Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Identification and quantification of major malonyl ginsenosides isolated from Panax ginseng C.A. Meyer

인삼(Panax ginseng C.A. Meyer)로부터 Malonyl ginsenoside의 분리 및 정량분석

  • Shin, Woo Cheol (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Jung, Jiyun (Punggi Ginseng Research Institute, GBARES) ;
  • Na, Hyeon Seon (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Bo, Jeon Hwang (Graduate School of Biotechnology and Department of Oriental Medicine Biotechnology, Kyung Hee University) ;
  • Kim, Hyoung-Geun (Graduate School of Biotechnology and Department of Oriental Medicine Biotechnology, Kyung Hee University) ;
  • Yoon, Dahye (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Choi, Bo-Ram (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Lee, Young-Seob (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Kim, Geum-Soog (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Baek, Nam-In (Graduate School of Biotechnology and Department of Oriental Medicine Biotechnology, Kyung Hee University) ;
  • Lee, Yi (Department of Industrial Plant Science and Technology, Chungbuk National University) ;
  • Lee, Dae Young (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA)
  • Received : 2019.10.14
  • Accepted : 2019.10.25
  • Published : 2019.12.31
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Abstract

The root of Panax ginseng C.A. Meyer were extracted with 70% aqueous EtOH and the concentrates were partitioned into MeOH and H2O fractions using Diaion HP-20. The repeated SiO2 or octadecyl SiO2 column, and MPLC for the MeOH fraction led to isolation of four malonyl ginsenosides. The chemical structures of these compounds were determined as malonyl ginsenoside Rd (1) malonyl ginsenoside Rc (2) malonyl ginsenoside Rb2 (3) malonyl ginsenoside Rb1 (4) based on spectroscopic analyses including Nuclear magnetic resonance and HR-TOF/MS. The contents of malonyl ginsenoside Rb1 was highist as 5.44 mg/g of five years of ginseng. And malonyl ginsenoside Rd was lowest as 0.11 mg/g of six years of ginseng. Additionally, the malonyl ginsenoside Rd exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

고려인삼(Panax ginseng C.A. Meyer)을 70% EtOH 수용액으로 저온 추출한 뒤, 감압 농축한 추출물을SiO2, ODS column chromatograph 및 중압분취(MPLC) 장비를 반복 실시하여 4종의 인삼 사포닌 화합물을 분리 및 정제하였다. NMR 및 고분해능 질량분석 장비를 이용하여 malonyl ginsenoside Rd (1), Rc (2), Rb2 (3), 및 Rb1 (4)로 구조 동정하였다. 분리한 4종의 화합물에 대하여 UPLC-MS/MS 질량분석기를 이용하여 수삼의 5년 및 6년근 뿌리의 동체를 정량분석 하였으며, malonyl ginsenoside의 총 함량의 합은 각각 6.62 및 2.34 mg/g으로 5년근이 약 2.8배 높은 것을 확인하였다. 인삼으로부터 분리된 화합물 중 malonyl ginsenoside Rd의 경우, 알코올에 의해 저해된 HepG2세포에 대해서 간세포를 보호하는 효과가 있음을 확인하였다.

