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Ultrasound-assisted aqueous two-phase extraction of flavonoids from erigeron breviscapus: process optimization, structural characterization, antioxidant study, and DFT calculation
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  • Published: 03 March 2026

Ultrasound-assisted aqueous two-phase extraction of flavonoids from erigeron breviscapus: process optimization, structural characterization, antioxidant study, and DFT calculation

  • Huiqin Qian1,
  • Menglin Wang1,
  • Haibo Xu1,
  • Kun Feng1,
  • Yaxuan Li1,
  • Yingfan Hu1 &
  • …
  • Yanling Li1 

Scientific Reports , Article number:  (2026) Cite this article

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We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Biochemistry
  • Chemistry
  • Drug discovery
  • Plant sciences

Abstract

The well-known medicinal plant Erigeron breviscapus has long been used to treat cerebral embolism, cerebral thrombosis, and cerebral hemorrhage. Response surface methodology (RSM) was applied to optimize the ultrasonic-assisted extraction process of total flavonoids from Erigeron breviscapus (EBTF) using aqueous two-phase system. The flavonoids from E. breviscapus were qualitatively identified using UPLC-Q-TOF-MS/MS. The capacity of EBTF to scavenge ·OH was used to assess its antioxidant activity. To determine the active sites in the primary bioactive components that scavenge ·OH, density functional theory (DFT) calculations were conducted. Total flavonoid content (TFC) from E. breviscapus was 48.53 mg/g under ideal conditions with PEG2000 mass fraction of 16%, (NH4)2SO4 mass fraction of 14%, ultrasound time of 41 min, and liquid-solid ratio of 35 mL/g. 28 flavonoids have been tentatively identified in E. breviscapus via ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS). Furthermore, EBTF demonstrated moderate hydroxyl radical scavenging capacity, with scavenging rate of 60.68% at 3.9 mg/mL. The 6-OH site of scutellarin was the core active site for scavenging hydroxyl radicals. The findings provide both theoretical and experimental support for the in-depth development of EBTF as a natural antioxidant.

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Data availability

Data is available from the corresponding author on reasonable request.

References

  1. Zhu, Z. et al. Innovative development path of ethnomedicines: a case study. Front. Med. 11, 297–305. https://doi.org/10.1007/s11684-017-0513-z (2017).

    Google Scholar 

  2. Wu, R. et al. Advances in Chemical Constituents, Clinical Applications, Pharmacology, Pharmacokinetics and Toxicology of Erigeron breviscapus. Front. Pharmacol. 12, 656335. https://doi.org/10.3389/fphar.2021.656335 (2021).

    Google Scholar 

  3. Li, Y. et al. Development of an efficient extraction and enrichment method for total flavonoids compounds from Erigeron breviscapus using ultrasound-assisted extraction and macroporous resin adsorption. Prep. Biochem. Biotechnol. 55, 1245–1256. https://doi.org/10.1080/10826068.2025.2502757 (2025).

    Google Scholar 

  4. Zhang, Y. et al. Fermented Erigeron breviscapus flavonoids: anti-pseudorabies virus efficacy and mechanisms in vitro and in vivo. Front. veterinary Sci. 12, 1562879. https://doi.org/10.3389/fvets.2025.1562879 (2025).

    Google Scholar 

  5. Zhu, J. T. et al. Estrogenic and neuroprotective properties of scutellarin from Erigeron breviscapus: a drug against postmenopausal symptoms and Alzheimer’s disease. Planta Med. 75, 1489–1493. https://doi.org/10.1055/s-0029-1185776 (2009).

    Google Scholar 

  6. Tao, Y. H., Jiang, D. Y., Xu, H. B. & Yang, X. L. Inhibitory effect of Erigeron breviscapus extract and its flavonoid components on GABA shunt enzymes. Phytomedicine: Int. J. phytotherapy phytopharmacology. 15, 92–97. https://doi.org/10.1016/j.phymed.2007.06.009 (2008).

    Google Scholar 

  7. Chledzik, S., Strawa, J., Matuszek, K. & Nazaruk, J. Pharmacological Effects of Scutellarin, An Active Component of Genus Scutellaria and Erigeron: A Systematic Review. Am. J. Chin. Med. 46, 319–337. https://doi.org/10.1142/s0192415x18500167 (2018).

    Google Scholar 

  8. Fan, H. et al. Metabolism and Pharmacological Mechanisms of Active Ingredients in Erigeron breviscapus. Curr. Drug Metab. 22, 24–39. https://doi.org/10.2174/1389200221666201217093255 (2021).

