Components and Their Antioxidative Activities of Methanol Extracts from Sarcocarp and Seed of Zizyphus jujuba var. inermis Rehder

생대추(풋대추) 과육 및 씨 추출물의 항산화 효과 및 항산화 성분

  • Yu, Mi-Hee (Department of Food Science and Technology, Keimyung University) ;
  • Im, Hyo-Gwon (Department of Food Science and Technology, Keimyung University) ;
  • Lee, Hyo-Jung (Department of Food Science and Technology, Keimyung University) ;
  • Ji, Young-Ju (Department of Food Science and Technology, Keimyung University) ;
  • Lee, In-Seon (The Center for Traditional Microorganism Resources, Keimyung University)
  • 유미희 (계명대학교 식품가공학) ;
  • 임효권 (계명대학교 식품가공학) ;
  • 이효정 (계명대학교 식품가공학) ;
  • 지영주 (계명대학교 식품가공학) ;
  • 이인선 (계명대학교 전통미생물자원개발 및 산업화연구(TMR) 센터)
  • Published : 2006.02.01

Abstract

Antioxidant activities of methanol extracts from sarcocarp and seed of Zizyphus jujuba var. inermis Rehder were investigated in vitro. Contents of total polyphenols in methanol extracts from sarcocarp and seed were 98.83, $138.99\;{\mu}g/mg$, respectively. Radical-scavenging activities of methanol extracts were examined using ${\alpha},{\alpha}-diphenyl-{\beta}-picrylhydrazyl$ and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals, and hydrogen peroxide assay. Inhibition effects of methanol extracts on peroxidation of linoleic acid were examined by ferric thiocyanate and thiobarbituric acid methods. Both sarcocarp and seed of Zizyphus jujuba var. inermis Rehder showed relatively high antioxidant activities in various systems.

본 실험에서는 풋대추의 유용자원으로의 이용가능성을 알아보기 위해, 풋대추의 과육과 씨를 각각 분리한 후 추출물을 제조하여 이들 각각의 총 폴리페놀 및 총 플라보노이드의 함량 측정하고 천연항산화제 및 합성항산화제로 널리 알려진 ascorbic acid와 BHA와의 비교측정으로 생대추의 항산화 활성을 검색해 보았다. 먼저 대추의 과육 및 씨의 메탄올 추출물에 존재하는 총 폴리페놀 및 플라보노이드 함량을 측정한 결과, 대추과육 추출물의 총 폴리페놀 함량은 $98.83\;{\mu}g/mg$, 대추씨 추출물은 $138.99\;{\mu}g/mg$으로 대추과육 보다는 대추씨에서 높은 폴리페놀 함량을 보였다. 총 플라보노이드 함량은 대추과육과 대추씨 추출물이 각각 35.56 및 $131.48\;{\mu}g/mg$으로 대추씨에서 플라보노이드 함량이 높게 나타났다. 각 시료의 DPPH 소거 활성을 농도별로 측정하여 비교한 결과, $10\;{\mu}g/mL$의 농도에서 대추과육이 92%, 대추씨가 95%의 소거능을 보였고, BHA와 ascorbic acid$(10\;{\mu}g/mL)$에서 96% 정도의 항산화능을 보였다. 또한 $ABTS{\cdot}^+$ 소거활성을 Trolox, BHA, ascorbic acid와 비교 측정한 결과, $ABTS{\cdot}^+$ 소거활성법에서 표준물질로 사용되는 Trolox는 $15\;{\mu}M$에서 63% 정도의 소거활성능을 보였고, 추출물은 $100\;{\mu}g/mL$에서 98-99% 정도의 $ABTS{\cdot}^+$의 높은 소거활성을 보였으며, BHA와 ascorbic acid는 $10\;{\mu}g/mL$에서 99% 이상의 소거활성능을 보였다. Hydrogen peroxide 소거활성을 측정한 결과 같은 농도에서 DPPH radical이나 ABTS radical 소거활성보다 다소 낮은 소거활성능을 보였으며, $100\;{\mu}g/mL$의 농도에서 대추과육 추출물은 약 60%, 대추씨 추출물은 약 72%의 소거활성을 보였다. Linoleic acid에 대한 항산화 효과를 측정한 경우에도 FTC가(약 91%)와 TBA가(84-86%)의 값의 차이는 보였으나 추출물의 산화억제 효과가 우수함을 알 수 있었고, 대추과육 및 대추씨 추출물은 DPPH 및 ABTS radical에 대해 높은 소거활성을 보일 뿐만 아니라 hydrogen peroxide의 소거활성능이 뛰어나며 linoleic acid의 산화를 초기단계에서 효과적으로 억제하여 산화의 진행을 상당히 지연시키는 것을 알 수 있었다. V79 cell을 이용하여 $H_2O_2$로 유도된 세포독성에 대한 대추과육 및 대추씨 추출물의 세포보호 효과를 살펴본 결과 $H_2O_2$만 처리한 세포의 생존률이 36% 정도인데 비해 대추과육 및 대추씨 추출물을 함께 처리하였을 때는 생존률이 각각 52, 64%로 증가하여 대추씨 추출물이 $H_2O_2$로 유도된 세포독성에 대해 높은 세포보호 효과를 보였다.

