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ORIGINAL ARTICLE
Cultivar- and Growth-Stage-Dependent Variability of Saponins in Roots and Leaves of Beta vulgaris L. Characterized by Liquid Chromatography Coupled with Mass Spectrometry
 
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1
Department of Chemical Technology and Environmental Analysis, Faculty of Chemical Engineering and Technology, Cracow University of Technology, Warszawska 24, 31-155 Kraków, Poland
 
2
Department of Agriculture and Forest Sciences (DAFNE), University of Tuscia, Via San Camillo de Lellis, 01100 Viterbo, Italy
 
These authors had equal contribution to this work
 
 
Submission date: 2025-11-24
 
 
Acceptance date: 2026-03-02
 
 
Online publication date: 2026-03-20
 
 
Publication date: 2026-03-20
 
 
Corresponding author
Aneta Spórna-Kucab   

Department of Chemical Technology and Environmental Analysis, Cracow University of Technology, Warszawska 24, 31-155, Kraków, Poland
 
 
Pol. J. Food Nutr. Sci. 2026;76(1):106-117
 
KEYWORDS
TOPICS
ABSTRACT
Beta vulgaris L. is an important food crop and a rich source of bioactive triterpene saponins. This study evaluated cultivar- and growth-stage-dependent variability of saponins in leaves and roots of three beet cultivars (Round Dark Red, Cylindra, and Snow Ball) and in leaves of Swiss chard (Rhubarb Chard), harvested at seven harvest dates between June and September 2024. Saponins were identified and quantified using liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS). A total of 32 triterpene saponins representing oleanane-type, akebonoic acid-, hederagenin-, and gypsogenin-derived aglycones were detected. Pronounced organ-dependent differences were observed. Total saponin content ranged from 386 to 10,414 mg/kg fresh weight (FW) in leaves and from 1,170 to 23,298 mg/kg FW in roots. Two-way analysis of variance confirmed highly significant effects of cultivar, harvest time, and their interaction on total saponin levels in both leaves and roots (all p<0.0001). Roots exhibited a broader content range and a pronounced mid-season maximum, whereas leaf saponin levels generally peaked in the mid-to-late season in a genotype-dependent pattern. Major saponins (Act-UrA-akebonoic acid and betavulgarosides II, III, IV, and VII) predominated in the quantitative profile and exhibited coordinated seasonal variation. Multivariate analyses (principal component analysis and hierarchical cluster analysis) clearly separated samples according to plant organ and further resolved cultivar- and season-related patterns. Overall, saponin accumulation in B. vulgaris is strongly regulated by organ type, genotype, and growth stage, emphasizing the importance of cultivar selection and harvest timing for maximizing bioactive potential.
FUNDING
This research was funded in whole by National Science Centre, Poland for the years 2024–2027 (Project No. UMO-2023/49/N/NZ9/01706).
CONFLICT OF INTEREST
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
REFERENCES (48)
1.
Arslan I. (2017). A new acylated and oleanane-type triterpenoid saponin from Gypsophila arrostii roots. International Journal of Food Properties, 20(3), 507–513. https://doi.org/10.1080/10942912.2016.1168437
 
2.
Arslan I. (2020). Quillaic acid–containing saponin-based immunoadjuvants trigger early immune responses. Revista Brasileira de Farmacognosia, 30(4), 467–473. https://doi.org/10.1007/s43450-020-00080-y
 
3.
Arslan I., Cenzano A.M. (2021). Characterization of triterpene saponins from Gypsophila arrostii by electrospray ionization ion trap multiple-stage mass spectrometry. Revista Brasileira de Farmacognosia, 31(1), 107–111. https://doi.org/10.1007/s43450-021-00125-w
 
4.
Biswas T., Dwivedi U.N. (2019). Plant triterpenoid saponins: biosynthesis, in vitro production, and pharmacological relevance. Protoplasma, 256, 1463–1486. https://doi.org/10.1007/s00709-019-01411-0
 
5.
Böttger S., Melzig M.F. (2013). The influence of saponins on cell membrane cholesterol. Bioorganic & Medicinal Chemistry, 21(22), 7118–7124. https://doi.org/10.1016/j.bmc.2013.09.008
 
6.
Castellano J.M., Ramos-Romero S., Perona J.S. (2022). Oleanolic acid: Extraction, Characterization and biological activity. Nutrients, 14(3), art. no. 623. https://doi.org/10.3390/nu14030623
 
