Abstract
Euphorbia hebecarpa has been valued in traditional Iranian medicine; however, its carbohydrate composition has remained largely unexplored. The present study aimed to isolate, identify, and investigate selected biological properties of a carbohydrate fraction (EHC-1) from E. hebecarpa. EHC-1 was extracted using hot water extraction, followed by a two-step chromatographic procedure utilizing DEAE-52 cellulose and Sephadex G-200 columns. Partial structural characterization was performed using gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography- refractive index (HPLC-RID), Fourier transform infrared spectroscopy (FT-IR), and gas chromatography-flame ionization detection (GC-FID). In addition, X-ray diffraction (XRD) confirmed the semi-crystalline nature of the polysaccharide, and thermogravimetric analysis (TGA) was employed to evaluate its thermal stability. Compositional analyses identified EHC-1 as a polysaccharide with an average molecular weight of approximately 148 kDa. The relative monosaccharide composition comprised glucose (51.50%), fructose (35.70%), galactose (5.79%), arabinose (5.86%), and galacturonic acid (1.13%), as determined by the corrected peak area method incorporating relative response factors (RRFs). In vitro assays revealed that EHC-1 demonstrated concentration-dependent antioxidant activity in DPPH, ABTS, and hydroxyl radical scavenging assays, with relatively higher efficacy against hydroxyl radicals (IC50 = 2.43 ± 0.066 mg/mL). Furthermore, EHC-1 showed moderate antibacterial effects against both Escherichia coli and Staphylococcus aureus strains, with stronger effects against the latter.
Similar content being viewed by others
Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.
References
Podlogar, T. & Wallis, G. A. New horizons in carbohydrate research and application for endurance athletes. Sports Med. 52, 5–23 (2022).
K, R. et al. Exploring the diverse applications of carbohydrate macromolecules in food, pharmaceutical, and environmental technologies. Environ. Res. 240, 117521 (2024).
Digala, P. et al. Optimized extraction of sulfated polysaccharide from brown seaweed Sargassum polycystum and its evaluation for anti-cancer and wound healing potential. S. Afr. J. Bot. 151, 345–359 (2022).
Oyedepo, T. & Kayode, A. A. A. Bioactive carbohydrates, biological activities, and sources in functional foods and nutraceuticals. In Functional foods and nutraceuticals: bioactive components, formulations and innovations (eds Egbuna, C. & Dable-Tupas, G.) 39–74 (Springer, 2020).
Wu, Z.-W. et al. Structure–function insights of natural Ganoderma polysaccharides: Advances in biosynthesis and functional food applications. Nat. Prod. Bioprospect. 15, 1–33 (2025).
Zhang, W. et al. Immunomodulatory effects of Abelmoschus esculentus L. flower polysaccharide on spleen lymphocytes via TLR4-MAPK-NF-κB signaling pathways. South African J. Bot. 180, 520–530 (2025).
Abdel-Salam, E.A.-B., Nouh, M. I. & Elkholy, E. A. Analytical solution to the conformable fractional Lane-Emden type equations arising in astrophysics. Sci. Afr. 8, e00386 (2020).
Pahlevani, A. H., Liede-Schumann, S. & Akhani, H. Diversity, distribution, endemism and conservation status of Euphorbia (Euphorbiaceae) in SW Asia and adjacent countries. Plant Syst. Evol. 306, 1–26 (2020).
Stojilkovič, V., Záveská, E. & Frajman, B. From Western Asia to the Mediterranean basin: Diversification of the widespread Euphorbia nicaeensis alliance (Euphorbiaceae). Front. Plant Sci. 13, 1–24 (2022).
Salehi, B. et al. Euphorbia-derived natural products with potential for use in health maintenance. Biomolecules 9, 1–20 (2019).
Shi, Q.-W., Su, X.-H. & Kiyota, H. Chemical and pharmacological research of the plants in genus Euphorbia. Chem. Rev. 108, 4295–4327 (2008).