Keywords

References

  1. Wettasinghe M, Shahidi F. Scavenging of reactiveoxigen species and DPPH free radicals by extracts of borage and evening primrose meals. Food Chem. 70: 17-26 (2000) https://doi.org/10.1016/S0308-8146(99)00269-1
  2. Sawyer DT, Valentine JS. How super is superoxide- ACC. Chem. Res. 14: 393 (1981) https://doi.org/10.1021/ar00072a005
  3. Fridorich, I. Biological effects of the superoxide radical. Arch. Biophys. 247: 1-11 (1986) https://doi.org/10.1016/0003-9861(86)90526-6
  4. Ames RN. Dietary carcinogens and anticarcinogens. Oxygen radical and degenerative diseases. Science 221: 1256-1264 (1983) https://doi.org/10.1126/science.6351251
  5. Chance B, Sies H, Boveris A. Hydroperoxide metabolism in mammalian organs. Physiol. Rev. 59: 527-605 (1979) https://doi.org/10.1152/physrev.1979.59.3.527
  6. Kandaswami C, Middleton EJ. Free radical scavenging and antioxidant activity of plant flavonoids. pp. 351-376. In: Free Radicals in Diagnostic Medicine. Armstrong D (ed). Plenum Press, New York and London (1994)
  7. Rhee YK, Kim DH, Han MJ. Inhibitory effect of Zizyphi fructus on ${\beta}$-glucuronidase and tryptophanase of human interestinal bacteria. Korean J. Food Sci. Technol. 30: 199-205 (1998)
  8. Na HS, Kim KS, Lee MY. Effect of jujube methanol extract on the hepatoxicity in CCl4-treated rats. J. Korean Soc. Food Sci. Nutr. 25: 839-845 (1996)
  9. Lee YG, Cho SY. Effect of jujube methanol extract on benzo(a)pyrene induced hepatotoxicity. J. Korean Soc. Food Nutr. 24: 127-132 (1995)
  10. Yook CS. Screening test on the components of the genus Zizyphus in Korea. Korean J. Pharmacog. 3: 27-29 (1972)
  11. Lee SK. Studies on the constituents of the leaves of Zizyphus jujuba Mill. PhD thesis, Pusan National University, Pusan, Korea (1989)
  12. Park MK, Park JH, Shin YG, Cho KH, Han BH, Park MH. Analysis of alkaloids in the seeds of Zizyphus jujuba by high performance liquid chromatography. Arch. Pharm. Res. 14: 99-102 (1991) https://doi.org/10.1007/BF02892010
  13. Folin O, Denis W. On phosphotungastic-phosoho-mdybdic compounds as color reagents. J. Biol. Chem. 12: 239-249 (1912)
  14. Nivea MM, Sampietro AR, Vattuone MA. Comparison of the free radical-scavenging activity of propolis from several regions of Argentina. J. Ethnopharmacol. 71: 109-114 (2000) https://doi.org/10.1016/S0378-8741(99)00189-0
  15. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biol. Med. 26: 1231-1237 (1999) https://doi.org/10.1016/S0891-5849(98)00315-3
  16. Muller HE. Detection of hydrogen peroxide produced by microorganism on ARTS-peroxidase medium. Zentralbl Bakteriol. Mikrobio. Hyg. 259: 151-158 (1985)
  17. Haraguchi H, Hashimoto K, Yagi A. Antioxidative substances in leaves of Polygonum hydropiper. J. Agric. Food Chem. 40: 1349-1351 (1992) https://doi.org/10.1021/jf00020a011
  18. Nakayama T, Niimi T, Osawa T, Kawakishi S. The protective role of polyphenols in cytotoxicity of hydrogen peroxide. Mutat. Res. 281: 77-80 (1992) https://doi.org/10.1016/0165-7992(92)90039-K
  19. Yoshizawa S, Horiuchi T, Yoshida T, Okuda T. Antitumor promoting activity of (-)-epigallocatechin gallate, the main constitutent of tannin in green tea. Phytother. Res. 1: 44-47 (1987) https://doi.org/10.1002/ptr.2650010110
  20. Kim HJ, Jun BS, Kim SK, Cha JY, Cho YS. Polyphenolic compound content and antioxidative activities by extracts from seed, sprout and flower of safflower (Carthamus tinctorius L.). J. Korean Soc. Food Sci. Nutr. 29: 1127-1132 (2000)