7.
Cheng L., Shi L., Wu J., Zhou X., Li X., Sun X., Zhu L., Xia T.S., Ding Q. (2018). A hederagenin saponin isolated from Clematis ganpiniana induces apoptosis in breast cancer cells via the mitochondrial pathway. Oncology Letters, 15(2), 1737–1743. https://doi.org/10.3892/ol.2017.7494
 
8.
Ciftci H.I., Ozturk S.E., Ali T.F.S., Radwan M.O., Tateishi H., Koga R., Ocak Z., Can M., Otsuka M., Fujita M. (2018). The first pentacyclic triterpenoid gypsogenin derivative exhibiting anti-ABL1 kinase and anti-chronic myelogenous leukemia activities. Biological and Pharmaceutical Bulletin, 41(4), 570–574. https://doi.org/10.1248/bpb.b17-00902
 
9.
Dirir A.M., Daou M., Yousef A.F., Yousef L.F. (2022). A review of alpha-glucosidase inhibitors from plants as potential candidates for the treatment of type-2 diabetes. Phytochemistry Reviews, 21, 1049–1079. https://doi.org/10.1007/s11101-021-09773-1
 
10.
Edelmann M., Dawid C., Hochreiter K., Ralla T., Stark T.D., Salminen H., Weiss J., Hofmann T. (2020a). Molecularization of foam-active saponins from sugar beet side streams (Beta vulgaris ssp. vulgaris var. altissima). Journal of Agricultural and Food Chemistry, 68(39), 10962–10974. https://doi.org/10.1021/acs.jafc.0c04603
 
11.
Edelmann M., Dawid C., Ralla T., Stark T.D., Salminen H., Weiss J., Hofmann T. (2020b). Fast and sensitive LC–MS/MS method for the quantitation of saponins in various sugar beet materials. Journal of Agricultural and Food Chemistry, 68(50), 15027–15035. https://doi.org/10.1021/acs.jafc.0c05836
 
12.
Engholm D.H., Kilian M., Goodsell D.S., Andersen E.S., Kjærgaard R.S. (2017). A visual review of the human pathogen Streptococcus pneumoniae. FEMS Microbiology Reviews, 41(6), 854–879. https://doi.org/10.1093/femsre/fux037
 
13.
Favel A., Steinmetz M.D., Regli P., Vidal-Ollivier E., Elias R., Balansard G. (1994). In vitro antifungal activity of triterpenoid saponins. Planta Medica, 60(1), 50–53. https://doi.org/10.1055/s-2006-959407
 
14.
Francis G., Kerem Z., Makkar H.P., Becker K. (2002). The biological action of saponins in animal systems: a review. British Journal of Nutrition, 88(6), 587–605. https://doi.org/10.1079/BJN2002725
 
15.
Günther A., Bednarczyk-Cwynar B. (2025). Oleanolic acid: A promising antioxidant – sources, mechanisms of action, therapeutic potential, and enhancement of bioactivity. Antioxidants, 14(5), art. no. 598. https://doi.org/10.3390/antiox14050598
 
16.
Gupta N. (2022). A review on recent developments in the anticancer potential of oleanolic acid and its analogs (2017-2020). Mini-Reviews in Medicinal Chemistry, 22(4), 600–616. https://doi.org/10.2174/1389557521666210810153627
 
17.
Ida Y., Satoh Y., Katsumata M., Nagasao M., Shoji J. (1995). Achyranthosides C and D, novel glucuronide saponins from Achyranthes fauriei root. Chemical and Pharmaceutical Bulletin, 43(5), 896–898. https://doi.org/10.1248/cpb.43.896
 
18.
Jolly A., Hour Y., Lee Y.C. (2024). An outlook on the versatility of plant saponins: A review. Fitoterapia, 174, art. no. 105858. https://doi.org/10.1016/j.fitote.2024.105858
 
19.
Kaur R., Mishra V., Gupta S., Sharma S., Vaishnav A., Singh S.V. (2024). Industrial and environmental applications of plant-derived saponins: An overview and future prospective. Journal of Plant Growth Regulation, 43, 3012–3026. https://doi.org/10.1007/s00344-023-11201-x
 
20.
Kinjo J., Okawa M., Udayama M., Shono Y., Hirakawa T., Shii Y., Nohara T. (1999). Hepatoprotective and hepatotoxic actions of oleanolic acid-type triterpenoidal glucuronides on rat primary hepatocyte cultures. Chemical and Pharmaceutical Bulletin, 47(2), 290–292. https://doi.org/10.1248/cpb.47.290
 
21.
Krasteva I., Yotova M., Yosifov D., Benbassat N., Jenett-Siems K., Konstantinov S. (2014). Cytotoxicity of gypsogenic acid isolated from Gypsophila trichotoma. Pharmacognosy Magazine, 10(Sup. 2), S430–S433.
 