Sivaji, A., B, D. & A, M. S. Euphorbia hirta L.: A review on its ethnomedicinal uses, phytochemistry and pharmacology. World J. Pharm. Res. 3, 180–205 (2014).
Ghorbannia-Dellavar, S. et al. Antinociceptive activity of Iranian Euphorbia species in mice: Preliminary phytochemical analysis of Euphorbia malleata. S. Afr. J. Bot. 159, 532–543 (2023).
Amirghofran, Z. et al. Tumor cell death via apoptosis and improvement of activated lymphocyte cytokine secretion by extracts from Euphorbia hebecarpa and Euphorbia petiolata. Asian Pac. J. Cancer Prev. 20, 1979–1988 (2019).
Mohsenipour, Z. & Hassanshahian, M. Antibacterial activity of Euphorbia hebecarpa alcoholic extracts against six human pathogenic bacteria in planktonic and biofilm forms. Jundishapur J. Microbiol. 9, e34701 (2016).
Akhgar, M. R., Rajaei, P. & Aieen, S. Constituents of the essential oil of Euphorbia hebecarpa. Chem. Nat. Compd. 50, 929–930 (2014).
Liu, J., Sun, Y., Liu, L. & Yu, C. The extraction process optimization and physicochemical properties of polysaccharides from the roots of Euphorbia fischeriana. Int. J. Biol. Macromol. 49, 416–421 (2011).
Bao, L. et al. Structural characteristics and antioxidant activities of a novel polysaccharide from Euphorbia himalayensis root. Fitoterapia 176, 106009 (2024).
Xiang, N. et al. In vitro fecal fermentation of Euphorbia humifusa-derived polysaccharides and their protective effect against ulcerative colitis in mice. Foods 12, 751 (2023).
Athmouni, K., Belhaj, D., El Feki, A. & Ayadi, H. Optimization, antioxidant potential, modulatory effect and anti-apoptotic action in of Euphorbia bivonae polysaccharides on hydrogen peroxide-induced toxicity in human embryonic kidney cells HEK293. Int. J. Biol. Macromol. 116, 482–491 (2018).
Guo, H.-H. et al. Polysaccharide fraction from Triplostegia glandulifera Wall and its renoprotective effect in streptozotocin-induced diabetic mice by attenuating oxidative stress. Nat. Prod. Bioprospect. 14, 1–15 (2024).
Wang, Y. et al. Immunostimulatory effects mechanism of polysaccharide extracted from Acanthopanax senticosus on RAW 264.7 cells through activating the TLR/MAPK/NF-κB signaling pathway. Sci. Rep. 15, 13440 (2025).
Zhao, F. et al. Effect of Flammulina velutipes polysaccharide on mitochondrial apoptosis in lung adenocarcinomaA549 cells. Sci. Rep. 14, 16102 (2024).
Szymańska-Chargot, M., Pękala, P., Siemińska-Kuczer, A. & Zdunek, A. A determination of the composition and structure of the polysaccharides fractions isolated from apple cell wall based on FT-IR and FT-Raman spectra supported by PCA analysis. Food Hydrocoll. 150, 109688 (2024).
Mathlouthi, M. & Koenig, J. L. Vibrational spectra of carbohydrates. Adv. Carbohydr. Chem. Biochem. 44, 7–89 (1987).
Ahmadi, E., Rezadoost, H. & Moridi Farimani, M. Isolation, characterization, and antioxidant activity of neutral carbohydrates from Astragalus arbusculinus gum. S. Afr. J. Bot. 146, 669–675 (2022).
Saeidy, S. et al. Structural characterization and thermal behavior of a gum extracted from Ferula assa foetida L. Carbohydr. Polym. 181, 426–432 (2018).
Ballesteros, L. F., Cerqueira, M. A., Teixeira, J. A. & Mussatto, S. I. Characterization of polysaccharides extracted from spent coffee grounds by alkali pretreatment. Carbohydr. Polym. 127, 347–354 (2015).
