Optimization of Extraction Conditions to Obtain Functional Components from Buckwheat (Fagopyrum esculentum M.) Sprouts, using Response Surface Methodology

반응표면분석법에 의한 메밀(Fagopyrum esculentum M.) 새싹 기능성분의 추출 조건 최적화

  • Published : 2009.10.30

Abstract

Response surface methodology (RSM) was used to optimize extraction conditions for functional components of buckwheat (Fagopyrum esculentum). A central composite design was applied to investigate the effects of three independent variables, namelyextraction temperature (X1), extraction time (X2), and ethanol concentration (X3), on responses including extraction yield (Y1), total phenolic content in the extract (Y2), $\alpha$-glucosidase inhibition activity (Y3), and acetylcholine esterase (ACE) inhibition activity (Y4). Data were analyzed using an expert design strategy and statistical software. The maximum yield was 24.95% (w/w) at $55.75^{\circ}C$ extraction temperature, 8.75 hextraction time, and 15.65% (v/v) ethanol. The maximum total phenolic yield was 222.45 mg/100 g under the conditions of $28.11^{\circ}C$ extraction temperature, 8.65 h extraction time, and 81.72% (v/v) ethanol. The maximum $\alpha$-glucosidase inhibition activity was 85.38% at $9.62^{\circ}C$, 7.86 h, and 57.58% (v/v) ethanol. The maximum ACE inhibition activity was 86.91% under extraction conditions of $10.12^{\circ}C$, 4.86 h, and 44.44% (v/v) ethanol. Based on superimposition of a four-dimensional RSM with respect to levels of total phenolics, $\alpha$-glucosidase inhibition activity, and ACE inhibition activity, obtained under various extraction conditions, the optimum ranges of conditions were an extraction temperature of $0-70^{\circ}C$, an extraction time of 2-8 h, and an ethanol concentration of 30-80% (v/v).

본 실험은 메밀새싹의 추출조건에 따른 수율, 총 페놀, $\alpha$-glucosidase 저해활성능, 및 ACE 저해활성능에 대해 반응표면 분석법을 이용하여 추출조건을 최적화하였다. 중심합성계획에 따라 추출온도($0{\sim}100^{\circ}C$), 추출시간(0~12 hr) 및 에탄올 농도(0~100%)를 달리하였을 때 반응 표면 회귀식의 R2는 수율, 총 페놀, $\alpha$-glucosidase 저해활성능 및 ACE 저해활성능에서 각각 0.9461(p<0.001), 0.8875(p<0.005), 0.9186(p<0.005) 및 0.8667(p<0.005)로 나타내었으며, 추출조건별 총 페놀 함량, $\alpha$-glucosidase 저해활성능 및 ACE 저해활성능에 대한 반응표면을 superimposing하여 얻은 최적 추출조건 범위는 추출온도 $0{\sim}70^{\circ}C$, 추출시간 2~8 hr 및 에탄올 농도 30~80%로 나타내었다. 최적 추출조건 범위내의 임의의 조건인 추출온도 $15^{\circ}C$, 추출시간 5 hr 및 에탄올 농도 50 %를 회귀식에 대입하여 얻은 예측값은 수율 15.18%, 총 페놀 함량 189.27 mg/100 g, $\alpha$-glucosidase 저해활성능 77.44 % 및 ACE 저해활성능 85.24%으로 예측되었으며, 실제 실험을 통해 얻어진 값은 수율 16.18%, 총 페놀 함량 175.57 mg/100 g, $\alpha$-glucosidase 저해활성능 79.17% 및 ACE 저해활성능 81.60%으로 매우 유사하게 나타났다.

