Antioxidative Activity and Component Analysis of Fermented Melissa officinalis Extracts

레몬밤 발효추출물의 항산화 활성과 성분 분석

  • Yang, Hee-Jung (Department of Fine Chemistry, College of Nature and Life Science, Seoul National University of Technology) ;
  • Kim, Eun-Hee (Department of Fine Chemistry, College of Nature and Life Science, Seoul National University of Technology) ;
  • Park, Jung-Ok (Department of Fine Chemistry, College of Nature and Life Science, Seoul National University of Technology) ;
  • Kim, Jung-Eun (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) ;
  • 김은희 (서울산업대학교 자연생명과학대학 정밀화학과) ;
  • 박정옥 (서울산업대학교 자연생명과학대학 정밀화학과) ;
  • 김정은 (서울산업대학교 자연생명과학대학 정밀화학과) ;
  • 박수남 (서울산업대학교 자연생명과학대학 정밀화학과)
  • Published : 2009.03.30

Abstract

In this study, the antioxidative effects, inhibitory effects on tyrosinase, and component analysis of fermented Melissa officinalis extracts were investigated. The ethyl acetate fraction of fermented extract ($8.38{\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of M. officinalis. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some M. officinalis extracts on ROS generated in $Fe^{3+}$-EDTA/$H_{2}O_{2}$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of fermented extract ($0.63{\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of M. officinalis on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The M. officinalis extracts suppressed photohemolysis in a concentration dependent manner ($5\;{\sim}\;75{\mu}g/mL$). The inhibitory effect of M. officinalis extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase of some M. officinalis extracts was 50 % ethanol extract ($365{\mu}g/mL$) < ethyl acetate fraction of fermented extract ($122.43{\mu}g/mL$) < ethylacetate fraction ($94.8{\mu}g/mL$). Fractions of ethyl acetate both from ordinary and fermented M. officinalis extracts showed 2 band in TLC and 2 peak in HPLC (330 nm). In HPLC chromatogram of ethyl acetate fraction, peak 1 (51.64 %) and peak 2 (48.36 %) were identified as caffeic acid and rosmarinic acid in the order of elution time. Also, in HPLC chromatogram of ethyl acetate fraction of fermented extract, peak 1 (4.13 %) and peak 2 (95.87 %) were identified as caffeic acid and rosmarinic acid in the order of elution time. These results indicate that the component and content of ordinary and fermented extracts of M. officinalis are different. And the extract of M. officinalis can be used as an antioxidant.

본 연구에서는 레몬밤추출물과 발효추출물의 항산화, 성분 분석 및 tyrosinase 저해 효과에 관한 조사를 수행하였다. 추출물의 free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) 소거활성($FSC_{50}$)은 발효추출물의 ethyl acetate 분획 ($8.38{\mu}g/mL$)에서 가장 큰 활성을 나타내었고, 또한 Luminol-의존성 화학발광법을 이용한 $Fe^{3+}$-EDTA/$H_{2}O_{2}$ 계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 레몬밤추출물의 총 항산화능은 발효추출물의 ethyl acetate 분획($0.63{\mu}g/mL$)에서 가장 큰 활성을 나타내었다. 레몬밤추출물과 발효추출물에 대하여 rose-bengal로 증감된 사람 적혈구의 광용혈에 대한 세포보호 효과를 측정하였다. 레몬밤추출물과 발효추출물의 경우 $5\;{\sim}\;75{\mu}g/mL$의 농도에서 농도 의존적으로 세포보호 효과를 나타내었다. Tyrosinase의 활성 저해 효과($IC_{50}$)는 50 % ethanol 추출물($365{\mu}g/mL$) < 발효추출물의 ethyl acetate 분획($122.43{\mu}g/mL$) < ethyl acetate 분획($94.8{\mu}g/mL$) 순으로 나타났다. 레몬밤추출물 중 ethyl acetate 분획과 발효추출물의 ethyl acetate 분획은 TLC에서 공통으로 2개의 띠로 분리되었다. 또한 HPLC (330 nm)에서도 ethyl acetate 분획과 발효추출물의 ethyl acetate 분획이 각각 2개 peak로 나타났다. 각각의 크로마토그래피로부터 ethyl acetate 분획의 peak 1 (51.64 %)은 caffeic acid, peak 2 (48.36 %)는 rosmarinic acid 임을 확인하였으며, 또한 발효추출물의 ethyl acetate 분획에서도 peak 1 (4.13 %)과 peak 2 (95.87 %)는 각각 caffeic acid와 rosmarinic acid로 확인되었다. 이상의 결과들은 레몬밤의 50 % ethanol 추출물 및 발효추출들이 $^{1}O_{2}$ 혹은 다른 ROS를 소광시키거나 소거함으로써 그리고 ROS에 대항하여 세포막을 보호함으로써 생체계, 특히 태양 자외선에 노출된 피부에서 항산화제로서 작용할 수 있음을 시사한다. 그리고 또한, 레몬밤 추출물과 발효추출물의 성분 및 함량을 분석함으로써 발효 후에 차이점을 확인, 이를 통한 응용 가능성을 확인하였다.

