Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Preparative separation of minor saponins from Panax notoginseng leaves using biotransformation, macroporous resins, and preparative high-performance liquid chromatography

  • Liu, Fang (State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau) ;
  • Ma, Ni (Department of Product Development, Wenshan Sanqi Institute of Science and Technology, Wenshan University) ;
  • Xia, Fang-Bo (State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau) ;
  • Li, Peng (State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau) ;
  • He, Chengwei (State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau) ;
  • Wu, Zhenqiang (School of Bioscience and Bioengineering, South China University of Technology) ;
  • Wan, Jian-Bo (State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau)
  • Received : 2017.06.18
  • Accepted : 2017.09.18
  • Published : 2019.01.15
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Abstract

Background: Ginsenosides with less sugar moieties may exhibit the better adsorptive capacity and more pharmacological activities. Methods: An efficient method for the separation of four minor saponins, including gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, from Panax notoginseng leaves (PNL) was established using biotransformation, macroporous resins, and subsequent preparative high-performance liquid chromatography. Results: The dried PNL powder was immersed in the distilled water at $50^{\circ}C$ for 30 min for converting the major saponins, ginsenosides Rb1, Rc, Rb2, and Rb3, to minor saponins, gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, respectively, by the enzymes present in PNL. The adsorption characteristics of these minor saponins on five types of macroporous resins, D-101, DA-201, DM-301, X-5, and S-8, were evaluated and compared. Among them, D-101 was selected due to the best adsorption and desorption properties. Under the optimized conditions, the fraction containing the four target saponins was separated by D-101 resin. Subsequently, the target minor saponins were individually separated and purified by preparative high-performance liquid chromatography with a reversed-phase column. Conclusion: Our study provides a simple and efficient method for the preparation of these four minor saponins from PNL, which will be potential for industrial applications.

