Natural Product Sciences

The Korean Society of Pharmacognosy (한국생약학회)
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Development of Gradient Centrifugal Partition Chromatography Method and Its Application for the Isolation of 3,5-Dimethoxyphenanthrene-2,7-diol and Batatasin-I from Dioscorea opposita

  • Yoon, Kee-Dong (College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University) ;
  • Yang, Min-Hye (College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University) ;
  • Chin, Young-Won (College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University) ;
  • Kim, Yoen-Jun (College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University) ;
  • Kim, Hye-Ryung (College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University) ;
  • Choi, Ki-Ri (College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University) ;
  • Park, Ju-Hyun (R&D Institute, Tong Yang Moolsan Co., Ltd.) ;
  • Kim, Jin-Woong (College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University)
  • Published : 2009.09.30
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Abstract

Gradient centrifugal partition chromatography (GCPC) method was developed and applied to isolate 3,5-dimethoxyphenanthrene-2,7-diol (DMP) and batatasin-I (BA-I) from the dichloromethane soluble extract of Dioscorea opposita. In this method, the lower phase of n-hexane-methanol-water system (HMW, 10 : 9 : 1, v/v) was used as a mobile phase A (MpA) and water was used as a mobile phase B (MpB). This gradient CPC method is comparable to that of reversed-phase HPLC method in that the stationary upper-phase of HMW (10 : 9 : 1 v/v) works as if it were reversed-phase silica gel due to its hydrophobic property, while the lower phase (MpA) and water (MpB) functioned as hydrophilic mobile phases. The initial condition of the mobile phase was 20% MpA/80% MpB and maintained for 150 min to obtain DMP (1.2 mg), and then MpA was increased up to 50% to elute BA-I (1.7 mg). The purities of DMP and BA-I were 94.1% and 98.3% with the recovery yields of 83% and 86%, respectively. Similar results were obtained by linear-gradient CPC. The CPC peak fractions were identified by comparing their retention time to those of authentic samples of DMP and BA-I and their spectroscopic data ($^1$H NMR and $^{13}$C NMR) to those of literature values.

