The Study on the Whitening Effect of Ephedra sinica Extract

마황추출물의 미백효과에 관한 연구

  • 유영근 (한국화장품(주) 기술개발연구소) ;
  • 정민석 (한국화장품(주) 기술개발연구소) ;
  • 최종완 (한국화장품(주) 기술개발연구소) ;
  • 김중회 (한국화장품(주) 기술개발연구소)
  • Published : 2005.06.01

Abstract

In this study, we investigated the application of an extract from Ephedra sinica which has been composed of traditional Korean medicine as a whitening ingredient. The extract of Ephedra sinica which was obtained from the mixture of methanol and water (1:1) the inhibitory effect of tyrosinase. Then, Ephedra sinica was extracted by two different solvents. One was water and the other was methylene chloride. Only, the water extract of Ephedra sinica showed the inhibitory effort of tyrosinase; the anti-tyrosinase activity with $0.2\%$ of the water extract was $60.6\%$. But the extract of Ephedra sinica in methylene chloride fraction showed little inhibitory effect on tyrosinase. The inhibitory effect of the concentrated water extract of Ephedra sinica was tested on L-DOPA auto-oxidation and melanin synthesis in B-16 melanoma. In L-DOPA auto-oxidation, $0.5\%$ of the concentrated water extract showed $87\%$ of inhibition of L-DOPA auto-oxidation and the $0.75\%$ concentrated Ephedra sinica extract in wafer fraction inhibited $98.8\%$ of that. In melanin synthesis of B-16 melanoma, the concentrated water effect of Ephedra sinica inhibited $70.2\%$ or $79.9\%$ of inhibitory effect on that at the concentration of $0.05\%$ or $0.075\%$, respectively. For verifying the skin whitening effect of the concentrated water extract of Ephedra sinica in vivo, we performed the clinical test of that. The research showed the significant skin whitening effect of a cream containing $0.5\%$ Ephedra sinica extract and the statistical analysis showed a significant difference (p < 0.05) between sample (containing $0.5\%$ Ephedra sinica extract) and placebo after 10 weeks.

본 연구는 한방제제의 구성 약재인 마황이라는 천연한방소재로부터 미백효과가 있는 화장품 원료를 개발하고자 하였다. 본 실험에서 마황추출물의 tyrosinase 억제 효과를 확인하고 마황추출 과정을 세분화하여 methylene chloride 및 물분획물을 얻었으며 이들을 가지고 다시 tyrosinase 억제 실험을 실시하였다. 그 결과 수층부분에서만 $0.2\%$ 농도에서 $60.6\%$의 tyrosinase 억제 효과를 보여주었으며 이후 수층부분만을 농축하여 L-DOPA 산화억제 실험 및 B-16 melanoma를 이용한 미백 실험을 실시하였다. 그 결과 마황추출물 $0.5\%$ 농도에서 $87\%$의 L-DOPA 산화억제 효과를 보여주었으며, $0.75\%$에서는 $98.8\%$의 억제효과를 보여주었다. 또한 B-16 melanoma에서는 $0.05\%$에서 $70.2\%$, $0.075\%$에서는 $79.9\%$의 억제효과를 보여주었다. 그리고 수층부분만을 농축한 마황추출물의 in vivo상에서의 미백 효과를 검증하기 위하여 마황추출물 $0.5\%$를 함유한 제형으로 미백 임상실험을 실시하였다. 그 결과 마황추출물을 함유한 제형에서 10주 경과 후에 육안 및 기기평가 모두에서 미백효과를 보여주었으며 통계적으로도 유의한 차이(p<0.05)를 보여주었다.