22.
Kregiel D., Berlowska J., Witonska I., Antolak H., Proestos C., Babic M., Babic L., Zhang B. (2017). Chapter 6 – Saponin-based, biological-active surfactants from plants. In R. Najjar (Ed.), Application and Characterization of Surfactants, IntechOpen, pp. 184–197. https://doi.org/10.5772/68062
 
23.
Lisiak N.M., Lewicka I., Kaczmarek M., Kujawski J., Bednarczyk-Cwynar B., Zaprutko L., Rubis B. (2021). Oleanolic acid’s semisynthetic derivatives HIMOXOL and br-HIMOLID show proautophagic potential and inhibit migration of HER2-positive breast cancer cells in vitro. International Journal of Molecular Sciences, 22(20), art. no. 11273. https://doi.org/10.3390/ijms222011273
 
24.
Liu J., Lu Y.F., Wu Q., Xu S.F., Shi F.G., Klaassen C.D. (2019). Oleanolic acid reprograms the liver to protect against hepatotoxicants, but is hepatotoxic at high doses. Liver International, 39(3), 427–439. https://doi.org/10.1111/liv.13940
 
25.
Mikołajczyk-Bator K., Błaszczyk A., Czyżniejewski M., Kachlicki P. (2016a). Characterisation and identification of triterpene saponins in the roots of red beets (Beta vulgaris L.) using two HPLC–MS systems. Food Chemistry, 192, 979–990. https://doi.org/10.1016/j.foodchem.2015.07.111
 
26.
Mikołajczyk-Bator K., Błaszczyk A., Czyżniejewski M., Kachlicki P. (2016b) Identification of saponins from sugar beet (Beta vulgaris) by low and high-resolution HPLC–MS/MS. Journal of Chromatography B, 1029–1030, 36–47. https://doi.org/10.1016/j.jchromb.2016.06.038
 
27.
Mikołajczyk-Bator K., Błaszczyk A., Czyżniejewski M., Kachlicki P., Stochmal A. (2024). Metabolite profiling of triterpene saponins from different red beetroot cultivars using ultra-high performance liquid chromatography high-resolution mass spectrometry. Journal of Food Composition and Analysis, 130, art. no. 106141. https://doi.org/10.1016/j.jfca.2024.106141
 
28.
Mroczek A., Klimczak U., Kowalczyk M. (2021). Determination of saponins in leaves of four Swiss chard (Beta vulgaris L.) cultivars by UHPLC-CAD/QTOF-MS/MS. Polish Journal of Food and Nutrition Sciences, 71(2), 147–159. https://doi.org/10.31883/pjfns/134623
 
29.
Mroczek A. (2015). Phytochemistry and bioactivity of triterpene saponins from Amaranthaceae family. Phytochemistry Reviews, 14, 577–605. https://doi.org/10.1007/s11101-015-9394-4
 
30.
Mroczek A., Kapusta I., Janda B., Janiszowska W. (2012). Triterpene saponin content in the roots of red beet (Beta vulgaris L.) cultivars. Journal of Agricultural and Food Chemistry, 60(50), 12397–12402. https://doi.org/10.1021/jf303952x
 
31.
Mroczek A., Kapusta I., Stochmal A., Janiszowska W. (2019). MS/MS and UPLC-MS profiling of triterpenoid saponins from leaves and roots of four red beet (Beta vulgaris L.) cultivars. Phytochemistry Letters, 30, 333–337. https://doi.org/10.1016/j.phytol.2019.02.015
 
32.
Sen A. (2020). Prophylactic and therapeutic roles of oleanolic acid and its derivatives in several diseases. World Journal of Clinical Cases, 8(10), 1767–1792. https://doi.org/10.12998/wjcc.v8.i10.1767
 
33.
Sharma K., Sharma S., Vaishnav A., Jain R., Singh D., Singh H.B., Goel A., Singh S. (2022). Salt-tolerant PGPR strain Priestia endophytica SK1 promotes fenugreek growth under salt stress by inducing nitrogen assimilation and secondary metabolites. Journal of Applied Microbiology, 133(5), 2802–2813. https://doi.org/10.1111/jam.15735
 
34.
Spórna-Kucab A., Tekieli A., Skalicka-Woźniak K., Grzegorczyk A., Świergosz T., Wybraniec S. (2022). Characterization of triterpene saponin composition of white, yellow and red beetroot (Beta vulgaris L.). Polish Journal of Food and Nutrition Sciences, 72(2), 159–170. https://doi.org/10.31883/pjfns/149515
 
