DOI QR코드

DOI QR Code

Simultaneous determination of 30 ginsenosides in Panax ginseng preparations using ultra performance liquid chromatography

  • Park, Hee-Won (Korea Ginseng Corporation Research Institute, Korea Ginseng Corporation) ;
  • In, Gyo (Korea Ginseng Corporation Research Institute, Korea Ginseng Corporation) ;
  • Han, Sung-Tai (Korea Ginseng Corporation Research Institute, Korea Ginseng Corporation) ;
  • Lee, Myoung-Woo (Korea Ginseng Corporation Research Institute, Korea Ginseng Corporation) ;
  • Kim, So-Young (Waters Korea Limited, KICOX Venture Center) ;
  • Kim, Kyung-Tack (Processing Technology Research Group, Korea Food Research Institute) ;
  • Cho, Byung-Goo (Korea Ginseng Corporation Research Institute, Korea Ginseng Corporation) ;
  • Han, Gyeong-Ho (Korea Ginseng Corporation Research Institute, Korea Ginseng Corporation) ;
  • Chang, Il-Moo (Korea Ginseng Corporation Research Institute, Korea Ginseng Corporation)
  • Received : 2013.05.13
  • Accepted : 2013.06.28
  • Published : 2013.10.15

Abstract

A quick and simple method for simultaneous determination of the 30 ginsenosides (ginsenoside Ro, Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, 20(S)-Rg2, 20(R)-Rg2, 20(S)-Rg3, 20(R)-Rg3, 20(S)-Rh1, 20(S)-Rh2, 20(R)-Rh2, F1, F2, F4, Ra1, Rg6, Rh4, Rk3, Rg5, Rk1, Rb3, Rk2, Rh3, compound Y, compound K, and notoginsenoside R1) in Panax ginseng preparations was developed and validated by an ultra performance liquid chromatography photo diode array detector. The separation of the 30 ginsenosides was efficiently undertaken on the Acquity BEH C-18 column with gradient elution with phosphoric acids. Especially the chromatogram of the ginsenoside Ro was dramatically enhanced by adding phosphoric acid. Under optimized conditions, the detection limits were 0.4 to 1.7 mg/L and the calibration curves of the peak areas for the 30 ginsenosides were linear over three orders of magnitude with a correlation coefficients greater than 0.999. The accuracy of the method was tested by a recovery measurement of the spiked samples which yielded good results of 89% to 118%. From these overall results, the proposed method may be helpful in the development and quality of P. ginseng preparations because of its wide range of applications due to the simultaneous analysis of many kinds of ginsenosides.