Keywords

References

  1. Gillis CN (1997) Panax ginseng pharmacology: A nitric oxide link. Biochem Pharmacol 54(1): 1-8 https://doi.org/10.1016/S0006-2952(97)00193-7
  2. Yun TK (2001) Brief introduction of Panax ginseng C.A. Meyer. J Korean Med Sci 16: S3-S5 https://doi.org/10.3346/jkms.2001.16.s.s3
  3. Park JD, Rhee DK, Lee YH (2005) Biological activites and chemistry of saponins from Panax ginseng C. A. Meyer. Phytochem Rev 4: 159-175 https://doi.org/10.1007/s11101-005-2835-8
  4. Lee JW, Ji SH, Choi BR, Choi DJ, Lee YG, Kim HG, Kim GS, Kim KI, Lee YH, Baek NI, Lee DY (2018) UPLC-QTOF/MS-based metabolomics applied for the quality evaluation of four processed Panax ginseng products. Molecules 23: 2062 https://doi.org/10.3390/molecules23082062
  5. Kitagawa I, Taniyama T, Yoshikawa M, Ikenishi Y, Nakagawa Y (1989) Chemical studies on crude drug processing. VI.: Chemical structures of malonyl-ginsenoside Rb1, Rb2, Rc, and Rd isolated from root of Panax ginseng C. A. Meyer. Chem Pharm Bull 37: 2961-2970 https://doi.org/10.1248/cpb.37.2961
  6. Shin JY, Choi EH, Wee JJ (2001) The difference of ginsenoside compositions according to the conditions of extraction and fractionation of crude ginseng saponins. Environ Toxicol Pharmacol 33: 282-287
  7. Kitagawa I, Taniyama T, Hayashi T, Yoshikawa M (1983) Malonylginsenosides Rb1, Rb2, Rc, and Rd, four new malonylated dammaranetype triterpene oligoglycosides from Ginseng Radix. Chem Pharm Bull 31(9): 3353-3356 https://doi.org/10.1248/cpb.31.3353
  8. Qiu S, Yang WZ, Yao CL, Shi XJ, Li JY, Lou Y, Duan YN, Wu WY, Guo DA (2017) Malonylginsenosides with potential antidiabetic activities from the flower buds of Panax ginseng. J Nat Prod 80(4): 899-908 https://doi.org/10.1021/acs.jnatprod.6b00789
  9. Liu Z, Li W, Li X, Zhang M, Chen L, Zheng YN, Sun GZ, Ruan CC (2013) Antidiabetic effects of malonylginsenosides from Panax ginseng on type 2 diabetic rats induced by high-fat diet and streptozotocin. J Ethnopharmacol 145: 233-240 https://doi.org/10.1016/j.jep.2012.10.058
  10. Liu Z, Li Y, Li X, Ruan CC, Wang LJ, Sun GZ (2012) The effects of dynamic changes of malonyl ginsenoside on evaluation and quality control of Panax ginseng C.A. Meyer. J Pharm Biomed Anal 64-65: 56-63 https://doi.org/10.1016/j.jpba.2012.02.005
  11. Wu W, Sun L, Zhang Z, Guo Y, Liu S (2015) Profiling and multivariate statistical analysis of Panax ginseng based on ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry. J Pharm Biomed Anal 107: 141-150 https://doi.org/10.1016/j.jpba.2014.12.030
  12. Nam KY, Hwang GB, Lee DY, Han JS, Noh HJ, Kim DH, Kim GS (2018) Variation in content of marker compounds and quality of processed white ginseng of different cultivation ages from different regions. Korean J Medicinal Crop Sci 26(5): 408-416 https://doi.org/10.7783/KJMCS.2018.26.5.408
  13. Yoon DH, Choi BR, Ma SH, Lee JW, Jo IH, Lee YS, Kim GS, Kim SM, Lee DY (2019) Metabolomics for age discrimination of ginseng using a multiplex approach to HR-MAS NMR spectroscopy, UPLC-QTOF/MS, and GC X GC-TOF/MS. Molecules 24(13): 2381 https://doi.org/10.3390/molecules24132381
  14. Philip S, Ritsa S, Dominic S, Anne M, James M, David V, Jonathan T, Heidi B, Susan K, Michael R (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Nat Cancer Inst 82(13): 1107-1112 https://doi.org/10.1093/jnci/82.13.1107
  15. Shen R, Laval S, Cao X, Yu B (2018) Synthesis of ${\Delta}{20}$-ginsenosides Rh4, (20E)-$Rh_{3}$, $Rg_{6}$, and $Rk_{1}$: a general approach to access dehydrated ginsenosides. J Org Chem 83(5): 2601-2610 https://doi.org/10.1021/acs.joc.7b02987
  16. Kim HG, Oh HJ, Ko JH, Song HS, Lee YG, Kang SC, Lee DY, Baek NI (2019) Lanceoleins A-G, hydroxychalcones, from the flowers of Coreopsis lanceolata and their chemopreventive effects against human colon cancer cells. Bioorg Chem 85: 274-281 https://doi.org/10.1016/j.bioorg.2019.01.003
  17. Peng M, Yi YX, Zhang D, Ding Y, Le J (2018) Stereoisomers of saponins in Panax notoginseng (Sanqi): a review front. Pharmacol 9(188): 1-18
  18. Sun GZ, Liu Z, Li XG, Zheng YN, Wang JY (2005) Isolation and identification of two malonylginsenosides from the fresh root of Panax ginseng. Chin J Anal Chem 33: 1783-1786 https://doi.org/10.3321/j.issn:0253-3820.2005.12.030
  19. Ruan CC, Liu Z, Li X, Liu X, Wang LJ, Pan HY, Zheng YN, Sun GZ, Zhang YS, Zhang LX (2010) Isolation and characterization of a new ginsenoside from the fresh root of Panax ginseng. Molecules 15: 2319-2325 https://doi.org/10.3390/molecules15042319
  20. Wan JY, Fan Y, Yu QT, Ge YZ, Yan CP, Raphael N, Li P, Ma ZH, Qi LW (2015) Inergrated evaluation of malonyl ginsenosides, amino acids and polysaccharides in fresh and processed ginseng. J Pharm Biomed Anal 107: 89-97 https://doi.org/10.1016/j.jpba.2014.11.014
  21. Liu Z, Wang LJ, Li X, Hu JN, Chen Y, Ruan CC, Sun GZ (2009) Hypoglycemic effects of malonyl-ginsenosides extracted from roots of Panax ginseng on Streptozotocin-induced diabetic mice. J Phytother 23: 1426-1430 https://doi.org/10.1002/ptr.2796