    Google Scholar 

  9. Zhang, X., Han, M., Han, S. & Zong, W. Aqueous two-phase system (ATPS): from basic science to applications. RSC Adv. 15, 9041–9054. https://doi.org/10.1039/d4ra08232j (2025).

    Google Scholar 

  10. Lin, Y. et al. Content determination of the flavonoids in the different parts and different species of Abelmoschus esculentus L. by reversed phase-high performance liquid chromatograph and colorimetric method. Pharmacognosy magazine. 10, 278–284. https://doi.org/10.4103/0973-1296.137368 (2014).

    Google Scholar 

  11. Huang, H. et al. Study on the extraction, antioxidant and prebiotic activity of the polysaccharides from the fruits of Phyllanthus emblica L. Front. Nutr. 12, 1607077. https://doi.org/10.3389/fnut.2025.1607077 (2025).

    Google Scholar 

  12. Yang, H. et al. Scutellarin inhibits ferroptosis by promoting cellular antioxidant capacity through regulating Nrf2 signaling. Acta Biochim. Biophys. Sin. https://doi.org/10.3724/abbs.2025112 (2025).

    Google Scholar 

  13. Gao, D. et al. Extraction, structural characterization, and antioxidant activity of polysaccharides derived from Arctium lappa L. Front. Nutr. 10 https://doi.org/10.3389/fnut.2023.1149137 (2023).

  14. Chong, K. Y., Stefanova, R., Zhang, J. & Brooks, M. S. L. Aqueous two-phase extraction of bioactive compounds from haskap leaves (Lonicera caerulea): Comparison of salt/ethanol and sugar/propanol systems. Sep. Purif. Technol. 252, 117399. https://doi.org/10.1016/j.seppur.2020.117399 (2020).

    Google Scholar 

  15. Wang, Q., Jia, C., Chang, L. & Tang, H. Optimization of Aqueous Two-phase Extraction of Total Flavonoids from Actinidia kolomikta Leaves. Food Sci. 32, 167–171 (2011).

    Google Scholar 

  16. Lai, J. et al. Ultrasound-assisted deep eutectic solvent extraction of flavonol glycosides from Ginkgo biloba: Optimization of efficiency and mechanism. Ultrason. Sonochem. 114, 107254. https://doi.org/10.1016/j.ultsonch.2025.107254 (2025).

    Google Scholar 

  17. Wu, Z. et al. Extraction of American ginseng polysaccharide by ultrasound-assisted deep eutectic solvents-based three-phase partitioning: Process optimization, structural characterization, and anti-ulcerative colitis study. Ultrason. Sonochem. 112, 107206. https://doi.org/10.1016/j.ultsonch.2024.107206 (2025).

    Google Scholar 

  18. Qian, Y. et al. Optimization of Polysaccharides Extraction from Seed Melon by Response Surface Methodology and Its Hypolipidemic Effects in Vitro. Sci. Technol. Food Ind. 41, 101–107. https://doi.org/10.13386/j.issn1002-0306.2020.02.017 (2020).

    Google Scholar 

  19. Pan, L. et al. Enzymatic Pretreatment Solvent-free Microwave Extraction of Fresh Cirsium setosum Essential Oil and Analysis of Antioxidant Activity. Sci. Technol. Food Ind. 46, 132–145. https://doi.org/10.13386/j.issn1002-0306.2024100100 (2025).

    Google Scholar 

  20. Silva, D. B. et al. Mass spectrometry of flavonoid vicenin-2, based sunlight barriers in Lychnophora species. Sci. Rep. 4, 4309. https://doi.org/10.1038/srep04309 (2014).

    Google Scholar 

  21. Yang, Y. et al. Quantitative and Qualitative Analysis of Flavonoids and Phenolic Acids in Snow Chrysanthemum (Coreopsis tinctoria Nutt.) by HPLC-DAD and UPLC-ESI-QTOF-MS. Molecules 21 (2016).

  22. El-Banna, A. A. et al. Metabolic profiling of Lantana camara L. using UPLC-MS/MS and revealing its inflammation-related targets using network pharmacology-based and molecular docking analyses. Sci. Rep. 12, 14828. https://doi.org/10.1038/s41598-022-19137-0 (2022).

    Google Scholar 

  23. Tanaka, H., Tanaka, T., Hosoya, A., Kitade, Y. & Etoh, H. Three isoflavanones from Erythrina orientalis. Phytochemistry 48, 355–357. https://doi.org/10.1016/S0031-9422(97)01012-1 (1998).