  21. Lee YC, Hwang KH, Han DH, Kim SD. Compositions of opuntia ficus-india. Korean J. Food Sci. Technol. 29: 847-853 (1997)
  22. Lee SO, Lee HJ, Yu MH, Im HG. Lee IS. Total polyphenol contents and antioxidant activities of methanol extracts from vegetables produced in Ullung Island. Korean J. Food Sci. Technol. 37: 233-240 (2005)
  23. An BJ, Lee JT, Lee SA, Kwak JH, Park JM, Lee JY, Son JH. Antioxidant effects and application as natural ingredients of Korean sanguisorbae officinalis L. J. Korean Soc. Appl. Biol. Chem. 47: 244-250 (2004)
  24. Lee SE, Seong NS, Bang JK, Park CG, Sung JS, Song J. Antioxidatives activities of Korean medicinal plants. Korean J. Medicinal Crop Sci. 11: 127-134 (2003)
  25. Jung GT, Ju IO, Choi JS, Hong JS. The antioxidative, antimicrobial and nitrite scavenging effects of Schizandra chinensis RUPRECHT (Omija) seed. Korean J. Food Sci. Technol. 32: 928-935 (2000)
  26. Li XF, Li Y, Nam KW, Kim DS, Chio HD, Son, BW. Screening of radical scavenging activity from the marine-derived fungus. Korean J. Pharmacogn. 33: 219-223 (2002)
  27. Rice-Evans CA, Miller NJ. Total antioxidant status in plasma and body fluids. Methods Enzymol. 234: 279-293 (1994) https://doi.org/10.1016/0076-6879(94)34095-1
  28. Miller NJ, Rice-Evans C, Davies MJ, Gopinathan Y, Milner A. A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates. Clin. Sci. 84: 407-412 (1993) https://doi.org/10.1042/cs0840407
  29. Rice-Evans CA, Miller NJ, Paganga G. Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Rad. Biol. Med. 20: 933-956 (1996) https://doi.org/10.1016/0891-5849(95)02227-9
  30. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Rad. Biol. Med. 26: 1231-1237 (1999) https://doi.org/10.1016/S0891-5849(98)00315-3
  31. Ohigashi H, Osawa T, Terao J, Watanabe S, Yoshikawa T. Food Factors for Cancer Prevention. Springer, Tokyo. Japan. pp. 613-616 (1997)
  32. Park SW, Chung SK, Park JC. Active oxygen scavenging activity of Luteolin-7-O-${\beta}$-D-Glucoside Isolated from Humulus japonicus. J. Korean Soc. Food Sci. Nutr. 29: 106-110 (2000)
  33. Farag RS, Badei A, Hewedi FM, Baroty GS. Antioxidant activity of some spice essential oils on linoleic and oxidation in aqueous media. J. Am. Oil Chem. Soc. 66: 792-799 (1989) https://doi.org/10.1007/BF02653670
  34. Mohd ZZ, Abdul-Hamid A, Osman A. Antioxidative activity of extracts from Mengkudu (Morinda citrifolia L.) root, fruit, and leaf. Food Chem. 78: 227-231 (2002) https://doi.org/10.1016/S0308-8146(01)00402-2
  35. Ismail M, Manickam E, Danial AM, Rahmat A, Yahaya A. Chemical composition and anitioxidant activity of Strobilanthes crisp us leaf extract. J. Nutr. Biochem. 11: 536-542 (2000) https://doi.org/10.1016/S0955-2863(00)00108-X
  36. Nakayama T, Hori K, Terazawa K, kawakishi S. Comparision of the cytotoxicity of different hydroperoxides to V79 cells. Free Rad. Res. Commun. 14: 173-178 (1991) https://doi.org/10.3109/10715769109088946
  37. Nakayama T, Yamada M, Osawa T, Kawadishi S. lnhibitory effects of caffeic acid ester in $H_2O_2$ induced cytotoxicity and DNA single strand breaks in Chinese Hamster V79 cells. Biosci. Biotech. Biochem. 60: 316-318 (1996) https://doi.org/10.1271/bbb.60.316
  38. Nakayama T, Yamada M, Osawa T, Kawadishi S. Suppression of active oxygen induced cytotoxicity by flavonoids. Biochem. Pharm. 45: 265-267 (1993) https://doi.org/10.1016/0006-2952(93)90402-I
  39. Branen AS. Toxicology and biochemistry of butylated hydroxtarisole and butylated hydrlxytoluene. J. Am. Oil Chem. Soc. 52: 5963 (1975)