350 rub
Journal №9 for 2014 г.
Article in number:
Application of HPLC method with tanded mass spectrometry for identification ginsenosides in phytoextracts
Keywords: phytoadaptogene ginsenoside electro spray limits of detection
Authors:
I.V. Kazeev - Ph.D. (Eng.), Research Scientist, Russian Cancer Research Center of N.N. Blokhin Russian Academy of Medical Sciences, Moscow, Russia. E-mail: Ilya_delta@mail.ru
O.A. Bocharova - Dr. Sc. (Biol.), Chief of Laboratory, Russian Cancer Research Center of N.N. Blokhin Russian Academy of Medical Sciences, Moscow, Russia
V.E. Shevchenko - Dr. Sc. (Chem.), Chief of Laboratory, Russian Cancer Research Center of N.N. Blokhin Russian Academy of Medical Sciences, Moscow, Russia
E.V. Bocharov - Ph.D. (Med.), Senior Research Scientist, Russian Cancer Research Center of N.N. Blokhin Russian Academy of Medical Sciences; Institute of General Pathology and Pathophysiology RAMS
R.V. Karpova - Ph.D. (Biol.), Senior Research Scientist, Russian Cancer Research Center of N.N. Blokhin Russian Academy of Medical Sciences, Moscow, Russia
V.G. Kucheryany - Dr.Sc. (Med.), Chief Research Scientist, Institute of General Pathology and Pathophysiology RAMS, Moscow, Russia
V.A. Bykov - Academician RAS, Dr.Sc. (Eng.), Professor, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants, Moscow, Russia
Abstract:
Composition of phytoadaptogen Phytomix-40 (PhM-40) was developed at Blokhin Russian Cancer Research Center for prevention and comprehensive treatment of cancer. Numerous experimental and clinical studies demonstrated antimutagenic, antitumor, radioprotective, antioxidant, neuroprotective, immunomodulatory, including anti-inflammatory effects of PhM 40. Treatment effects on benign prostatic hyperplasia (age-related disease), leukoplakia of the oral mucosa (pre-cancer), advanced gastric cancer (IV stage of development) and Parkinson's disease (age-related neurodegenerative disease) were proven in the clinical studies of PhM-40. In the previous studies the presence of polyphenolic compounds and amino acids were determined in PhM 40 by reversed-phase high performance liquid chromatography with UV detection using standard samples. As a result, eight phenolic substances were found: hyperoside, rutin, naringenin, quercetin, luteolin, apigenin (flavonoids), arbutin (fenologlikozid) and glicirrizinic acid (triterpenoid saponins). Furthermore, 18 amino acids were identified, seven of them are essential. In this study analysis was carried out on a mass spectrometer TSQ Vantage which was connected to the HPLC Accela chromatograph column ACQUITY UPLC BEH C18 (1,7 micron, 2.1 x 100 mm, Waters). The triple quadrupole mass spectrometer Series Thermo Scientific TSQ was used. Were used ginsenosides standards Rg1, Rc, Rb1, Rb2, Rd, Rh1 and Rh2 (ChromaDex), Lanatozid C standard, water and methanol for HPLC (Sigma-Aldrich), and acetonitrile for HPLC (Reiden-deHaen). As a result of retention times and mass spectra comparison with standards, ginsenosides Rg1, Rc, Rb1, Rb2 and Rd (the same as in the ginseng extract) were identified in PhM 40. Thus, HPLC coupled with tandem mass spectrometry (HPLC-MS/MS) was successfully used in the present studies for identification and quantification of ginsenosides as components of phytopreparation (for example PhM 40).So, the approaches to standardization of herbal remedies consisted from the plant extracts components including ginsenosides were determined.
Pages: 22-33
References

  1. Bocharova O.A., Bary'shnikov A.Ju., Davy'dov M.I. Fitoadaptogeny' v onkologii i gerontologii. M.: MIA 2008. 218 c.
  2. Bocharova O.A., M.I. Davy'dov, Bary'shnikov A.Ju., Klimenkov A.A., Matveev V.B., Pozhariczkaya M.M., Ivanova-Smolenskaya I.A., Karpova R.V., Gorozhanskaya E'.G., Shejchenko O.P., Suxanov B.P., Kry'zhanovskij G.N., Bochkov N.P., By'kov V.A., Tutel'yan V.A. Kompleksny'e fitoadaptogeny' v onkologii i gerontologii // Vestnik RAMN. 2009. № 8. S. 21-25.
  3. Bocharov E.V.. Kucheryanu V.G., Bocharova O.A., Karpova R.V. Nejroprotektorny'e svojstva fitoadaptogenov // Vestnik RAMN. 2008. № 4. S. 47-50.
  4. Bocharova O.A., Karpova R.V., Golubeva V.A., Kasatkina N.N., Kurennaya O.N., Bocharov E.V. Toksikologicheskoe izuchenie preparata Fitomiks-40 // Gigiena i sanitariya. 1999. № 5. S. 60-61.