Keywords

References

  1. Urbach and Frederick, The biologic effects of ultraviolet radiation emphasis on skin, 1st international conference on skin & cancer, hospital temple university health science, Pergamon, N.Y. (1969)
  2. A. Libert, Use of alpha-melanocyte-stimulatinghormone analogue to improve alpha-melanocytestimulating-hormone receptor binding assay in human melanoma, Pigment cell Res., 2, 510 (1989) https://doi.org/10.1111/j.1600-0749.1989.tb00247.x
  3. G. Prota, Melanins, and Melanogenesis, Academic Press, N.Y. (1992)
  4. V. J. Hearing and M. Jimenez, Analysis of pigmentation at the molecular level, Pigment cell Res., 2, 95 (1989)
  5. H. Matsuda, M. Higashino, Y. Nakai, M. Iinuma, M. Kubo, and F. A. Lang, Studies of cuticle drugs from natural sources. IV. inhibitory effects of some Arctostaphylos plants on melanin biosynthesis, Bio. Pharm. Bull., 19(1), 153 (1996) https://doi.org/10.1248/bpb.19.153
  6. T. Tsuruga, Y. T. Chun, Y. Ebizuka, and U. Sankawa, Biologically active constituents of Melaleuca leucadendron: inhibitors of induced histamine release from rat mast cells, Chem Pharm Bull., 39(12), 3276 (1991) https://doi.org/10.1248/cpb.39.3276
  7. F. Qiu, K. I. Komatsu, K. I. Saito, K. Kawasaki, X. Yao, and Y. Kano, Pharmacological properties of traditional medicine. XIII. pharmacokinetic study of mulberroside A and its metabolites in rat, Biol. Pbarm. Bull., 19(11), 1463 (1996) https://doi.org/10.1248/bpb.19.1463
  8. T. Ikeda and T. Tsutsumi, Function and skin depigmental activity of crude drugs, Fragrance J., 6, 59 (1990)
  9. T. Yokota, H. Nishio, Y. Kubota, and M. Mizoguchi, The inhibitory effect of glabridin from licorice extracts on melanogenesis and inflammation, Pigment Cell Research, 11, 355 (1998) https://doi.org/10.1111/j.1600-0749.1998.tb00494.x
  10. A. J. Ramsay, F. Lynn, N. A. Iscoe, and H. J. Kahn, MIB-1 proliferative activity is a significant prognostic factor in primary thick cutaneous melanomas, J. Invest. Dermatol., 105(1), 22 (1995) https://doi.org/10.1111/1523-1747.ep12312431
  11. Y. Yada, K. Higuchi, and G. Imokawa, Effects of endothelins on signal transduction and proliferation in human melanocytes, J. Biol Chem., 266(27), 18352 (1991)
  12. J. R. Gruber, S. Ohno, and R. M. Niles, Increased expression of protein kinase C alpha plays a key role in retinoic acid-induced melanoma differentiation, J. Biol. Chem., 267(19), 13356 (1992)
  13. K, Maeda and M. Fkuda, In vitro effectiveness of several whitening cosmetic components in human melanocyte, J. Soc. Cosmet. Chem., 42, 361 (1991)
  14. S. Pavel, Dynamics of melanogenesis intermediates, J. Invest. Dermatol., 100(2), 1523 (1993)
  15. K, Maeda and M. Fkuda, Arbutin: mechanism of its depigmentation in human melanocyte culture, American Soc. Pharm. Exp. Therapeutics, 276(2), 765 (1996)
  16. K, Maeda and M. Fkuda, In vitro effectiveness of several whitening cosmetic components in human melanocyte. J. Soc. Cosmet. Chem., 42, 361 (1991)
  17. Y. Masamoto, S. Iida, and M. Kubo, Inhibitory effect of Chinese crude drugs on tyrosinase, Planta. Med., 40(4), 361 (1980) https://doi.org/10.1055/s-2008-1074986
  18. Y. Ishihara, M. Oka, M. Tsunakawa, K. Tomita, M. Hatori, H. Yamamoto, H. Kamei, T. Miyaki, Koni, and T. Oki, Melanostatin, a new melanin synthesis inhibitor. Production, isolation, chemical properties, structure and biological activity, J. Antibiotics, 44, 25 (1991) https://doi.org/10.7164/antibiotics.44.25
  19. H. S. Mason and E. W. Peterson, Melanoproteins reactions between enzyme-generated quinones and amino acids. Biochem Biophys. Acta, 111, 134 (1965) https://doi.org/10.1016/0304-4165(65)90479-4
  20. P. R. Gorden, C. P. Mansur, and B. A. Gilchrest, Regulation of human melanocyte growth, dendricity and melanization by keratinocyte derived factors, J. Invest. Dermatol., 92, 565 (1989) https://doi.org/10.1111/1523-1747.ep12709595
  21. T. Mosmann, Rapid colorimetric assay for cellular growth and survival, J. Immunol. Methods, 65, 55 (1983) https://doi.org/10.1016/0022-1759(83)90303-4
  22. A. Garcia and J. E. Fulton, The combination of glycolic acid and hydroquinone or kojic acid for the treatment of melanoma and related conditions, Dermatol. Surg., 22(5), 443 (1996)
  23. C. Cotellessa, K. Peris, M. T. Onorati, M. C. Fargnoli, and S. Chimenti, The use of chemical peelings in the treatment of different cutaneous hyperpigmentations, Dermatol. Surg., 25(6), 450 (1999) https://doi.org/10.1046/j.1524-4725.1999.08217.x
  24. 김상태, 서기석, 채영수, 엄상철, Arbutin, glycolic acid, kojic acid 및 pentadecenoic acid가 in vitro 및 in vitro 에서 UVB 조사에 의한 색소형성에 미치는 영향. 대한 피부과학회지, 32(6), 29 (1994)
  25. J. Serup and T, Agner, Colorimetric quantification of erythema: a comparison of two colorimeters with a clinical scoring scheme and laser-dopper flowmetry, Clin. Exp. Dermatol., 15, 267 (1990) https://doi.org/10.1111/j.1365-2230.1990.tb02087.x
  26. V. J. Hearing and K. Jsukamoto, Enzymatic control of pigmentation in mammals, FASEB J., 5, 2902 (1991)