Optimization of Extraction Conditions for Limonin and Nomilin in Citron Seed

유자씨에 함유된 limonin 및 nomilin의 추출 조건 최적화

  • Jeong, Ji-Young (Department of Food Science and Technology, Chungbuk National University) ;
  • Woo, Koan-Sik (National Institute of Crop Science, Rural Development Administration) ;
  • Hwang, In-Guk (Department of Food Science and Technology, Chungbuk National University) ;
  • Lee, Youn-Ri (Department of Food Science and Technology, Chungbuk National University) ;
  • Park, Eui Seok (Metome Food Co. Ltd.) ;
  • Jeong, Heon-Sang (Department of Food Science and Technology, Chungbuk National University)
  • Published : 2008.10.31

Abstract

In this study, response surface methodology was employed in order to optimize the limonin and nomilin extraction conditions from citron seed. The independent variables were extraction temperature, extraction time, and shaking velocity. The yield of limonin and nomilin increased with increased extraction temperature and time. The extraction effects on the limonin and nomilin were higher at extraction temperature (p<0.01). The predicted extraction conditions were validated through actual experiments. The predicted conditions were as follows: $49.7^{\circ}C$ of extraction temperature, 3.3 hr of extraction time, and 400.6 rpm of shaking velocity for the maximum limonin contents (353.9 mg/100 g) and $50.3^{\circ}C$, 3.5 hr, and 399.9 rpm for the maximum nomilin contents (214.5 mg/100 g). The experimental values of limonin and nomilin contents at the predicted conditions were 338.8 and 219.5 mg/100 g, respectively. The predicted values at the optimized conditions were acceptable when compared to the experimental values.

유자씨의 limonin 및 nomilin에 대한 최적 추출조건을 선정하기 위하여 추출온도($30-70^{\circ}C$; $X_1$), 추출시간(1-5 hr; $X_2$) 및 교반속도(200-600 rpm; $X_3$)를 독립변수로 중심합성계획법에 따라 실험설계한 각각의 추출조건에서 limonin 및 nomilin 함량을 측정하였다. Limonin 및 nomilin 추출 조건의 최적화를 위하여 추출용매는 ethanol, 추출용매의 양은 20배 그리고 추출용매 농도는 60%로 선정하였다. Limonin 및 nomilin 추출에 미치는 추출조건의 영향은 추출온도가 가장 컸으며(p<0.01) 그 다음으로는 추출시간으로 나타났다. 반응표면분석결과 최적 추출조건은 limonin에서는 추출온도, 추출시간 및 교반속도가 각각 $49.7^{\circ}C$, 3.3 hr 및 400.6 rpm으로 나타났으며, 이때의 추출량은 353.9 mg/100 g이었다. Nomilin은 각각 $50.3^{\circ}C$, 3.5 hr 및 399.9 rpm이었으며, 추출량은 214.5 mg/100 g이었다. 동일조건에서의 실험값은 limonin 및 nomilin이 가각 338.8 및 219.5 mg/100 g으로 예측값과 실험값이 유사하여 최적화할 수 있었다.