Hou, Y., Ding, X. & Hou, W. Composition and antioxidant activity of water-soluble oligosaccharides from Hericium erinaceus. Mol. Med. Rep. 11, 3794–3799 (2015).
Fan, H., Meng, Q., Xiao, T. & Zhang, L. Partial characterization and antioxidant activities of polysaccharides sequentially extracted from Dendrobium officinale. J. Food Meas. Charact. 12, 1054–1064 (2018).
Zhao, Y. et al. Structural characterization, evaluation of lipid-lowering activity, and the structure-activity relationship of a pectic polysaccharide from the whole herb of Euphorbia humifusa. Carbohydr. Polym. 361, 123620 (2025).
Rezaei, A., Nasirpour, A. & Tavanai, H. Fractionation and some physicochemical properties of almond gum (Amygdalus communis L.) exudates. Food Hydrocoll. 60, 461–469 (2016).
Karimi, S., Ghanbarzadeh, B., Roufegarinejad, L. & Falcone, P. M. Polysaccharide extracted from Althaea officinalis L. root: New studies of structural, rheological and antioxidant properties. Carbohydr. Res. 510, 108438 (2021).
Wang, L. et al. Characterization, antioxidant and immunomodulatory effects of selenized polysaccharides from dandelion roots. Carbohydr. Polym. 260, 117796 (2021).
Wang, X. et al. Morphology and thermal characteristics of polysaccharide extracted by ultrasound from Ophiopogon japonicas. Am. J. Plant Sci. 09, 1699–1707 (2018).
Lo, T.C.-T., Chang, C. A., Chiu, K.-H., Tsay, P.-K. & Jen, J.-F. Correlation evaluation of antioxidant properties on the monosaccharide components and glycosyl linkages of polysaccharide with different measuring methods. Carbohydr. Polym. 86, 320–327 (2011).
Gao, D. et al. Structure characterization and antioxidant activity analysis of polysaccharides from Lanzhou Lily. Front. Nutr. 9, 1–13 (2022).
Bai, L. et al. Antioxidant activities of natural polysaccharides and their derivatives for biomedical and medicinal applications. Antioxidants 11, 2491 (2022).
Wang, T., Hu, S., Nie, S., Yu, Q. & Xie, M. Reviews on mechanisms of in vitro antioxidant activity of polysaccharides. Oxid. Med. Cell Long. 2016, 1–13 (2016).
Hentati, F. et al. Structural characterization and antioxidant activity of water-soluble polysaccharides from the Tunisian brown seaweed Cystoseira compressa. Carbohydr. Polym. 198, 589–600 (2018).
Ai-lati, A., Liu, S., Ji, Z., Zhang, H. & Mao, J. Structure and bioactivities of a polysaccharide isolated from Ganoderma lucidum in submerged fermentation. Bioeng. 8, 565–571 (2018).
Lu, Z. et al. Potential of antibacterial polysaccharides to serve as a promising alternative to conventional antibiotics: Challenges and future prospects. Int. J. Biol. Macromol. 322, 146960 (2025).
Palanisamy, S. et al. Antibacterial efficacy of a fucoidan fraction (Fu-F2) extracted from Sargassum polycystum. Int. J. Biol. Macromol. 15, 485–495 (2019).
Liu, J. et al. Alginate oligosaccharides: Production, biological activities, and potential applications. CRFSFS. 18, 1859–1881 (2019).
Sigríður Ólafsdóttir. Comparison of different assay methods and sample preparation procedures for the evaluation of the antibacterial activity of chitosan. https://skemman.is/bitstream/1946/35229/1/MSc%20thesis%202020%20Sigr%C3%AD%C3%B0ur%20%C3%93lafsd%C3%B3ttir.pdf (2020).
Ishak, A., Mazonakis, N., Spernovasilis, N., Akinosoglou, K. & Tsioutis, C. Bactericidal versus bacteriostatic antibacterials: Clinical significance, differences and synergistic potential in clinical practice. J. Antimicrob. Chemother. 80, 1–17 (2025).