  5. Bocharova O.A., Karpova R.V., Il'enko V.A., Bocharov E.V., Kazeev I.V. Lejkoczitarny'e integriny' pri gepatokanczerogeneze my'shej vy'sokorakovoj linii SVA // Rossijskij bioterapevticheskij zhurnal. 2013. T. 12. № 3. S. 53-56.
  6. Bocharova O.A., Ly'zhenkova M.A., Mezenczeva M.V., Semernina V.V., Knyazhev V.A. Fitoadaptogen dlya profilakticheskoj onkologii: immunobiologicheskie kriterii sostava // Byulletenn' e'ksperimental'noj biologii i medicziny'. 2003. T. 136. № 12. S. 670-673.
  7. Bocharova O.A., Chekalina T.L., Mezenczeva M.V., Ly'zhenkova M.A., Kasatkina N.N., Pozhariczkaya M.M. Perspektivy' ispol'zovaniya Fitomiks-40 dlya profilakticheskoj onkologii // Rossijskij bioterapevticheskij zhurnal. 2002. T. 1. № 2. S. 114.
  8. Kurennaya O.N., Karpova R.V., Bocharova O.A., Kazeev I.V., Bocharov E.V., Korolev V.G. Antimutagenez mul'tifitoadaptogena v kletkax drozhzhej-saxaromiczetov // Genetika. 2013. T. 49. № 12. S. 1364-1369.
  9. Bocharov E.V., Karpova R.V., Kazeev I.V., Kucheryanu V.G., Bocharova O A. Issledovanie radiozashhitnoj aktivnosti mul'tifitoadaptogena v e'ksperimente na my'shax // Patologicheskaya fiziologiya i e'ksperimental'naya terapiya. 2013. № 3. S. 55-58.
  10. Karpova R.V., Bocharov E.V., Kazeev I.V., Kucheryanu V.G., Bocharova O.A. Radiozashhitnaya e'ffektivnost' mul'tifitoadaptogena v opy'tax na sobakax // Patologicheskaya fiziologiya i e'ksperimental'naya terapiya. 2013. № 4. S. 51-54.
  11. Bocharova O.A., Pozhariczkaya M.M., Chekalina T.L., Voronin V.F. Sovremenny'e aspekty' patogeneza i lecheniya lejkoplakii slizistoj obolochki polosti rta. Metodicheskoe rukovodstvo dlya vrachej. 2004. M. GOU VUNMC.
  12. Bocharova O.A., Pozhariczkaya M.M., Chekalina T.L., Ly'zhenkova M.A., Karpova R.V., Mezenczeva M.V., Ershov F.I. Lejkoplakiya slizistoj obolochki polosti rta: patogenez i vozmozhnosti korrekczii fitoadaptogenom // Byulleten' e'ksperimental'noj biologii i medicziny'. 2004. T. 138. № 12. S. 652-657.
  13. Bochkov N.P., Bocharova O.A., Aksenov A.A., Gorozhanskaya E'.G., Matveev V.B., Karpova R.V., Katosova L.D., Kosyakova N.V., Platonova V.I., Chebotarev A.N. Chastota xromosomny'x aberraczij v limfoczitax paczientov s dobrokachestvennoj giperplaziej predstatel'noj zhelezy' // Mediczinskaya genetika. 2005. T. 4. № 1. S. 15-19.
  14. Bocharova O.A., Davy'dov M.I., Klimenkov A.A., Bary'shnikov A.Ju., Karpova R.V., Chulkova S.V., Gorozhanskaya E'.G., Il'enko V.A. Perspektivy' primeneniya fitoadaptogena v lechenii rasprostranennogo raka zheludka // Byulleten' e'ksperimental'noj biologii i medicziny'. 2009. T. 148. № 7. S. 96-99.
  15. Bocharov E.V., Ivanova-Smolenskaya I.A., Poleshhuk V.V., Kucheryanu V.G., Il'enko V.A., Bocharova O.A. Vozmozhnosti fitoadaptogena-nejroprotektora pri lechenii nejrodegenerativnogo zabolevaniya (na primere bolezni Parkinsona) // Byulleten' e'ksperimental'noj biologii i medicziny'. 2010. T. 149. № 6. S. 619-621.
  16. Bocharova O.A., Matveev V.B., Karpova R.V., Aksenov A.A., Gorozhanskaya E'.G., Chebotarev A.N., Katosova L.D., Platonova V.I., Bochkov N.P. Korrekcziya klinicheskix i immunobiologicheskix pokazatelej u muzhchin s dobrokachestvennoj giperplaziej predstatel'noj zhelezy' fitoadaptogenom // Byulleten' e'ksperimental'noj biologii i medicziny'. 2006. T. 141. № 5. S. 555-559.
  17. Zenkevich I.G., Bagirova V.L., Sokol'skaya T.A., Nechaeva E.B. Obzor fiziko-ximicheskix metodov standartizaczii nastoek, e'kstraktov i e'liksirov v vedushhix stranax Evropy' i Ameriki // Farmacziya. 2002. № 1. S. 43-45.