Keywords

References

  1. Nam HW, Hyun YH. Drying of citron juice from by-product of citron tea manufacturing. Korean J. Food Nutr. 16: 334-339 (2003)
  2. Jeong JW, Lee YC, Kim JH, Kim OW. Tnahmgung B. Cooling properties and quality changes during storage of citron (Citrus junos). Korean J. Food Sci. Technol. 28: 1071-1077 (1996)
  3. Bok SH, Lee SH, Park YB, Bae KH, Son KH, Jeong TS, Choi MS. Plasma and hepatic cholesterol and hepatic activities of 3- hydroxyl-3-methyl-glutaryl-CoA reductase and acyl CoA : cholesterol transferase are lower in rats fed citrus peel extract or a mixture of citrus bioflavonoid. Korean J. Food Nutr. 129: 1182-1185 (1999)
  4. Hasegawa S, Miyake M, Ozaki Y. Biochemistry of citrus limonoids and their anticarcinogenic activity. pp. 198-208. In: Food Phytochemicals for Cancer Prevention I. Fruit and Vegetables.Huang MT, Osawa T, Ho CT, Rosen RT (eds.), American Chemical Society Symposium Series 546, Washington DC, USA (1994)
  5. Hasegawa S, Bennett RD, Verdon CP. Limonoids in citrus seeds: origin and relative concentration. J. Agr. Food Chem. 28: 922- 925 (1980) https://doi.org/10.1021/jf60231a016
  6. Ozaki Y, Fong CH, Herman Z, Maeda H, Miyake M, Ifuku Y, Hasegawa S. Limonoid glucosides in citrus seeds. Agr. Biol. Chem. Tokyo 55: 137-141 (1991) https://doi.org/10.1271/bbb1961.55.137
  7. Ohta H, Hasegawa S. Limonoids in pummelos (Citrus grandis (L.) Osbeck). J. Food Sci. 60: 1284-1285 (1995) https://doi.org/10.1111/j.1365-2621.1995.tb04574.x
  8. Vikram A, Jayaprakasha GK, Patil BS. Simultaneous determination of citrus limonoid aglycones and glucosides by high performance liquid chromatography. Anal. Chim. Acta 590: 180-186 (2007) https://doi.org/10.1016/j.aca.2007.03.029
  9. Herman Z, Fong CH, Hasegawa S. Analysis of limonoids in citrus seeds. pp. 361-375. In: Seed Analysis. Modern Methods of Plant Analysis. Linskens HF, Jackson JF (eds). Springer, New York, NY, USA (1992)
  10. Lam LKT, Li Y, Hasegawa S. Effects of citrus limonoids on glutathione- S-transferase activity in mice. J. Agric. Food Chem. 37: 878 (1989) https://doi.org/10.1021/jf00088a010
  11. Kelly C, Jewell C, O'Brien NM. The effect of dietary supplementation with the citrus limonoids, limonin, and nomilin on xenobiotic- metabolizing enzymes in the liver and small intestine of the rat. Nutr. Res. 23: 681-690 (2003) https://doi.org/10.1016/S0271-5317(03)00021-6
  12. Lam LKT, Zhang J, Hasegawa S. Citrus limonoid reduction of chemically induced tumorigenesis. Food Technol. 48: 104-108 (1994)
  13. Kurowska EM, Borradaile N, Meade M, Spence JD, Carroll KK. Cholesterol-lowering effects of dietary citrus juice and their flavonoids. Studies in rats, mice and rabbits. Atherosclerosis 134: 33-37 (1997)
  14. Choi I, Choi S, Nam B, Kim Y, Choi H. Contents of polyphenols and limonoids in citron (Citrus junos Sieb. ex Tanaka) seed extracts and their antioxidant properties. Food Sci. Biotechnol. 17: 373-378 (2008)
  15. Giamperi L, Flaternale D, Bucchini A, Ricci D. Antioxidant activity of Citrus paradise seeds glyceric extract. Fitoterapia 75: 221-224 (2004) https://doi.org/10.1016/j.fitote.2003.12.010
  16. Yoo KM, Lee KW, Park JB, Lee HJ, Hwang IK. Variation in major antioxidants and total antioxidant activity of citron (Citrus junos Sieb ex Tanaka) during maturation and between cultivars. J. Agr. Food Chem. 52: 5907-5913 (2004) https://doi.org/10.1021/jf0498158
  17. Hashinaga F, Herman Z, Hashegawa S. Limonoids in seeds of yuza (Citrus junos Sieb. ex Tanaka). Nippon Shokuhin Kogyo Gakk. 37: 380-382 (1990) https://doi.org/10.3136/nskkk1962.37.5_380
  18. Woo KL, Ha JU, Woo DK. Quantitative determination, separation and purification of limonoid compounds from citron [Citrus junos(L.)]. The Research Institute of Engineering Technology Kyungnam University. Masan, Korea. Theses Collection 15: 363- 372 (1997)
  19. Woo KL, Kim JI, Kim MC, Chang DK. Determination of flavonoid and limonoid compounds in citron (Citrus junos Sieb. ex Tanaka) seeds by HPLC and HPLC/MS. J. Korean Soc. Food Sci. Nutr. 35: 353-358 (2006) https://doi.org/10.3746/jkfn.2006.35.3.353
  20. Park NJ, Kwon JH, Kim HK. Optimization of extraction conditions for ethanol extracts from Chrysanthemum morifolium by response surface methodology. Korean J. Food Sci. Technol. 30: 1189-1196 (1998)