Keywords

References

  1. Soldati F. Panax ginseng: standardization and biological activity. In: Cutler SJ, Cutler HG, eds. Biologically active natural products. New York: CRC press, 2000. p.209-232.
  2. Christensen LP. Ginsenosides chemistry, biosynthesis, analysis, and potential health effects. Adv Food Nutr Res 2009;55:1-99.
  3. Fuzzati N. Analysis methods of ginsenosides. J Chromatogr B Analyt Technol Biomed Life Sci 2004;812:119-133. https://doi.org/10.1016/j.jchromb.2004.07.039
  4. Oleszek WA. Chromatographic determination of plant saponins. J Chromatogr A 2002;967:147-162. https://doi.org/10.1016/S0021-9673(01)01556-4
  5. Oleszek W, Bialy Z. Chromatographic determination of plant saponins: an update (2002-2005). J Chromatogr A 2006;1112:78-91. https://doi.org/10.1016/j.chroma.2006.01.037
  6. Sanada S, Kondo N, Shoji J, Tanaka O, Shibata S. Studies on the saponins of ginseng. I. Structures of ginsenoside-Ro, -Rb1, -Rb2, -Rc and -Rd. Chem Pharm Bull (Tokyo) 1974;22:421-428. https://doi.org/10.1248/cpb.22.421
  7. Bombardelli E, Bonati A, Gabetta B, Martinelli EM. Gas-liquid chromatographic method for determination of ginsenosides in Panax ginseng. J Chromatogr A 1980;196:121-132. https://doi.org/10.1016/S0021-9673(00)80364-7
  8. Cui JF. Identification and quantification of ginsenosides in various commercial ginseng preparations. Eur J Pharm Sci 1995;3:77-85. https://doi.org/10.1016/0928-0987(94)00077-D
  9. Nishi H, Terabe S. Micellar electrokinetic chromatography perspectives in drug analysis. J Chromatogr A 1996;735:3-27. https://doi.org/10.1016/0021-9673(95)01381-4
  10. Yap KY, Chan SY, Lim CS. Infrared-based protocol for the identification and categorization of ginseng and its products. Food Res Int 2007;40:643-652. https://doi.org/10.1016/j.foodres.2006.11.009
  11. Jung DW, Lee JM, Sung CK. Enzyme-linked immunosorbent assay for the determination of 20(S)-protopanaxatriol. Anal Chim Acta 2002;462:157-163. https://doi.org/10.1016/S0003-2670(02)00340-9
  12. Yamaguchi H, Kasai R, Matsuura H, Tanaka O, Fuwa T. High-performance liquid chromatographic analysis of acidic saponins of ginseng and related plants. Chem Pharm Bull (Tokyo) 1988;36:3468-3473. https://doi.org/10.1248/cpb.36.3468
  13. Kanazawa H, Nagata Y, Matsushima Y, Tomoda M, Takai N. Simultaneous determination of ginsenosides and saikosaponins by high-performance liquid chromatography. J Chromatogr 1990;507:327-332. https://doi.org/10.1016/S0021-9673(01)84210-2
  14. Kanazawa H, Nagata Y, Matsushima Y, Tomoda M, Takai N. Determination of acidic saponins in crude drugs by high-performance liquid chromatography on octadecylsilyl porous glass. J Chromatogr 1993;630:408-414. https://doi.org/10.1016/0021-9673(93)80479-R
  15. Chung WC, Sheu SJ. Determination of ginsenosides in ginseng crude extracts by high-performance liquid chromatography. J Chromatogr A 1994;685:243-251. https://doi.org/10.1016/0021-9673(94)00724-1
  16. Court WA, Hendel JG, Elmi J. Reversed-phase high-performance liquid chromatography determination of ginsenosides of Panax quinquefolium. J Chromatogr A 1996;755:11-17. https://doi.org/10.1016/S0021-9673(96)00580-8
  17. Shi Y, Sun C, Zheng B, Li Y, Wang Y. Simultaneous determination of nine ginsenosides in functional foods by high-performance liquid chromatography with diode array detector detection. Food Chem 2010;123:1322-1327. https://doi.org/10.1016/j.foodchem.2010.06.014
  18. Park MK, Park JH, Han SB, Shin YG, Park IH. High-performance liquid chromatographic analysis of ginseng saponins using evaporative light scattering detection. J Chromatogr A 1996;736:77-81. https://doi.org/10.1016/0021-9673(95)01323-7