  18. Bagirova V.L., Severczev V.A. Nastojki, e'kstrakty' i ix standartizacziya. Sankt-Peterburg. 2001.
  19. Yang W.Z., Ye M., Qiao X., Liu C.F., Miao W., Bo T., Tao H.Y., Guo D.A. A strategy for efficient discovery of new natural compounds by integrating orthogonal column chromatography and liquid chromatography/mass spectrometry analysis: Its application in Panax ginseng, Panax quinquefolium and Panax notoginseng to characterize 437 potential new ginsenosides // Anal. Chim. Acta. 2012. № 739. P. 56-66.
  20. Shejchenko V.I., Bocharova O.A., Shejchenko O.P., Bocharov E.V., Bary'shnikov Ju.A., By'kov V.A. Analiticheskie vozmozhnosti metoda JaMR dlya opredeleniya komponentov preparata Fitomiks-40 // Zavodskaya laboratoriya. Diagnostika materialov. 2006. T. 72. № 8. S. 15-23.
  21. Shejchenko O.P., Bocharova O.A., Shejchenko V.I., Tolkachev O.N., Bocharov E.V., Karpova R.V., By'kov V.A. Vozmozhnost' ispol'zovaniya e'lektronny'x spektrov pogloshheniya dlya standartizaczii mnogokomponentnogo preparata «Fitomiks-40» // Voprosy' biologicheskoj, mediczinskoj i farmaczevticheskoj ximii. 2007. № 2. S. 20-25.
  22. Shejchenko O.P., Bocharova O.A., Krapivkin B.A., Uyutova E.V., Tolkachev O.N., Bocharov E.V., By'kov V.A. Opredelenie ximicheskogo sostava letuchix soedinenij fitoadaptogena «Fitomiks-40» metodom xromatomass-spektrometrii // Voprosy' biologicheskoj, mediczinskoj i farmaczevticheskoj ximii. 2008. № 5. S. 18-28.
  23. Shejchenko O.P., Bocharova O.A., Krapivkin B.A., Uyutova E.V., Karpova R.V., Kazeev I.V., Bocharov E.V., By'kov V.A. Issledovanie kompleksnogo fitoadaptogena metodom VE'ZhX // Voprosy' biologicheskoj, mediczinskoj i farmaczevticheskoj ximii. 2012. № 10. S. 52-59.
  24. Patent № 2099410 (RF). Kompozicziya ingredientov dlya preparata Fitomiks-40 / O.A. Bocharova. 1998.
  25. Liu Z., Li Y., Li X., Ruan C.C., Wang L.J., Sun G.Z. The effects of dynamic changes of malonyl ginsenosides on evaluation and quality control of Panax ginseng C.A. Meyer // J. Pharm. Biomed. Anal. 2012. V. 64-65. P. 56-63.
  26. Wong V.K., Cheung S.S., Li T., Jiang Z.H., Wang J.R., Dong H., Yi X.Q., Zhou H., Liu L. Asian ginseng extract inhibits in vitro and in vivo growth of mouse lewis lung carcinoma via modulation of ERK-p53 and NF-κB signaling // J. Cell Biochem. 2010. V. 111. № 4. P. 899-910.
  27. Morinaga O., Uto T., Yuan C.S., Tanaka H., Shoyama Y. Evaluation of a new eastern blotting technique for the analysis of ginsenoside Re in American ginseng berry pulp extracts // Fitoterapia. 2010. V. 81. № 4. - P. 284-288.
  28. Li H., Lee J.H., Ha J.M. Effective purification of ginsenosides from cultured wild ginseng roots, red ginseng, and white ginseng with macroporous resins // J. Microbiol. Biotechnol.  2008. V. 18. № 11. P. 1789-1791.
  29. Zhao Q., Zheng X., Jiang J., Zhou H., Hu P. Determination of ginsenoside Rg3 in human plasma and urine by high performance liquid chromatography-tandem mass spectrometry // J. Chromatogr B Analyt. Technol. Biomed. Life Sci. 2010. V. 878. № 24. P. 2266-2273.
  30. Li X., Tang M.H., Zhang F., Yuan Z., Chen L.J., Wang X.H. Quantitative determination of ginsenoside Re and Rg1 in human plasma by ultra performance liquid chromatography-tandem mass spectrometry // Sichuan Da Xue Xue Bao Yi Xue Ban. 2009. V. 40. № 5. P. 912-917.
  31. Deng G.F., Wang D.L., Meng M.X., Hu F., Yao T.W. Simultaneous determination of notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and icariin in rat plasma by ultra-performance liquid chromatography-tandem mass spectrometry // J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2009. V. 877. № 22. P. 2113-2122.
  32. Chen G., Yang M., Song Y., Lu Z., Zhang J., Huang H., Guan S., Wu L., Guo D.A. Comparative analysis on microbial and rat metabolism of ginsenoside Rb1 by high-performance liquid chromatography coupled with tandem mass spectrometry // Biomed. Chromatogr. 2008. V. 22. № 7. P. 779-785.