HPLC-MS Analysis of Ginsenosides in Morphogenic Cell Cultures of Ginseng (Panax ginseng C.A. Meyer)

T. Yu. Gorpenchenko; Горпенченко Т. Ю.; V. P. Grigorchuk; Григорчук В. П.; V. V. Makhankov; Маханьков В. В.; D. V. Kochkin; Кочкин Д. В.; Yu. V. Vologzhanina; Вологжанина Ю. В.; M. T. Khandy; Ханды М. Т.; G. K. Chernoded; Чернодед Г. К.; Yu. N. Zhuravlev; Журавлев Ю. Н.

doi:10.31857/S0015330323601036

HPLC-MS Analysis of Ginsenosides in Morphogenic Cell Cultures of Ginseng (Panax ginseng C.A. Meyer)

Authors: Gorpenchenko T.Y.¹, Grigorchuk V.P.¹, Makhankov V.V.², Kochkin D.V.³^,4, Vologzhanina Y.V.¹, Khandy M.T.¹, Chernoded G.K.¹, Zhuravlev Y.N.¹
Affiliations:
1. Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch, Russian Academy of Sciences
2. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences
3. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences
4. Moscow State University
Issue: Vol 70, No 7 (2023)
Pages: 823-835
Section: ЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИ
URL: https://journals.rcsi.science/0015-3303/article/view/233788
DOI: https://doi.org/10.31857/S0015330323601036
EDN: https://elibrary.ru/ATLJPG
ID: 233788

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Abstract

The composition of triterpene glycosides in different in vitro tissues of cell lines (PgR–rhizogenic and PgG–hemogenic) and roots of ginseng plants (Panax ginseng C.A. Meyer), grown on plantations in Primorsky Krai (Russian Federation) was determined using the HPLC-MS method. More than 60 triterpene glycosides have been detected and characterized. The maximum content of ginsenosides was identified in the leaves of the hemogenic line. The total amount of ginsenosides in the PgG line embryonic tissue was lower compared to all other samples studied mainly due to the amount of protopanaxadiols. In all parts of the PgR line, the content of protopanaxatriol Re was two times higher than in the root of the intact plant. A high content of protopanaxatriols and oleanolic ginsenosides was found in the basal parts of PgR and PgG calli. Thus, morphogenic cell lines not only repeated the pattern of qualitative ginsenosides composition in comparison with individual parts of native roots and leaves but also demonstrated an increase in their diversity and quantity.

Keywords

Panax ginseng, ginsenosides, ginseng, plant cell culture, morphogenic culture, triterpene glycosides