  19. Kwon SW, Han SB, Park IH, Kim JM, Park MK, Park JH. Liquid chromatographic determination of less polar ginsenosides in processed ginseng. J Chromatogr A 2001;921:335-339. https://doi.org/10.1016/S0021-9673(01)00869-X
  20. Kim SN, Ha YW, Shin H, Son SH, Wu SJ, Kim YS. Simultaneous quantification of 14 ginsenosides in Panax ginseng C.A. Meyer (Korean red ginseng) by HPLCELSD and its application to quality control. J Pharm Biomed Anal 2007;45:164-170. https://doi.org/10.1016/j.jpba.2007.05.001
  21. Sun BS, Gu LJ, Fang ZM, Wang CY, Wang Z, Lee MR, Li Z, Li JJ, Sung CK. Simultaneous quantification of 19 ginsenosides in black ginseng developed from Panax ginseng by HPLC-ELSD. J Pharm Biomed Anal 2009;50:15-22. https://doi.org/10.1016/j.jpba.2009.03.025
  22. Shangguan D, Han H, Zhao R, Zhao Y, Xiong S, Liu G. New method for high-performance liquid chromatographic separation and fluorescence detection of ginsenosides. J Chromatogr A 2001;910:367-372. https://doi.org/10.1016/S0021-9673(00)01208-5
  23. Lee SI, Kwon HJ, Lee YM, Lee JH, Hong SP. Simultaneous analysis method for polar and non-polar ginsenosides in red ginseng by reversed-phase HPLC-PAD. J Pharm Biomed Anal 2012;60:80-85. https://doi.org/10.1016/j.jpba.2011.08.030
  24. Novakova L, Matysova L, Solich P. Advantages of application of UPLC in pharmaceutical analysis. Talanta 2006;68:908-918. https://doi.org/10.1016/j.talanta.2005.06.035
  25. Guan J, Lai CM, Li SP. A rapid method for the simultaneous determination of 11 saponins in Panax notoginseng using ultra performance liquid chromatography. J Pharm Biomed Anal 2007;44:996-1000. https://doi.org/10.1016/j.jpba.2007.03.032
  26. Wang X, Zhao T, Gao X, Dan M, Zhou M, Jia W. Simultaneous determination of 17 ginsenosides in rat urine by ultra performance liquid chromatography-mass spectrometry with solid-phase extraction. Anal Chim Acta 2007;594:265-273. https://doi.org/10.1016/j.aca.2007.05.032
  27. In G, Ahn NG, Bae BS, Han ST, Noh KB, Kim CS. New method for simultaneous quantification of 12 ginsenosides in red ginseng powder and extract: in-house method validation. J Ginseng Res 2012;36:205-210. https://doi.org/10.5142/jgr.2012.36.2.205
  28. Taverniers I, Loose MD, Bockstaele EV. Trends in quality in the analytical laboratory. II. Analytical method validation and quality assurance. TrAC Trends Anal Chem 2004;23:535-552. https://doi.org/10.1016/j.trac.2004.04.001
  29. Ermer J. Validation in pharmaceutical analysis. Part I: an integrated approach. J Pharm Biomed Anal 2001;24:755-767. https://doi.org/10.1016/S0731-7085(00)00530-6
  30. Wang Y, You J, Yu Y, Qu C, Zhang H, Ding L, Zhang H, Li X. Analysis of ginsenosides in Panax ginseng in high pressure microwave-assisted extraction. Food Chem 2008;110:161-167. https://doi.org/10.1016/j.foodchem.2008.01.028
  31. Ha YW, Lim SS, Ha IJ, Na YC, Seo JJ, Shin H, Son SH, Kim YS. Preparative isolation of four ginsenosides from Korean red ginseng (steam-treated Panax ginseng C. A. Meyer), by high-speed counter-current chromatography coupled with evaporative light scattering detection. J Chromatogr A 2007;1151:37-44. https://doi.org/10.1016/j.chroma.2007.01.038
  32. Lee YJ, Kim HY, Kang KS, Lee JG, Yokozawa T, Park JH. The chemical and hydroxyl radical scavenging activity changes of ginsenoside-Rb1 by heat processing. Bioorg Med Chem Lett 2008;18:4515-4520. https://doi.org/10.1016/j.bmcl.2008.07.056
  33. Yamabe N, Kim YJ, Lee S, Cho EJ, Park SH, Ham J, Kim HY, Kang KS. Increase in antioxidant and anticancer effects of ginsenoside Re-lysine mixture by Maillard reaction. Food Chem 2013;138:876-883. https://doi.org/10.1016/j.foodchem.2012.12.004