Keywords

References

  1. Akahori, A., Studies on the steroidal components of domestic plants- XLIV.: Steroidal sapogenins contained in Japanese Dioscorea sp. Phytochemistry, 4, 97-106 (1965) https://doi.org/10.1016/S0031-9422(00)86151-8
  2. Bae, K., The Medicinal Plants of Korea, Kyo-Hak, Seoul, p. 551 (1999)
  3. Chu, X., Sun, A., and Liu, R., Preparative isolation and purification of five compounds from the Chinese medicinal herb Polygonum cuspidatum Sieb. et Zucc by high-speed counter-current chromatography. J. Chromatogr. A, 1097, 33-39 (2005) https://doi.org/10.1016/j.chroma.2005.08.008
  4. Cortez, D.A.G., Marston, A., and Hostettmann, K., Separation of xanthones and a biphenyl from Kielmeyera coriacea centrifugal partition chromatography. Chromatographia, 50, 7- 10 (1999) https://doi.org/10.1007/BF02493609
  5. Coursey, D.E.G., Yam storage-I: A review of yam storage practices and of information on storage losses. J. Stored. Prod. Res., 2, 229-244 (1967) https://doi.org/10.1016/0022-474X(67)90070-7
  6. Coxon, D.T., Ogundana, S.K., and Dennis, C., Antifungal phenanthrenes in yam tubers. Phytochemistry, 21, 1389-1392 (1982) https://doi.org/10.1016/0031-9422(82)80148-9
  7. Du, Q., Jerz, G., Chen, P., and Winterhalter, P., Preparation of ursane triterpenoids from Centella asiatica using high speed countercurrent chromatography with step-gradient elution. J. Liq. Chromatogr. Relat. Technol., 27, 2201-2215 (2004) https://doi.org/10.1081/JLC-200025707
  8. Foucault, A.P., Theory of Centrifugal Partition Chromatography: Centrifugal Partition Chromatography, Marcel Dekker, New York (1995)
  9. Fu, Y.-C., Ferng, L.-H., and Huang, P.-Y., Quantitative analysis of allantoin and allantoic acid in yam tuber, mucilage, skin and bulbil of the Dioscorea species. Food Chem., 94, 541-549 (2006) https://doi.org/10.1016/j.foodchem.2004.12.006
  10. Fu, Y.-C., Huang, P.-Y., and Chu, C.-J., Use of continuous bubble separation process for separating and recovering starch and mucilage from yam (Dioscorea pseudojaponica yamamoto). LWT-Food Sci. Technol., 38, 735-744 (2005) https://doi.org/10.1016/j.lwt.2004.09.008
  11. Himbert, F., Pennanec, R., Guillaumet, G., and Lafosse, M., Preparative liquid chromatography and centrifugal partition chromatography for purification of new anticancer precursors. Chromatographia, 60, 269-274 (2004) https://doi.org/10.1365/s10337-004-0303-z
  12. Jayakody, L., Hoover, R., Liu, Q., and Donner, E., Studies on tuber starches II. Molecular structure, composition and physicochemical properties of yam (Dioscorea sp.) starches grown in Sri Lanka. Carbohydr. Polym., 69, 148-163 (2007) https://doi.org/10.1016/j.carbpol.2006.09.024
  13. Kim, C.Y. and Kim, J., Preparative isolation and purification of geniposide from gardenia fruits by centrifugal partition chromatography. Phytochem. Anal., 18, 115-117 (2007) https://doi.org/10.1002/pca.958
  14. Leitao, G., El-Adji, S.S., Lopes de Melo, W.A., Leitao, S.G., and Brown, L., Separation of free and glycosylated flavonoids from Siparuna guianensis by gradient and isocratic CCC. J. Liq. Chromatogr. Relat. Technol., 28, 2041- 2051(2005) https://doi.org/10.1081/JLC-200063669
  15. Leong, Y.-W., Kang, C.-C., Harrison, L.J., and Powell, A.D., Phenanthrenes, dihydrophenanthrenes and bibenzyls from the orchid Bulbophyllum vaginatum. Phytochemistry, 44, 157- 165 (1997) https://doi.org/10.1016/S0031-9422(96)00387-1
  16. Liu, A., Li, A., and Sun, J., Preparative isolation and purification of coumarins from Angelica dahurica (Fisch. ex Hoffm) Benth, et Hook. f (Chinese traditional medicinal herb) by high-speed counter-current chromatography. J. Chromatogr. A, 1052, 223- 227 (2004a) https://doi.org/10.1016/j.chroma.2004.08.127
  17. Liu, R., Lei, F., Sun, A., and Kong, L., Preparative isolation and purification of coumarins from Peucedanum praeruptorum Dunn by high-speed counter-current chromatography. J. Chromatogr. A, 1057, 89-94 (2004b) https://doi.org/10.1016/j.chroma.2004.09.047
  18. Marchal, L., Legrand, J., and Foucault, A., Centrifugal partition chromatography: A survey of its history, and our recent advances in the field. Chem. Rec., 3, 133-143 (2003) https://doi.org/10.1002/tcr.10057
  19. Murayama, W., Kobayashi, T., Kosuge, Y., Yano, H., Nunogaki, Y., and Nunogaki, K., A new centrifugal counter-current chromatograph and its applications. J. Chromatogr., 239, 643- 649 (1982) https://doi.org/10.1016/S0021-9673(00)82022-1
  20. Oliveira, R., Leitao, G.G., Moraes, M.C.C., Kaplan, M.A.C., Lopes, D., and Jorge, J.P.P., Gradient elution for triterpene separation from Cecropia lyratiloba Miquel by HSCCC. J. Liq. Chromatogr. Relat. Technol., 28, 1985-1992 (2005) https://doi.org/10.1081/JLC-200063644
  21. Takasugi, M., Kawashima, S., Monde, K., Katusi, N., Masamune, T., and Shrrata, A., Antifungal compounds from Dioscorea batatas inoculated with Pseudomonas cichorii. Phytochemistry, 26, 371-375 (1987) https://doi.org/10.1016/S0031-9422(00)81417-X
  22. Yoon, K.D. and Kim, J., Preparative separation of dioscin derivatives from Dioscorea villosa by centrifugal partition chromatography coupled with evaporative light scattering detection. J. Sep. Sci., 31, 2486-2491 (2008) https://doi.org/10.1002/jssc.200800136
  23. Yoon, K.D., Yang, M.H., Nam, S.I., Park, J.H., Kim, Y.C., and Kim, J., Phenanthrene derivatives, 3,5-dimethoxyphenanthrene-2,7-diol and batatasin-I, as non-polar standard marker compounds for Dioscorea rhizoma. Nat. Prod. Sci., 13, 378-383 (2007)
  24. Zhang, L., Liu, Y., and Chen, G., Simultaneous determination of allantoin, choline and -arginine in Rhizoma Dioscoreae by capillary electrophoresis. J. Chromatogr. A, 1043, 317- (2004) https://doi.org/10.1016/j.chroma.2004.06.003
  25. Zhao, G., Kan, J., Li, Z., and Chen. Z., Structural features and immunological activity of a polysaccharide from Dioscorea opposita Thunb roots. Carbohydr. Polym., 61, 125-131 (2005) https://doi.org/10.1016/j.carbpol.2005.04.020