Analytical and Bioanalytical Chemistry

, Volume 405, Issue 13, pp 4511–4522 | Cite as

Simplified ultrasonically- and microwave-assisted solvent extractions for the determination of ginsenosides in powdered Panax ginseng rhizomes using liquid chromatography with UV absorbance or electrospray mass spectrometric detection

Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

New approaches for the recovery of ginsenosides are presented that greatly simplify the liquid chromatographic (LC) determination of the total content of eight ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, Rg1 and Rg2) in powdered Panax ginseng rhizomes. The extraction protocols not only recover the neutral ginsenosides, but also simultaneously incorporate base-catalyzed hydrolysis of the malonyl-ginsenosides using dilute potassium hydroxide added to the methanol–water extractant. This eliminates the need for an independent extraction step followed by acid- or base-catalyzed hydrolysis. Both ultrasonically-assisted and microwave-assisted extraction methods are developed. The optimization of these simplified methods to remove pendant malonate esters, while retaining the glycosidic linkages, was determined by LC through variation of the extraction/hydrolysis time, order of hydrolysis reagent addition, and evaluation of multiple extractions. A comparison of the ginsenoside profiles obtained with and without addition of base to the extractant solution was made using LCMS with positive-mode electrospray ionization (ESI+) detection. A number of malonyl-ginsenosides were tentatively identified by their mass spectral fragmentation spectra and indicating that they were converted to the free ginsenosides by the new extraction/hydrolysis procedure.

Figure

LCUV chromatograms for different extraction solvents

Keywords

Ginsenoside Liquid Chromatography with Ultraviolet/Visible Detection (LCUV) LC with Electrospray Mass Spectrometry (LCMS) Panax ginseng Malonyl-ginsenoside Ultrasonically-assisted solvent extraction Microwave-assisted solvent extraction 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

This research was performed while the author CM White held a National Research Council Research Associateship Award at the National Institute of Standards and Technology. Partial funding for this work was provided by the National Institutes of Health, Office of Dietary Supplements.

Supplementary material

216_2013_6871_MOESM1_ESM.pdf (1.1 mb)
ESM 1 (PDF 1123 kb)

References

  1. 1.
    Zhou W, Chai H, Lin PH, Lumsden AB, Yao Q, Chen CJ (2004) Molecular mechanisms and clinical applications of ginseng root for cardiovascular disease. Med Sci Monit 10:RA187–RA192Google Scholar
  2. 2.
    Lee KY, Lee YH, Kim S (1997) Ginsenoside-Rg5 suppresses cyclin E-dependent protein kinase activity via up-regulation of p21 Waf1 with concomitant down-regulation of cdc25A in SK-HEP-1 cells. Anticancer Res 17:1067–1072Google Scholar
  3. 3.
    Sotaniemi EA, Haapakoski E, Rautio A (1995) Ginseng therapy in non-lnsulin-dependent, diabetic patients. Diabetes Care 18:1373–1375CrossRefGoogle Scholar
  4. 4.
    Lewis R, Wake G, Court G, Court JA, Pickering AT, Kim YC, Perry EK (1999) Non-ginsenoside nicotinic activity in ginseng species. Phytother Res 13:59–64CrossRefGoogle Scholar
  5. 5.
    Yin J, Zhang H, Ye J (2008) Traditional Chinese medicine in treatment of metabolic syndrome. Endocr Metab Immune Disord Drug Targets 8(2):99–111CrossRefGoogle Scholar
  6. 6.
    Xiang YZ, Shang HW, Gao XM, Zhang BL (2008) A comparison of the ancient use of ginseng in traditional Chinese medicine with modern pharmacological experiments and clinical trials. Phytother Res 22:851–858CrossRefGoogle Scholar
  7. 7.
    Panossian A, Wikman G (2009) Evidence-based efficacy of adaptogens in fatigue, and molecular mechanisms related to their stress-protective activity. Curr Clin Pharmacol 4:198–219CrossRefGoogle Scholar
  8. 8.
    Sugimoto S, Nakamura S, Matsuda H, Kitagawa N, Yoshikawa M (2009) Chemical constituents from seeds of Panax ginseng: structure of new dammarane-type triterpene ketone, panaxadione, and HPLC comparisons of seeds and flesh. Chem Pharm Bull 57:283–287CrossRefGoogle Scholar
  9. 9.
    Liu Z, Li Y, Ruan CC, Wand LJ, Sun GZ (2012) The effects of dynamic changes of malonyl ginsenosides on the evaluation and quality control of Panax ginseng C.A. Meyer. J Pharm Biomed Anal 64–65:56–63CrossRefGoogle Scholar
  10. 10.
    Teng R, Li H, Chem J, Wang D, He Y, Yang C (2002) Complete assignment of 1H and 13C NMR data for nine protopanaxatriol glycosides. Magn Reson Chem 40:483–488CrossRefGoogle Scholar
  11. 11.
    Kang J, Lee S, Kang S, Kwon HN, Park JH, Kwon SW, Park S (2008) NMR-based metabolomics approach for the differentiation of ginseng (Panax ginseng) roots from different origins. Arch Pharm Res 31:330–336CrossRefGoogle Scholar
  12. 12.
    Wu W, Song F, Guo D, Mi J, Qin Q, Yu Q, Liu S (2012) Mass spectrometry-based approach in ginseng research: a promising way to metabolomics. Curr Anal Chem 8:43–66CrossRefGoogle Scholar
  13. 13.
    Sloley BD, Ridgway D, Semple HA, Tam YK, Coutts RT, Lodenberg R, Tam-Zaman N (2006) A method for the analysis of ginsenosides, malonyl ginsenosides, and hydrolyzed ginsenosides using high-performance liquid chromatography with ultraviolet and positive mode electrospray ionization mass spectrometric detection. J AOAC Intern 89:16–21Google Scholar
  14. 14.
    Xing Q, Liang T, Shen G, Wang X, Yu J (2012) Comprehensive HILIC x RPLC with mass spectrometry detection for the analysis of saponins in Panax notoginseng. Analyst 137:2239–2249CrossRefGoogle Scholar
  15. 15.
    Court W, Hendel J, Elmi J (1996) Reversed-phase high-performance liquid chromatographic determination of Ginsenosides in Panax quinquefolium. J Chrom A 755:11–17CrossRefGoogle Scholar
  16. 16.
    Shi Y, Sun C, Zheng Y, Wang Y (2010) Simultaneous determination of nine ginsenosides in functional foods by high performance liquid chromatography with diode array detector detection. Food Chem 123:1322–1327CrossRefGoogle Scholar
  17. 17.
    Lee S-I, Kwon H-J, Lee Y-M, Lee J-H, Hong S-P (2012) Simultaneous analysis method for polar and non-polar ginsenosides in red ginseng by reversed-phase HPLE-PAD. J Pharm Biomed Anal 60:80–85CrossRefGoogle Scholar
  18. 18.
    Nolte J, Nigge W, Hohaus E, Cho I-H, Lentz H (2005) Comparative study of extraction techniques with ammonia or methanol/water for the isolation of ginsenosides using HPLC/MS. Z Naturforsch 60b:205–210Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg (outside the USA) 2013

Authors and Affiliations

  1. 1.Chemical Sciences DivisionNational Institute of Standards and TechnologyGaithersburgUSA

Personalised recommendations