  9. Food Sci. Biotechnol. v.7 Hydroxyl radical scavengers from white mustard (Sinapis alba) Chung, S. K.;Osawa, T.
  10. Kor. J. Pharmacogn v.18 Studies on anti-oxidant componet of mustard leaf and seed Han, Y. B.;Kim, M. R.;Han, B. H.;Han, Y. N.
  11. Cancer Res. v.45 Chemoprevention of cancer Watenberg, L. W.
  12. Ph.D. Dissertation, Chunam Nasional University Protetive effect against oxidative stress in mouse brain tissue and induction of hepatic glutathione S-transferase vegetable extracts Lee, K. J.
  13. J. Korean Sco. Food Nutr. v.24 Antimicrobial substances of distilled components from mustard seed Shim, K. H.;Seo, K. I.;Kang, K. S.;Moon, J. S.;Kim, H. S.
  14. J. Korean Soc. Dietary Culture v.8 Purification and charicterization of myrosinase in Dolsan leaf mustard (Brassica juncea) and changes in myrisinase acitvity during fermentation of leaf mustard kimchi Park, J. R.;Park, S. K.;Cho, Y. S.;Chun, S. S.
  15. J. Assoc. Official Anal. Chemists v.66 Analytical methodology for determining glucosinolate composition and content McGregor, D. J.;Mullin, W. J.;Fenwick, G. R.
  16. Oil-Chemistry v.26 Antioxidant properties of spices Saito, Y.
  17. Food Res. International v.31 Flavour and pharmaceutical properties of the volatile sulphur compounds of wasabi (Wasabia japonica) Depree, J. A.;Howard, T. M.Savage, G. P.
  18. Symposium of Korean Society of Food Science and Nutrition, Se Inhibitory effect of wasabi on the urease active of Helicobacter pylori Shin, I. S.;Masuda, H.;Kinae, N.
  19. Nat. Prod. Sci. v.64 Study of flavonoid compound from mustard leaf (Brassica juncea) Kim, J. E.;Choi, J. S.
  20. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding Bradford, M. M.
  21. Lipids v.8 Damage to microsomal membrane by lipidperoxidation Bidlack, W. T.;Tappel, A. L.
  22. Org. Syn. v.36 Preparation of diazomethane Deboer, T. J.;Backer, H. J.
  23. Tables of spectral data for structure determination of organic compounds(2nd ed.) Pretsh, E.;Seibl, J.;Simon, W.;Clere, T.
  24. Biosci. Biotech. Biochem. v.56 Comparison of the antioxidative activity of some acid-phenols: Structure-activity relationship Cuvelier, M. E.;Richard, H.;Berset, C.