Keywords

References

  1. E. H. Witt, P. Motchnik, and L. Packer, Evidence for UV light as an oxidative stressor in skin, eds J. Fuchs and L. Packer, Oxidative Stress in Dermatology. 29, New York, Dekker (1993)
  2. I. Emerit, Free radicals and aging of the skin, eds L. Emerit and B. Chance, Free radicals and aging, 328, Basel, Birkhauser (1992)
  3. C. S. Foote, Photosensitized oxidation and singlet oxygen; consequences in biological systems, ed. W. A. Pryor, 2, 85, Acdemic press, New York (1976)
  4. S. N. Park, Ph. D. Dissertation, Seoul National Univ., Seoul, Korea (1989)
  5. S. N. Park, Skin aging and antioxidant, J. Soc. Cosmet. Scientists Korea, 23, 75 (1997)
  6. S. N. Park, Protective effect of isoflavone, genistein from soybean on singlet oxygen induced photohemolysis of human erythrocytes, Korean J. Food Sci. Technol., 35(3), 510 (2003)
  7. S. N. Park, Antioxidative properties of baicalein, component from Scutellaria baicalensis Georgi and its application to cosmetics (I), J. Korean Ind. Eng. Chem., 14(5), 657 (2003)
  8. K. Scharffetter-Kochanek, Photoaging of the connective tissue of skin: its prevention and therapy, antioxidants in disease mechanism and therapy, ed. H. Sies, 38, 639 (1997)
  9. R. M. Tyrrell and M. Pidoux, Singlet oxygen involvement in the inactivation of cultured human fibroblast by UVA and near visible radiations, Photochem. Photobiol., 49, 407 (1989) https://doi.org/10.1111/j.1751-1097.1989.tb09187.x
  10. G. F. Vile and R. M. Tyrrell, UVA radiation- induced oxidative damage to lipid and protein in vitro and in human skin fibroblasts is dependent on iron and singlet oxygen, Free Radical Biology & Medicine, 18, 721 (1995) https://doi.org/10.1016/0891-5849(94)00192-M
  11. K. Scharffetter-Kochanek, M. Wlaschek, K. Briviba, and H. Sies, Singlet oxygen induces collagenase expression in human skin fibroblasts, FEBS Lett., 331, 304 (1993) https://doi.org/10.1016/0014-5793(93)80357-Z
  12. M. Wlaschek, K. Briviba, G. P. Stricklin, H. Sies, and K. Scharffetter-Kochanek, Singlet oxygen may mediate the ultraviolet A in induced synthesis of interstitial collagenase, J. Invest. Dermatol., 104, 194 (1995) https://doi.org/10.1111/1523-1747.ep12612751
  13. A. Oikarinen, J. Karvonen, J. Uitto, and M. Hannuksela, Connective tissue alterations in skin exposed to natural and therapeutic UV-radiation, Photodermatology, 2, 15 (1985)
  14. A. Oikarinen and M. Kallioinen, A biochemical and immunohistochemical study of collagen in sun-exposed and protected skin, Photodermatology, 6, 24 (1989) https://doi.org/10.1111/j.1525-1470.1989.tb00262.x