Keywords

References

  1. Hao, X., Li, G., Yang, W., Zhou, N., Nin, R. and Zhou, M. (1995) The difference and clasification of light reaction of buckwheat under different treatments of light duration - First report of light reaction difference of buckwheat varieties. International symposium on buckwheat 1995, Current Advances in Buckwheat Research, 541-549
  2. Wang, Y., Liu, J. and Qingin, D. (1995) The study on nitrogen and phosphorus content in buckwheat plant and fertilizer application index. International symposium on buckwheat 1995, Current Advances in Buckwheat Research, 593-599
  3. Havsteen, B. (1983) Flavonoids a class of natural products of high pharmacological potency. Biochem. Pham., 32, 1141-1148 https://doi.org/10.1016/0006-2952(83)90262-9
  4. Lee. J.S., Maeng. Y.S. and Ju, J.S. (1992) The effect of buckwheat supplement on metabolic status of streptozotocin induced diabetic rats, Annual Report of Korea Nutr. Hallym Univ., 9, 21-25
  5. Marshall, H.G. and Pomeranz, Y. (1982) Buckwheat description, breeding, production and utilization. In Y. Pomeranz, ed., Advances in Cereal Science and Technology. Am. Assoc. Cereal Chem., St. Paul, MN. USA, pp 157-212
  6. Choi, Y.S., Ahn, A., Shim, H.H., Choe, M. and Oh, S.Y. (1992) Effects of instant buckwheat noodle on digestability and lipids profiles liver and serum in rats. J. Korean Soc. Food Nutr., 21, 478-483
  7. Ikeda, K. (1997) Buckwheat: Utilization and recent progress in research. AACC Pacific Rim News, 4-12
  8. Mazza, G. and Campbell, C.G. (1985) Influence of water activity and temperarture on dehulling of buckwheat. Cereal Chem., 62, 31-35
  9. Mazza, G. (1988) Lipid content and fatty acid composition of buckwheat seed. Cereal Chem., 65, 122-126
  10. Lee, E.H. (2007) Changes in physico-chemical characteristics during buckwheat germination. Master's degree thesis. Sookmyung Women's University
  11. Kim, Y.S., Kim, J.G., Lee, Y.S. and Kang, I.J. (2005) Comparison of the chemical components of buckwheat seed and sprout. J. Korean Soc. Food Sci. Nutr., 34, 81-86 https://doi.org/10.3746/jkfn.2005.34.1.081
  12. Kim, S.L., Son, Y.K., Hwang, J.J., Kim, S.K. and Hur, H.S. (1998) Development of buckwheat sprout as a functional vegetable. RDA. J. Crop Sci., 40, 191-199
  13. SAS Institute, Inc. (1998) SAS/STAT User's Guide, Statistical Analysis Systems Institute, Cary, NC, USA
  14. Martha, L.A. and James, P.B. (1992) The Mathematica handbook, Compatible with Mathematica, Version 2.0 Harcourt brace, Massachusetts : An Imprint of a Division of Academic Press
  15. AOAC (1995) Official Methods of Analysis. 16th ed. Method 952.03 Association of Official Analytical Communities, Arlington, VA, USA. Chapter 26, pp 16-17
  16. Ogawa, S., Fujieda, S., Sakata, Y., Ishizaki, M., Hisamatsu, S., Okazaki, K., Ooki, Y., Mori, M., Itoh, M. and Korenaga, T. (2004) Synthesis and glycosidase inhibitory activity of some N-substituted 5a-carba-β -fuco- and β-galactopyranosyl-amines, and selected derivatives. Bioorg. Med. Chem., 12, 6569-6579 https://doi.org/10.1016/j.bmc.2004.09.023
  17. Cushman, D.W. and Cheung, H.S. (1971) Spectrophotometric assay and properties of the angiotensin converting enzyme of rabbit lung. Biochem. Pharmacol., 20, 1637-1648 https://doi.org/10.1016/0006-2952(71)90292-9
  18. Kim, N.M., Ko, S.R., Choi, K.J. and Kim, W.J. (1993) Effect of some factors on extraction of effectual components in cinnamon extracts. J. Korean Agric. Chem. Soc., 36, 17-22
  19. Park, N.Y., Lee, G.D., Jeong, Y.J. and Kwon, J.H. (1998) Optimization of extraction conditions for physicochemical properties of ethanol extracts from Chrysanthemum boreale. J. Korean Soc. Food Sci. Nutr., 27, 585-590
  20. Lee, T.B. (1979) Illustrated Folra of Korea. Hyangmoon Publishing Co., Seoul, Korea. pp 511
  21. Park, J.W., Kim, H.S., Park, I.B., Shin, G.W., Lee, Y.J. and Jo, Y.C. (2007) Optimization of ethanol extraction conditions from glasswort(Salicornia herbacea) using response surface methodology. Korean J. Food Preserv., 16, 376-384
  22. Lee, G.D., Kim, J.O., Joo, G.J. and Kwon, J.H. (2005) Optimum conditions for the extraction of effective substances from the stem of Opuntia fisccus-indica. Food Sci. Biotechnol., 14, 190-195
  23. Josse, R.G., Chiasson, J.L., Ryan, E.A., Lau, D.C., Ross,S.A., Yale, J.F., Leiter, L.A., Maheux, P., Tessier, D., Wolever, T.M., Gerstein, H., Rodger, N.W., Dornan, J.M., Murphy, L.J., Rabasa-Lhoret, R. and Meneilly, G.S. (2003) Acarbose in the treatment of elderly patients with type 2 diabetes. Diabetes Res. Clin. Pract., 59, 37-42 https://doi.org/10.1016/S0168-8227(02)00176-6
  24. Chung, W.T., Lee, S.H., Cha, M.S., Sung, N.S., Hwang, B. and Lee, H.Y. (2001) Biological activites in roots of Glycyrrhiza uralensis Fisch. Korean J. Med. Crop Sci., 9, 45-54
  25. Kim, J.I., Kang, M.J. and Bae, S.Y. (2003) Hypoglycemic effect of the methanol extract of soybean sprout in streptozotocin-induced diabetic rats. J. Korean Soc. Food Sci. Nutr., 32. 921-925 https://doi.org/10.3746/jkfn.2003.32.6.921
  26. Oh, S.J., Kim, S.H., Kim, S.K., Baek, Y.J. and Cho, K.H. (1997) Angiotensin Ⅰ-converting enzyme inhibitory activity of the K-casein fragments hydrolyzated by chymosin, pepsin, and trysin. Fractionation of angiotensin converting enzyme(AEC) inhigitory peptides from soybean paste. Korean J. Food Sci. Technol., 27, 230-234
  27. Kayashita, J., Shimaoka, I., Nakajoh, M., Kishida, N. and Kato, N. (1999) Consumption of a buckwheat protein extract retards 7, 12-dimethylbenz(α)anthracene-induced mammary carcenogenesis in rats. Biosci. Bitechnol. Biochem., 63, 1837-1839 https://doi.org/10.1271/bbb.63.1837
  28. Lee, M.H., Yoon, S.R., Jo, D.K., Kim, H.K. and Lee, G.D. (2007) Optimization of extraction conditions for functional components of roasted Pleurotus eryngii by microwave-assisted extraction. J. Korean Soc. Food Sci. Nutr., 36, 1062-1069 https://doi.org/10.3746/jkfn.2007.36.8.1062
  29. Nakamura, K., Maejima,, Y. and Maejima, S. (2006) Isolation of hydrophilic ACE inhibitory peptides from fermented buckwheat sprout. Peptide Sci., 42, 191-194