References

Журавлев Ю.Н., Томских А.А., Горпенченко Т.Ю. История и перспективы генетических исследований женьшеня на Дальнем Востоке России // Вестник ДВО РАН. 2022. № 4. С. 101. https://doi.org/10.37102/0869-7698_2022_224_04_9
Chen W., Balan P., Popovich B.G. Ginsenosides analysis of New Zealand-grown forest Panax ginseng by LC-QTOF-MS/MS // J. Ginseng Res. 2020. V. 44. P. 552. https://doi.org/10.1016/j.jgr.2019.04.007
Yang Y., Ju Z., Yang Y., Zhang Y., Yang L., Wang Z. Phytochemical analysis of Panax species: a review // J. Ginseng Res. 2021. V. 45. P. 1. https://doi.org/10.1016/j.jgr.2019.12.009
Du Z., Li J., Zhang X., Pei J., Huang L. An integrated LC-MS-based strategy for the quality assessment and discrimination of three Panax species // Molecules. 2018. V. 23. P. 2988. https://doi.org/10.3390/molecules23112988
Li F., Lv C., Li Q., Wang J., Song D., Liu P., Zhang D., Lu J. Chemical and bioactive comparison of flowers of Panax ginseng Meyer, Panax quinquefolius L., and Panax notoginseng Burk // J. Ginseng Res. 2017. V. 41. P. 487. https://doi.org/10.1016/jjgr.2016.08.008
Lee J.W., Choi B.-R., Kim Y.-C., Choi D.J., Lee Y.-S., Kim G.-S., Baek N.-I., Kim S.-Y., Lee D.Y. Comprehensive profiling and quantification of ginsenosides in the root, stem, leaf, and berry of Panax ginseng by UPLC-QTOF/MS // Molecules. 2017. V. 22 P. 2147. https://doi.org/10.3390/molecules22122147
Lee J., Han H., Yuan X., Park E., Lee J., Kim J.H. A rapid, simultaneous and quantitative analysis of 26 ginsenosides in white and red Panax ginseng using LC-M-S/MS // Appl. Biol. Chem. 2021. V. 64:13. https://doi.org/10.1186/s13765-020-00588-w
Di P., Yan Y., Wang P., Yan M., Wang Y.-P., Huang L.-Q. Integrative SMRT sequencing and ginsenoside profiling analysis provide insights into the biosynthesis of ginsenoside in Panax quinquefolium // Chin. J. Nat. Med. 2022. V. 20. P. 614. https://doi.org/10.1016/S1875-5364(22)60198-5
Gantait S., Mitra M., Chen J.-T. Biotechnological interventions for ginsenosides production // Biomolecules. 2020. V. 10. P. 538. https://doi.org/10.3390/biom10040538
Xu F., Valappil A.K., Mathiyalagan R., Tran T.N.A., Ramadhania Z.M., Awais M., Yang D.C. In vitro cultivation and ginsenosides accumulation in Panax ginseng: a review // Plants. 2023. V. 12. P. 3165. https://doi.org/10.3390/ plants12173165
Nosov A.M., Popova E.V., Kochkin D.V. Isoprenoid production via plant cell cultures: biosynthesis, accumulation and scaling-up to bioreactors // Production of biomass and bioactive compounds using bioreactor technology / Ed. Paek K.Y., Murthy H., Zhong J.J. Dordrecht: Springer-Verlag. 2014. P. 563. https://doi.org/10.1007/978-94-017-9223-3_23
Atanasov A.G., Waltenberger B., Pferschy-Wenzig E.M., Linder T., Wawrosch C., Uhrin P., Stuppner H. Discovery and resupply of pharmacologically active plant-derived natural products: a review // Biotechnol. Adv. 2015. V. 33. P. 1582. https://doi.org/10.1016/j.biotechadv.2015.08.001
Nakonechnaya O.V., Gafitskaya I.V., Grigorchuk V.P., Gorpenchenko T.Y., Bezdelev A.B., Zhuravlev Y.N. Polyphenol composition of Dracocephalum charkeviczii Prob. plants in in situ and in vitro conditions // Russ. J. Plant Physiol. 2022. V. 69:27. https://doi.org/10.1134/S1021443722010149