    Google Scholar 

  24. Lage, G. A. et al. The first report on flavonoid isolation from Annona crassiflora Mart. Nat. Prod. Res. 28, 808–811. https://doi.org/10.1080/14786419.2014.885518 (2014).

    Google Scholar 

  25. Guan, Z. et al. Identification and quantitation of phenolic compounds from the seed and pomace of Perilla frutescens using HPLC/PDA and HPLC-ESI/QTOF/MS/MS. Phytochemical analysis : PCA 25, 508–513, doi:10.1002/pca.2521 (2014)

  26. Liu, Y. et al. Exploring the mechanisms of Huangqin Qingfei Decoction on acute lung injury by LC-MS combined network pharmacology analysis. Phytomedicine: Int. J. phytotherapy phytopharmacology. 134, 155979. https://doi.org/10.1016/j.phymed.2024.155979 (2024).

    Google Scholar 

  27. Li, S., Wang, R., Hu, X., Li, C. & Wang, L. Bio-affinity ultra-filtration combined with HPLC-ESI-qTOF-MS/MS for screening potential α-glucosidase inhibitors from Cerasus humilis (Bge.) Sok. leaf-tea and in silico analysis. Food Chem. 373, 131528. https://doi.org/10.1016/j.foodchem.2021.131528 (2022).

    Google Scholar 

  28. Wang, J. et al. LC-MS/MS-based chemical profiling of water extracts of Moringa oleifera leaves and pharmacokinetics of their major constituents in rat plasma. Food chemistry: X. 23, 101585. https://doi.org/10.1016/j.fochx.2024.101585 (2024).

    Google Scholar 

  29. Meng, C. et al. Metabolic profiling comparison of isovitexin in normal and kidney stone model rats by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. J. Sep. Sci. 43, 2363–2379. https://doi.org/10.1002/jssc.201901169 (2020).

    Google Scholar 

  30. Li, F. et al. Screening of free radical scavengers from Erigeron breviscapus using on-line HPLC-ABTS/DPPH based assay and mass spectrometer detection. Free Radic. Res. 46, 286–294. https://doi.org/10.3109/10715762.2011.652628 (2012).

    Google Scholar 

  31. Zhang, Z. et al. Characterization and quantification of the chemical compositions of Scutellariae Barbatae herba and differentiation from its substitute by combining UHPLC-PDA-QTOF-MS/MS with UHPLC-MS/MS. J. Pharm. Biomed. Anal. 109, 62–66. https://doi.org/10.1016/j.jpba.2015.02.025 (2015).

    Google Scholar 

  32. Fernandes, J. M. et al. Inhibitory Effects of Hydroethanolic Leaf Extracts of Kalanchoe brasiliensis and Kalanchoe pinnata (Crassulaceae) against Local Effects Induced by Bothrops jararaca Snake Venom. PloS one. 11, e0168658. https://doi.org/10.1371/journal.pone.0168658 (2016).

    Google Scholar 

  33. Alhumaydhi, F. A., Aljohani, A. S. M. & Elsharkawy, E. R. UPLC/ESI-MS Phytochemical Screening of Deverra tortuosa Haematological and Histopathological Studies and Streptozotocin-Induced Diabetes in Rat. Evidence-based Complement. Altern. medicine: eCAM. 2021 (4718854). https://doi.org/10.1155/2021/4718854 (2021).

  34. Lin, S., Simal-Gandara, J., Cao, H. & Xiao, J. The stability and degradation products of polyhydroxy flavonols in boiling water. Curr. Res. food Sci. 6, 100509. https://doi.org/10.1016/j.crfs.2023.100509 (2023).

    Google Scholar 

  35. Yin, N. W. et al. Identification and Characterization of Major Constituents in Different-Colored Rapeseed Petals by UPLC-HESI-MS/MS. J. Agric. Food Chem. 67, 11053–11065. https://doi.org/10.1021/acs.jafc.9b05046 (2019).

    Google Scholar 

  36. Zhao, L. et al. UPLC-Q-Exactive/MS based analysis explore the correlation between components variations and anti-influenza virus effect of four quantified extracts of Chaihu Guizhi decoction. J. Ethnopharmacol. 319, 117318. https://doi.org/10.1016/j.jep.2023.117318 (2024).

    Google Scholar 

  37. Jacobsen, S. S., Knob, F. C., Simon, A. P. & Oldoni, T. L. C. Selective Extraction Process and Characterization of Antioxidant Phenolic Compounds from Pereskia aculeata Leaves Using UPLC-ESI-Q-TOF-MS/MS. ACS omega. 9, 37374–37385. https://doi.org/10.1021/acsomega.4c05652 (2024).