Pankey, G. A. & Sabath, L. D. Clinical relevance of bacteriostatic versus bactericidal mechanisms of action in the treatment of gram-positive bacterial infections. Clin. Infect. Dis. 38, 864–870 (2004).
Ahmadi, E., Rezadoost, H., Alilou, M., Stuppner, H. & Moridi Farimani, M. Purification, structural characterization and antioxidant activity of a new arabinogalactan from Dorema ammoniacum gum. Int. J. Biol. Macromol. 194, 1019–1028 (2022).
DuBois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A. & Smith, F. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28, 350–356 (1956).
Chen, R.-Z. et al. Extraction, isolation, characterization and antioxidant activity of polysaccharides from Astragalus membranaceus. Ind. Crops Prod. 77, 434–443 (2015).
Wen, L. et al. Structural characterizations and α-glucosidase inhibitory activities of four Lepidium meyenii polysaccharides with different molecular weights. Nat. Prod. Bioprospect. 13, 1–9 (2023).
El-Messiry, H. M., Hamdan, A. M., Ghanem, N. B. & Hagar, M. Exopolysaccharide produced from Lactiplantibacillus plantarum HAN99 and its nanoparticle formulations in agricultural applications. Sci. Rep. 15, 19188 (2025).
Miller, G. L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31, 426–428 (1959).
Filisetti-Cozzi, T. M. & Carpita, N. C. Measurement of uronic acids without interference from neutral sugars. Anal. Biochem. 197, 157–162 (1991).
Hung, C. C. & Santschi, P. H. Spectrophotometric determination of total uronic acids in seawater using cation-exchange separation and pre-concentration by lyophilization. Anal. Chim. Acta 427, 111–117 (2001).
Walford, S. N. GC-MS as a tool for carbohydrate analysis in a research environment. Proc. Int. Soc. Sugar Cane Technol 27, 1–15 (2010).
Grob, R. L. & Barry, E. F. Qualitative and quantitative analysis by gas chromatography. In Modern practice of gas chromatography (eds Grob, R. L. & Barry, E. F.) 403–460 (Wiley, 2004).
Li, F. et al. Purification, structural characterization, antioxidant and emulsifying capabilities of exopolysaccharide produced by Rhodococcus qingshengii QDR4-2. J. Polym. Environ. 31, 64–80 (2023).
Chenga, H. et al. Structural characterization and antioxidant activities of polysaccharides extracted from Epimedium acuminatum. Carbohydr. Polym. 92, 63–68 (2013).
Yang, C. Optimation for preparation of oligosaccharides from flaxseed gum and evaluation of antioxidant and antitumor activities in vitro. Int. J. Biol. Macromol. 153, 1107–1116 (2020).
Guan, H. et al. Optimization of the process of acetylation and carboxymethylation for a polysaccharide from Gastrodia elata and antioxidant and immunomodulatory activities test. Sci. Rep. 15, 8460 (2025).
MehriArdestani, M. et al. Evaluation of antimicrobial activity of Trachyspermum ammi (L.) sprague essential oil and its active constituent, thymol, against vaginal pathogens. Tradit. Integr. Med. 5, 49–58 (2020).
Acknowledgments
Acknowledgments The authors acknowledge the support of Golestan University and Shahid Beheshti University.
Funding
No funding was received for conducting this study.
Author information
Authors and Affiliations
Contributions
Elham Ahmadi Jouibari: Methodology, Investigation, Formal analysis, Validation, Data curation, writing—original draft, Writing—review & editing. Mahdi Moridi Farimani: Supervision, Project administration, Visualization, Concetualization, Writing—review & editing. Mojtaba Asadollahi: Resources. All authors reviewed the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Ethical approval
Ethics approval was not required for this research.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Juybari, E.A., Farimani, M.M. & Asadollahi, M. Partial characterization and evaluation of antioxidant and antibacterial activities of a water soluble polysaccharide isolated from Euphorbia hebecarpa roots. Sci Rep (2026). https://doi.org/10.1038/s41598-026-42880-7
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-42880-7