Kochkin D.V., Galishev B.A., Glagoleva E.S., Titova M.V., Nosov A.M. Rare triterpene glycoside of ginseng (ginsenoside malonyl-Rg 1) detected in plant cell suspension culture of Panax japonicus var. repens // Russ. J. Plant Physiol. 2017. V. 64. P. 649. https://doi.org/10.1134/S102144371705003X
Glagoleva E.S., Konstantinova S.V., Kochkin D.V., Ossipov V., Titova M.V., Popova E.V., Nosov A.M., Paek K.Y. 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 // Ind. Crops Prod. 2022. V. 177 P. 552114417 https://doi.org/10.1016/j.indcrop.2021.114417
Wu J., Zhong J.-J. Production of ginseng and its bioactive components in plant cell culture: Current technological and applied aspects // J. Biotechnol. 1999. V. 68. P. 89. https://doi.org/10.1016/s0168-1656(98)00195-3
Kochkin D.V., Glagoleva E.S., Galischev B.A., Spiridovich E.V., Nosov A.M., Reshetnikov V.N. Analysis of gynzenosides in the roots of Panax ginseng introduced in the central botanical garden of NAS of Belarus // Doklady of the National Academy of Sciences of Belarus. 2018. V. 62. P. 447. https://doi.org/10.29235/1561-8323-2018-62-4-447-454
Балеев Д.Н., Осипов В.И., Савин П.С., Байкова Ю.П., Сидельников Н.И. Сравнительный анализ состава и содержания гинзенозидов в каллусной культуре клеток и корне женьшеня обыкновенного, Panax ginseng // Биотехнология. 2022. Т. 38. № 2. С. 57. https://doi.org/10.56304/S0234275822020028
Song X., Wu H., Piao X., Yin Z., Yin Ch. Microbial transformation of ginsenosides extracted from Panax ginseng adventitious roots in an airlift bioreactor // Electron. J. Biotechnol. 2017. V. 26. P. 20. https://doi.org/10.1016/j.ejbt.2016.12.005
Журавлев Ю.Н., Гетманова Е.С., Музарок Т.И., Булгаков В.П. Способ микроразмножения женьшеня. Патент SU1824114A1, 1993.
Murashige T., Skoog F. A revised medium for rapid growth and bioassays with Tobacco tissue cultures // Physiol. Plant. 1962. V. 15. P. 473. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Жинкина Н.А., Воронова О.Н. К методике окраски эмбриологических препаратов // Ботанический журнал. 2000. Т. 85. С. 168.
Gorpenchenko T.Y., Kiselev K.V., Bulgakov V.P., Tchernoded G.K., Bragina E.A., Khodakovskaya M.V., Koren O.G., Batygina T.B., Zhuravlev Y.N. The Agrobacterium rhizogenes rolC-gene-induced somatic embryogenesis and shoot organogenesis in Panax ginseng transformed calluses // Planta. 2006. V. 223. P. 457. https://doi.org/10.1007/s00425-005-0102-2
Уварова Н.И., Маханьков В.В., Малиновская Г.В., Самошина Н.Ф., Атопкина Л.Н., Лихацкая Г.Н., Ким Н.Ю., Анисимов М.М., Еляков Г.Б. Химическая характеристика, сравнительное количественное определение и биологическая активность тритерпеновых гликозидов из дикого и плантационного женьшеня Panax ginseng C.A. Meyer, произрастающих в Приморском крае // Химико-фармацевтический журнал. 2000. Т. 34. С. 19.
Маханьков В.В., Бурундукова О.Л., Музарок Т.И., Уварова Н.И., Журавлев Ю.Н. Содержание гинзенозидов в листьях Panax ginseng C.A. Meyer в зависимости от возраста и условий произрастания // Растительные ресурсы. 2007. Т. 3. С. 107.
Chen Y., Zhao Z., Chen H., Yi T., Qin M., Liang Z. Chemical differentiation and quality evaluation of commercial asian and american ginsengs based on a UHPLC–QTOF/MS/MS metabolomics approach // Phytochem. Anal. 2015. V. 26 P. 145. https://doi.org/10.1002/pca.2546
Qiu S., Yang W.Z., Shi X.J., Yao C.L., Yang M., Liu X., Jiang B.-H., Wu W.-Y., Guo D.A. A green protocol for efficient discovery of novel natural compounds: characterization of new ginsenosides from the stems and leaves of Panax ginseng as a case study // Anal. Chim. Acta. 2015. V. 893 P. 65. https://doi.org/10.1016/j.aca.2015.08.048