  15. L. H. Kligman, UVA induced biochemical changes in hairless mouse skin collagen: a contrast to UVB effects, ed. F. Urbach, 209, Valdemar, Overland Park (1992)
  16. J. W. Choi, S. I. Kim, J. Y. Kim, H. J. Yang, K. H. Lee, and S. N. Park, Antioxidative and cellular protective effects of Jeju native plant extracts against reactive oxygen species (I), J. Soc. Cosmet. Scientists Korea, 32(3), 181 (2006)
  17. J. W. Choi, S. I. Kim, S. M. Jeon, J. Y. Kim, H. J. Yang, K. H. Lee, and S. N. Park, Antioxidative and cellular protective effects of Jeju plant extracts against reactive oxygen species (Ⅰ), J. Soc. Cosmet. Scientists Korea, 32(3), 181 (2006)
  18. H. J. Yang and S. N. Park, Evaluation of antioxidant potential of extract/fractions of Equisetum arense (Ⅰ), J. Soc. Cosmet. Scientists Korea, 33(2), 61 (2007)
  19. H. J. Yang and S. N. Park, Component analysis and study on anti-elastase activity of Equisetum arense (Ⅱ), J. Soc. Cosmet. Scientists Korea, 33(3), 139 (2007)
  20. S. M. Jeon, S. I. Kim, J. Y. Ahn, and S. N. Park, Antioxidtive potenties of extract/fractions of Suaeda asparagoides and Salicornia herbacea extracts (Ⅰ), J. Soc. Cosmet. Scientists Korea, 33(3), 145 (2007)
  21. J. Y. Kim, H. J. Yang, K. H. Le, S. M. Jeon, Y. J. Ahn, B. R Won, and S. N. Park, Antioxidative and antiagiang effect of Jeju native plant extracts (Ⅱ), J. Soc. Cosmet. Scientists Korea, 32(3), 181 (2006)
  22. Anon, Iranian herbal pharmacopoeia, Tehran: Ministry of Health Publication, 1, 141 (2002)
  23. N. D. Allahverdiyeva, M. Ozguvenc, and S. Koltasd, Antiviral activity of the volatile oils of Melissa officinalis L. against Herpes simplex virus type-2, Phytomedicine, 11, 657 (2004) https://doi.org/10.1016/j.phymed.2003.07.014
  24. M. D. Neda, B. Biljana, S. Marina, and S. Natasa, Antimicrobial and antioxidant activities of Melissa officinalis L. (Lamiaceae) essential oil, J. Agric. Food Chem., 52, 2485 (2004) https://doi.org/10.1021/jf030698a
  25. K. Dastmalchi, H. J. Damien Dormana, P. P. Oinonena, Y. Darwisd, I. Laakso, and R. Hiltunena, Chemical composition and in vitro antioxidative activity of a lemon balm (Melissa officinalis L.) extract, K. Dastmalchi et al., LWT 41, 391 (2008) https://doi.org/10.1016/j.lwt.2007.03.007
  26. J. L. Lamaison, C. Petitjean-Freytet, and A. Carnat, Rosmarinic acid, total hydroxycinnamic derivatives and antioxidant acivity of Apiaceae, Borraginaseae and Lamiceae medicinals, Ann. Pharm. Fr., 48, 103 (1990)
  27. J. S. Kim, B. R. Park, E. K. Park, H. S. Lee, J. C. Hahm, K. H. Bae, and M. Y. Kim, Screening of anti-angiogenic activity from plant extracts, Kor. J. Pharmacogn., 37(4), 253 (2006)