Keywords

References

  1. Ma WG, Mizutani M, Malterud KE, Lu SL, Ducrey B, Tahara S. Saponins from the roots of Panax notoginseng. Phytochemistry 1999;52:1133-9. https://doi.org/10.1016/S0031-9422(99)00364-7
  2. Qi LW, Wang CZ, Yuan CS. American ginseng: potential structure-function relationship in cancer chemoprevention. Biochem Pharmacol 2010;80:947-54. https://doi.org/10.1016/j.bcp.2010.06.023
  3. Wan J, Yang F, Li S, Wang Y, Cui X. Chemical characteristics for different parts of Panax notoginseng using pressurized liquid extraction and HPLC-ELSD. J Pharm Biomed Anal 2006;41:1596-601. https://doi.org/10.1016/j.jpba.2006.01.058
  4. Wan JB, Zhang QW, Hong SJ, Li P, Li SP, Wang YT. Chemical investigation of saponins in different parts of Panax notoginseng by pressurized liquid extraction and liquid chromatography-electrospray ionization-tandem mass spectrometry. Molecules 2012;17:5836-53. https://doi.org/10.3390/molecules17055836
  5. Xu YX, Shi JS, Jiang ZL. Inhibitory influence of ginsenoside Rb3 on activation of strychnine-sensitive glycine receptors in hippocampal neurons of rat. Brain Res 2005;1037:99-106. https://doi.org/10.1016/j.brainres.2004.12.044
  6. Xiang H, Liu Y, Zhang B, Huang J, Li Y, Yang B, Huang Z, Xiang F, Zhang H. The antidepressant effects and mechanism of action of total saponins from the caudexes and leaves of Panax notoginseng in animal models of depression. Phytomedicine 2011;18:731-8. https://doi.org/10.1016/j.phymed.2010.11.014
  7. Wang M, Zhang XJ, Liu F, Hu YJ, He CW, Li P, Su HX, Wan JB. Saponins isolated from the leaves of Panax notoginseng protect against alcoholic liver injury via inhibiting ethanol-induced oxidative stress and gut-derived endotoxinmediated inflammation. J Funct Foods 2015;19:214-24. https://doi.org/10.1016/j.jff.2015.09.029
  8. Mao Q, Yi L, L SL, Ye J, Zhang PH, Wang Q. Chemical profiles and anticancer effects of saponin fractions of different polarity from the leaves of Panax notoginseng. Chin J Nat Med 2014;12:30-7. https://doi.org/10.1016/S1875-5364(14)60006-6
  9. Mao Q, Yang J, Cui XM, Li JJ, Qi YT, Zhang PH, Wang Q. Target separation of a new anti-tumor saponin and metabolic profiling of leaves of Panax notoginseng by liquid chromatography with eletrospray ionization quadrupole time-of-flight mass spectrometry. J Pharma Biomed Anal 2012;59:67-77. https://doi.org/10.1016/j.jpba.2011.10.004
  10. Liu F, Ma N, He C, Hu Y, Li P, Chen M, Su H, Wan JB. Qualitative and quantitative analysis of the saponins in Panax notoginseng leaves using ultraperformance liquid chromatography coupled with time-of-flight tandem mass spectrometry and high performance liquid chromatography coupled with UV detector. J Ginseng Res 2018;42:149-57. https://doi.org/10.1016/j.jgr.2017.01.007
  11. Choi SH, Shin TJ, Hwang SH, Lee BH, Kang JY, Kim HJ, Oh JW, Bae CS, Lee SH, Nah SY. Differential effects of ginsenoside metabolites on HERG K+ channel currents. J Ginseng Res 2011;35:191-9. https://doi.org/10.5142/jgr.2011.35.2.191
  12. Kim M, Ahn BY, Lee JS, Chung SS, Lim S, Park SG, Jung HS, Lee HK, Park KS. The ginsenoside Rg3 has a stimulatory effect on insulin signaling in L6 myotubes. Biochem Biophys Res Commun 2009;389:70-3. https://doi.org/10.1016/j.bbrc.2009.08.088
  13. Quan K, Liu Q, Wan JY, Zhao YJ, Guo RZ, Alolga RN, Li P, Qi LW. Rapid preparation of rare ginsenosides by acid transformation and their structureactivity relationships against cancer cells. Sci Rep 2015;5:8598. https://doi.org/10.1038/srep08598
  14. Yang Z, Zhao T, Liu H, Zhang L. Ginsenoside Rh2 inhibits hepatocellular carcinoma through beta-catenin and autophagy. Sci Rep 2016;6, 19383. https://doi.org/10.1038/srep19383
  15. Liu CZ, Yu KH, Sun C, Zhang T, Xu L, Jin Y, Im WT, Jin F. Preparation of minor ginsenosides C-Mx and C-K from notoginseng leaf ginsenosides by a special ginsenosidase type-I. Process Biochem 2015;50:2158-67. https://doi.org/10.1016/j.procbio.2015.10.011
  16. Liu CY, Zhou RX, Sun CK, Jin YH, Yu HS, Zhang TY, Xu LQ, Jin FX. Preparation of minor ginsenosides C-Mc, C-Y, F2, and C-K from American ginseng PPDginsenoside using special ginsenosidase type-I from Aspergillus niger g.848. J Ginseng Res 2015;39:221-9. https://doi.org/10.1016/j.jgr.2014.12.003
  17. Wang L, Liu QM, Sung BH, An DS, Lee HG, Kim SG, Kim SC, Lee ST, Im WT. Bioconversion of ginsenosides Rb(1), Rb(2), Rc and Rd by novel betaglucosidase hydrolyzing outer 3-O glycoside from Sphingomonas sp. 2F2: cloning, expression, and enzyme characterization. J Biotechnol 2011;156:125-33. https://doi.org/10.1016/j.jbiotec.2011.07.024
  18. Han Y, Sun B, Jiang B, Hu X, Spranger MI, Zhang Y, Zhao Y. Microbial transformation of ginsenosides Rb1, Rb3 and Rc by Fusarium sacchari. J Appl Microbiol 2010;109:792-8. https://doi.org/10.1111/j.1365-2672.2010.04707.x
  19. Upadhyaya J, Yoon MS, Kim MJ, Ryu NS, Song YE, Kim YH, Kim MK. Purification and characterization of a novel ginsenoside Rc-hydrolyzing betaglucosidase from Armillaria mellea mycelia. AMB Express 2016;6:112. https://doi.org/10.1186/s13568-016-0277-x
  20. Pang Z, Gao YL, Wang Y, Huang SF. HPLC-UV analysis of main saponins from Panax notoginseng leaves after absorption of 5 macroporous resins. Zhong Cheng Yao 2009;31:1130-2.
  21. Zhang C, Yu H, Bao Y, An L, Jin F. Purification and characterization of ginsenoside-beta-glucosidase from ginseng. Chem Pharm Bull 2001;49:795-8. https://doi.org/10.1248/cpb.49.795
  22. Sunwoo HH, Gujral N, Huebl AC, Kim CT. Application of high hydrostatic pressure and enzymatic hydrolysis for the extraction of ginsenosides from fresh ginseng root (Panax ginseng C.A. Myer). Food Bioprocess Tech 2014;7:1246-54. https://doi.org/10.1007/s11947-013-1234-1
  23. Li H, Liu Y, Yi Y, Miao Q, Liu S, Zhao F, Cong W, Wang C, Xia C. Purification of quercetin-3-O-sophoroside and isoquercitrin from Poacynum hendersonii leaves using macroporous resins followed by Sephadex LH-20 column chromatography. J Chromatogr B 2017;1048:56-63. https://doi.org/10.1016/j.jchromb.2017.01.041
  24. Bai CL, Zhao GR. Separation of salvianic acid A from the fermentation broth of engineered Escherichia coli using macroporous resins. J Sep Sci 2015;38:2833-40. https://doi.org/10.1002/jssc.201500416
  25. Wan JB, Zhang QW, Ye WC, Wang YT. Quantification and separation of protopanaxatriol and protopanaxadiol type saponins from Panax notoginseng with macroporous resins. Sep Purif Technol 2008;60:198-205. https://doi.org/10.1016/j.seppur.2007.08.007

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