    Google Scholar 

  38. Zhao, Y. et al. Effect of drying processes on prenylflavonoid content and antioxidant activity of Epimedium koreanum Nakai. J. food drug Anal. 26, 796–806. https://doi.org/10.1016/j.jfda.2017.05.011 (2018).

    Google Scholar 

  39. Liu, H. et al. Prostate Distribution and Metabolic Characteristics of Four Representative Flavones after Oral Administration of the Aerial Part of Glycyrrhiza uralensis in Rats. Molecules 27 https://doi.org/10.3390/molecules27103245 (2022). Pharmacokinetics.

  40. Kumari, S., Elancheran, R., Kotoky, J. & Devi, R. Rapid screening and identification of phenolic antioxidants in Hydrocotyle sibthorpioides Lam. by UPLC-ESI-MS/MS. Food Chem. 203, 521–529. https://doi.org/10.1016/j.foodchem.2016.02.101 (2016).

    Google Scholar 

  41. Meng, X. et al. Metabolism of eriocitrin in the gut and its regulation on gut microbiota in mice. Front. Microbiol. 13 https://doi.org/10.3389/fmicb.2022.1111200 (2022).

  42. Gan, X. et al. Identification of Xanthine Oxidase Inhibitors from Celery Seeds Using Affinity Ultrafiltration-Liquid Chromatography-Mass Spectrometry. Molecules 28 https://doi.org/10.3390/molecules28166048 (2023).

  43. Zhao, X. et al. Analysis of the permeable and retainable components of Cayratia japonica ointment through intact or broken skin after topical application by UPLC-Q-TOF-MS/MS combined with in vitro transdermal assay. J. Pharm. Biomed. Anal. 238, 115853. https://doi.org/10.1016/j.jpba.2023.115853 (2024).

    Google Scholar 

  44. Fu, X. et al. UPLC-Triple-TOF/MS characterization of phenolic constituents and the influence of natural deep eutectic solvents on extraction of Carya cathayensis Sarg. peels: Composition, extraction mechanism and in vitro biological activities. Food Chem. 370, 131042. https://doi.org/10.1016/j.foodchem.2021.131042 (2022).

    Google Scholar 

  45. Zhang, J. et al. Metabolite Identification and Pharmacokinetic Profiling of Isoflavones from Black Soybean in Rats Using Ultrahigh-Performance Liquid Chromatography with Linear-Ion-Trap-Orbitrap and Triple-Quadrupole Tandem Mass Spectrometry. J. Agric. Food Chem. 66, 12941–12952. https://doi.org/10.1021/acs.jafc.8b04852 (2018).

    Google Scholar 

  46. Ibrahim, W. W. et al. Neuroprotective potential of Erigeron bonariensis ethanolic extract against ovariectomized/D-galactose-induced memory impairments in female rats in relation to its metabolite fingerprint as revealed using UPLC/MS. Inflammopharmacology 32, 1091–1112. https://doi.org/10.1007/s10787-023-01418-3 (2024).

    Google Scholar 

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Authors and Affiliations

  1. North Henan Medical University, Xinxiang, 453000, China

    Huiqin Qian, Menglin Wang, Haibo Xu, Kun Feng, Yaxuan Li, Yingfan Hu & Yanling Li

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  1. Huiqin Qian
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  2. Menglin Wang
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Contributions

Huiqin Qian : Writing – review & editing, Writing – original draft, Methodology, Formal analysis, Conceptualization. Menglin Wang : Supervision, Resources, Conceptualization. Haibo Xu and Kun Feng : Methodology, Formal analysis. Yaxuan Li and Yingfan Hu : Supervision, Conceptualization. Yanling Li : Writing – review & editing, Methodology.

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Correspondence to Yanling Li.

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Qian, H., Wang, M., Xu, H. et al. Ultrasound-assisted aqueous two-phase extraction of flavonoids from erigeron breviscapus: process optimization, structural characterization, antioxidant study, and DFT calculation. Sci Rep (2026). https://doi.org/10.1038/s41598-026-41556-6

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  • Received: 26 November 2025

  • Accepted: 20 February 2026

  • Published: 03 March 2026

  • DOI: https://doi.org/10.1038/s41598-026-41556-6

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Keywords

  • Erigeron breviscapus
  • Flavonoids
  • Ultrasonic
  • Response surface methodology
  • Density functional theory
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