Cited by

  1. Simultaneous Determination of Original, Degraded Ginsenosides and Aglycones by Ultra High Performance Liquid Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry for Quantitative Evaluation of Du-Shen-Tang, the Decoction of Ginseng vol.19, pp.4, 2014, https://doi.org/10.3390/molecules19044083
  2. Autophagy-associated Targeting Pathways of Natural Products during Cancer Treatment vol.15, pp.24, 2014, https://doi.org/10.7314/APJCP.2014.15.24.10557
  3. An enzymatic protocol for absolute quantification of analogues: application to specific protopanoxadiol-type ginsenosides vol.17, pp.4, 2015, https://doi.org/10.1039/C5GC00091B
  4. Study vol.2015, pp.1741-4288, 2015, https://doi.org/10.1155/2015/817096
  5. Metabolomic Approach for Discrimination of Four- and Six-Year-Old Red Ginseng (Panax ginseng) Using UPLC-QToF-MS vol.64, pp.9, 2016, https://doi.org/10.1248/cpb.c16-00240
  6. Red ginseng and vitamin C increase immune cell activity and decrease lung inflammation induced by influenza A virus/H1N1 infection vol.68, pp.3, 2016, https://doi.org/10.1111/jphp.12529
  7. Antimelanogenesis Activity of Hydrolyzed Ginseng Extract (GINST) via Inhibition of JNK Mitogen-activated Protein Kinase in B16F10 Cells vol.81, pp.8, 2016, https://doi.org/10.1111/1750-3841.13380
  8. A Metabolomic Approach for the Discrimination of Red Ginseng Root Parts and Targeted Validation vol.22, pp.3, 2017, https://doi.org/10.3390/molecules22030471
  9. A new approach for authentication of four ginseng herbs and their related products based on the simultaneous quantification of 19 ginseng saponins by UHPLC-TOF/MS coupled with OPLS-DA vol.7, pp.74, 2017, https://doi.org/10.1039/C7RA06812C
  10. Neurogenic Traditional Chinese Medicine as a Promising Strategy for the Treatment of Alzheimer’s Disease vol.18, pp.2, 2017, https://doi.org/10.3390/ijms18020272
  11. Stereoisomers of Saponins in Panax notoginseng (Sanqi): A Review vol.9, pp.1663-9812, 2018, https://doi.org/10.3389/fphar.2018.00188
  12. Optimization of Fermentation Process Parameters for Ginsenoside Re Bioconversion by Plackett-Burman and Box-Benhnken Design vol.238, pp.2261-236X, 2018, https://doi.org/10.1051/matecconf/201823804001
  13. Protective Effects of Processed Ginseng and Its Active Ginsenosides on Cisplatin-Induced Nephrotoxicity: In Vitro and in Vivo Studies vol.63, pp.25, 2015, https://doi.org/10.1021/acs.jafc.5b00782
  14. Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia vol.6, pp.None, 2016, https://doi.org/10.1186/s13568-016-0277-x
  15. 인삼의 추출조건 및 진세노사이드의 HPLC 분석법 평가 vol.24, pp.1, 2013, https://doi.org/10.7783/kjmcs.2016.24.1.47
  16. Ginsenoside Rg3 Decreases Fibrotic and Invasive Nature of Endometriosis by Modulating miRNA-27b: In Vitro and In Vivo Studies vol.7, pp.None, 2013, https://doi.org/10.1038/s41598-017-17956-0
  17. Quantitative Analysis of Ginsenosides Using Relative Response Factors from Ginsenoside Rf vol.16, pp.2, 2013, https://doi.org/10.20402/ajbc.2017.0186
  18. Protective Effects and Target Network Analysis of Ginsenoside Rg1 in Cerebral Ischemia and Reperfusion Injury: A Comprehensive Overview of Experimental Studies vol.7, pp.12, 2013, https://doi.org/10.3390/cells7120270
  19. Red Ginseng Attenuates Aβ-Induced Mitochondrial Dysfunction and Aβ-mediated Pathology in an Animal Model of Alzheimer’s Disease vol.20, pp.12, 2019, https://doi.org/10.3390/ijms20123030
  20. High-density immobilization of a ginsenoside-transforming β-glucosidase for enhanced food-grade production of minor ginsenosides vol.103, pp.17, 2013, https://doi.org/10.1007/s00253-019-09951-4
  21. Recent Advances in Ginsenosides as Potential Therapeutics Against Breast Cancer vol.19, pp.25, 2013, https://doi.org/10.2174/1568026619666191018100848
  22. Red Ginseng Inhibits Tau Aggregation and Promotes Tau Dissociation In Vitro vol.2020, pp.None, 2020, https://doi.org/10.1155/2020/7829842
  23. 인삼의 열처리 과정 중 생성되는 3종의 수산화진세노사이드에 대한 연구 vol.51, pp.4, 2013, https://doi.org/10.22889/kjp.2020.51.4.255
  24. Effects of immersion in fermented tea liquid and steam treatments on physicochemical properties and ginsenoside profiles of Korean ginseng vol.45, pp.1, 2013, https://doi.org/10.1111/jfpp.15050
  25. Simultaneous Determination of 25 Ginsenosides by UPLC-HRMS via Quantitative Analysis of Multicomponents by Single Marker vol.2021, pp.None, 2013, https://doi.org/10.1155/2021/9986793
  26. Comparison of the Constituents of Processed Korean and American Ginseng Grown in Korea for Six Years vol.29, pp.1, 2021, https://doi.org/10.7783/kjmcs.2021.29.1.35
  27. Simultaneous Quantitative Analysis of Ginsenosides Isolated from the Fruit of Panax ginseng C.A. Meyer and Regulation of HO-1 Expression through EGFR Signaling Has Anti-Inflammatory and Osteogenic Ind vol.26, pp.7, 2013, https://doi.org/10.3390/molecules26072092
  28. Application of Identification and Evaluation Techniques for Ethnobotanical Medicinal Plant of Genus Panax: A Review vol.51, pp.4, 2013, https://doi.org/10.1080/10408347.2020.1736506
  29. Evaluation of the Saponin Content in Panax vietnamensis Acclimatized to Lam Dong Province by HPLC-UV/CAD vol.26, pp.17, 2013, https://doi.org/10.3390/molecules26175373
  30. Simultaneous quantitative assays of 15 ginsenosides from 119 batches of ginseng samples representing 12 traditional Chinese medicines by ultra-high performance liquid chromatography coupled with charg vol.1655, pp.None, 2013, https://doi.org/10.1016/j.chroma.2021.462504
  31. Isolation and Identification of Non-Conjugated Linoleic Acid from Processed Panax ginseng Using LC-MS/MS and 1H-NMR vol.8, pp.11, 2021, https://doi.org/10.3390/separations8110208
  32. Predominance of oleanane-type ginsenoside R0 and malonyl esters of protopanaxadiol-type ginsenosides in the 20-year-old suspension cell culture of Panax japonicus C.A. Meyer vol.177, pp.None, 2013, https://doi.org/10.1016/j.indcrop.2021.114417