35.
Spórna-Kucab A.E., Wybraniec S. (2020). High-speed counter-current chromatography in separation and identification of saponins from Beta vulgaris L. cultivar Red Sphere. Polish Journal of Food and Nutrition Sciences, 70(1), 67–74. https://doi.org/10.31883/pjfns/116416
 
36.
Su F., Sui X., Xu J., Liu Q., Li J., Liu W., Xu L., Zhang Z., Tao F. (2024). Hederagenin suppresses ovarian cancer via targeting mitochondrial fission through dynamin-related protein 1. European Journal of Pharmacology, 963, art. no. 176188. https://doi.org/10.1016/j.ejphar.2023.176188
 
37.
Sun K.P., Zhao T.T., Liu L., Mu X.D., Sun J.Y. (2023). Anticancer structure-activity relationships and potential target exploration of the natural product gypsogenin. ChemistrySelect, 8(10), art. no. e202300072. https://doi.org/10.1002/slct.202300072
 
38.
De Tommasi N., Conti C., Stein M.L., Pizza C. (1991). Structure and in vitro antiviral activity of triterpenoid saponins from Calendula arvensis. Planta Medica, 57(3), 250–253. https://doi.org/10.1055/s-2006-960084
 
39.
Tekieli A., Kowalczyk M., Wybraniec S., Spórna-Kucab A. (2026). pH-dependent separation and identification of saponins from Beta vulgaris L. using high-speed countercurrent chromatography and high-resolution mass spectrometry. Journal of Chromatography A, 1769, art. no. 466643. https://doi.org/10.1016/j.chroma.2025.466643
 
40.
Vincken J.P., Heng L., de Groot A., Gruppen H. (2007). Saponins, classification and occurrence in the plant kingdom. Phytochemistry, 68(3), 275–297. https://doi.org/10.1016/j.phytochem.2006.10.008
 
41.
Wang J., Xu Q.L., Zheng M.F., Ren H., Lei T., Wu P., Zhou Z.Y., Wei X.Y., Tan J.W. (2014). Bioactive 30-noroleanane triterpenes from the pericarps of Akebia trifoliata. Molecules, 19(4), 4301–4312. https://doi.org/10.3390/molecules19044301
 
42.
Wen F., Chen S., Wang Y., Wu Q., Yan J., Pei J., Zhou T. (2023). The synthesis of Paris saponin VII mainly occurs in leaves and is promoted by light intensity. Frontiers in Plant Science, 14, art. no. 1199215. https://doi.org/10.3389/fpls.2023.1199215
 
43.
Xie W., Fang X., Li H., Lu X., Yang D., Han S., Bi Y. (2023). Advances in the anti-tumor potential of hederagenin and its analogs. European Journal of Pharmacology, 959, art. no. 176073. https://doi.org/10.1016/j.ejphar.2023.176073
 
44.
Yoon Y., Choi K. H. (2010). Identification of inhibitory effect on Streptococcus mutans by oleanolic acid. Journal of Life Science, 20(3), 321–325. https://doi.org/10.5352/JLS.2010.20.3.321
 
45.
Yoshikawa M., Murakami T., Kadoya M., Matsuda H., Muraoka O., Yamahara J., Murakami N. (1996). Medicinal foodstuffs. III. Sugar beet. (1): Hypoglycemic oleanolic acid oligoglycosides, betavulgarosides, I, II, III, and IV, from the root of Beta vulgaris L. (Chenopodiaceae). Chemical and Pharmaceutical Bulletin, 44(6), 1212–1217. https://doi.org/10.1248/cpb.44.1212
 
46.
Yoshikawa M., Murakami T., Kadoya M., Yamahara J., Matsuda H. (1998). Medicinal foodstuffs. XV. Sugar beet. (2): Structures of betavulgarosides V, VI, VII, VIII, IX and X from the roots and leaves of sugar beet (Beta vulgaris L., Chenopodiaceae). Chemical and Pharmaceutical Bulletin, 46(11), 1758–1763. https://doi.org/10.1248/cpb.46.1758
 
47.
Yu X.L., He Y. (2018). Tea saponins: effective natural surfactants beneficial for soil remediation, from preparation to application. RSC Advances, 8(43), 24312–24321. https://doi.org/10.1039/C8RA02859A
 
48.
Zeng J., Huang T., Xue M., Chen J., Feng L., Du R., Feng Y. (2018). Current knowledge and development of hederagenin as a promising medicinal agent: a comprehensive review. RSC Advances, 8(43), 24188–24202. https://doi.org/10.1039/C8RA03666G
 
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