Wang H.P., Zhang Y.B., Yang X.W., Zhao D.Q., Wang Y.P. Rapid characterization of ginsenosides in the roots and rhizomes of Panax ginseng by UPLC-DAD-QTOF-MS/MS and simultaneous determination of 19 ginsenosides by HPLC-ESI-MS // J. Ginseng Res. 2016. V. 40 P. 382. https://doi.org/10.1016/j.jgr.2015.12.001
Minh Duc N., Thoi Nham N., Kasai R., Ito A., Yamasaki K., Tanaka O. Saponins from Vietnamese ginseng, Panax vietnamensis HA et GRUSHV. Collected in Central Vietnam. I. // Chem. Pharm. Bull. 1993. V. 41. P. 2010. https://doi.org/10.1248/cpb.41.2010
Dictionary of food compounds additives, flavors, and ingredient / Ed. Sh. Yannai. 2004. A CRC ress Company Boca Raton, London, New York, Washington, D.C. 1782 p.
Van Le T.H., Lee G.J., Vu H.K.L., Kwon S.W., Nguyen N.K., Park J.H., Nguyen M.D. Ginseng saponins in different parts of Panax vietnamensis // Chem. Pharm. Bull. 2015. V. 63 P. 950. https://doi.org/10.1248/cpb.c15-00369
Peng M., Zhang T., Ding Y., Yi Y., Yang Y., Le J. Structure-based prediction of CAD response factors 1 of dammarane-type tetracyclic triterpenoid saponins and its application to the analysis of saponin contents in raw and processed Panax notoginseng // RSC Adv. 2016. V. 6. P. 36987. https://doi.org/10.1039/C6RA03193E
Wang H.-P., Wang Z.-J., Du J., Lin Z.-Z., Zhao C., Zhang R., Yin Q., Fan C.-L., Peng P., Wang Z.-B. Comprehensive identification of ginsenosides in the roots and rhizomes of Panax ginseng based on their molecular features-oriented precursor ions selection and targeted MS/MS analysis // Molecules. 2023. V. 28. P. 941. https://doi.org/10.3390/molecules28030941
Oh J.Y., Kim Y.J., Jang M.G., Joo S.C., Kwon W.S., Kim S.Y., Jung S.-K., Yang D.C. Investigation of ginsenosides in different tissues after elicitor treatment in Panax ginseng // J. Ginseng Res. 2014. V. 38. P. 270. https://doi.org/10.1016/j.jgr.2014.04.004
Rahimi S., Kim Y.J., Yang D.C. Production of ginseng saponins: elicitation strategy and signal transductions // Appl. Microbiol. Biotechnol. 2015. V. 99. P. 6987.
Chopra P., Chhillar H., Kim Y.J., Jo I.H., Kim S.T., Gupta R. Phytochemistry of ginsenosides: recent advancements and emerging roles // Crit. Rev. Food Sci. Nutr. 2023. V. 63. P. 613. https://doi.org/10.1080/10408398.2021.1952159
Moses T., Papadopoulou K.K., Osbourn A. Metabolic and functional diversity of saponins, biosynthetic intermediates and semi-synthetic derivatives // Crit. Rev. Biochem. Mol. Biol. 2014. V. 40. P. 439. https://doi.org/10.3109/10409238.2014.953628
Fehér A. Callus, dedifferentiation, totipotency, somatic embryogenesis: what these terms mean in the era of molecular plant biology? // Front. Plant Sci. 2019. V. 10: 536. https://doi.org/10.3389/fpls.2019.00536
Yang Y., Ju Z., Yang Y., Zhang Y., Yang L., Wang Z. Phytochemical analysis of Panax species: a review // J. Ginseng Res. 2021. V. 45. P. 1. https://doi.org/10.1016/j.jgr.2019.12.009
Yao C.L., Pan H.Q., Wang H., Yao S., Yang W.Z., Hou J.J., Jin Q.H., Wu W.Y., Guo D.A. Global profiling combined with predicted metabolites screening for discovery of natural compounds: characterization of ginsenosides in the leaves of Panax notoginseng as a case study // J. Chromatogr. 2018. V. 1538. P. 34. https://doi.org/10.1016/j.chroma.2018.01.040