{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T12:01:44Z","timestamp":1790596904934,"version":"4.1.0"},"reference-count":32,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T00:00:00Z","timestamp":1790380800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82204968"],"award-info":[{"award-number":["82204968"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["82204968"],"award-info":[{"award-number":["82204968"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Science and Technology Projects of Guangzhou","award":["2023A04J1153"],"award-info":[{"award-number":["2023A04J1153"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Hovenia dulcis Thunb. is a traditional medicinal and edible plant that is widely recognized for its hepatoprotective and hangover-alleviating properties, which are largely attributed to its rich flavonoid composition. The aim of this study was to develop an environmentally friendly ultrasonic-assisted deep eutectic solvent (UAE-DES) extraction method for the efficient extraction of bioactive flavonoids from Hovenia dulcis Thunb. A Box\u2013Behnken design (BBD) was employed to systematically optimize the extraction parameters. Under optimal conditions (water content of 36%, liquid-to-solid ratio of 42:1 mL\/g, extraction time of 42 min), the extraction yield reached a maximum of 9.09 \u00b1 0.04%. Following purification using AB-8 macroporous resin, the phytochemicals were comprehensively characterized by UPLC-Q-Exactive Orbitrap-HRMS. A total of 12 flavonoids were identified. Bioassays demonstrated that the purified flavonoids exhibited strong, dose-dependent antioxidant activity against DPPH and ABTS radicals in vitro. Furthermore, the extract demonstrated significant anti-inflammatory activity by markedly inhibiting the excessive production of nitric oxide (NO) in RAW 264.7 macrophages induced by lipopolysaccharide (LPS), whilst exhibiting no cytotoxicity within the tested concentration range (0.06\u20130.96 mg\/mL). These findings strongly confirm that the optimized UAE-DES method is a sustainable and efficient strategy for extracting structurally intact and pharmacologically active flavonoids from Hovenia dulcis Thunb, highlighting its immense potential for future applications in the functional food and pharmaceutical industries.<\/jats:p>","DOI":"10.3390\/molecules31193425","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T11:35:59Z","timestamp":1790595359000},"page":"3425","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Ultrasound-Assisted Deep Eutectic Solvent Extraction of Flavonoids from Hovenia dulcis Thunb: Optimization, Phytochemical Profiling, and Antioxidant and Anti-Inflammatory Activity Evaluation"],"prefix":"10.3390","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1332-5619","authenticated-orcid":false,"given":"Lei","family":"Zhou","sequence":"first","affiliation":[{"name":"School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"given":"Haixin","family":"Li","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Xu","family":"Zhou","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"given":"Xueying","family":"Zou","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Guanfeng","family":"He","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6715","DOI":"10.1021\/acs.jafc.0c01117","article-title":"Fruit of Hovenia dulcis Thunb. Induces Nonshivering Thermogenesis through Mitochondrial Biogenesis and Activation by SIRT1 in High-Fat Diet-Fed Obese Mice and Primary Cultured Brown Adipocytes","volume":"68","author":"Song","year":"2020","journal-title":"J. Agric. Food Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.biopha.2017.03.077","article-title":"Anti-steatotic and anti-inflammatory effects of Hovenia dulcis Thunb. extracts in chronic alcohol-fed rats","volume":"90","author":"Choi","year":"2017","journal-title":"Biomed. Pharmacother."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"116042","DOI":"10.1016\/j.jep.2022.116042","article-title":"Protective effect of Hovenia dulcis Thunb. leaf extracts against ethanol-induced DNA damage in SH-SY5Y cells","volume":"304","author":"Garcia","year":"2023","journal-title":"J. Ethnopharmacol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Jegal, K.H., Park, H.R., Choi, B.R., Kim, J.K., and Ku, S.K. (2023). Synergistic Protective Effect of Fermented Schizandrae Fructus Pomace and Hoveniae Semen cum Fructus Extracts Mixture in the Ethanol-Induced Hepatotoxicity. Antioxidants, 12.","DOI":"10.3390\/antiox12081602"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Paik, D.H., Lee, K.W., Shim, Y.Y., Reaney, M.J.T., Park, I., Lee, S.H., Park, J.Y., Park, E., Lee, S.B., and Kim, I.A. (2024). Efficacy of Hovenia dulcis Fruit Extract in Hangover Mitigation: Double-Blind Randomized Clinical Evaluation. Foods, 13.","DOI":"10.3390\/foods13244084"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1711","DOI":"10.1039\/D5FO03575A","article-title":"Dihydromyricetin in the management of diabetes and its complications: A narrative review","volume":"17","author":"Jiang","year":"2026","journal-title":"Food Funct."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1016\/j.ijbiomac.2022.03.211","article-title":"Isolation, purification, structural characterization, and hypoglycemic activity assessment of polysaccharides from Hovenia dulcis (Guai Zao)","volume":"208","author":"Yang","year":"2022","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"147390","DOI":"10.1016\/j.ijbiomac.2025.147390","article-title":"Quercetin-Hovenia dulcis polysaccharide complexes inhibit alpha-glucosidase and mitigate hyperglycemia","volume":"327","author":"Zhu","year":"2025","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e02088","DOI":"10.1002\/cbdv.202502088","article-title":"Total Flavonoids of Hovenia dulcis Seeds Ameliorate Hepatic Fibrosis via Modulating PI3K\/AKT Signaling Pathway In Vitro and In Vivo","volume":"22","author":"Yang","year":"2025","journal-title":"Chem. Biodivers."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"695","DOI":"10.2147\/DDDT.S150284","article-title":"Hot-water extract of the branches of Hovenia dulcis Thunb. (Rhamnaceae) ameliorates low-fiber diet-induced constipation in rats","volume":"12","author":"Choi","year":"2018","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"131097","DOI":"10.1016\/j.ijbiomac.2024.131097","article-title":"A review on extraction, purification, structural characteristics, biological activities, applications of polysaccharides from Hovenia dulcis Thunb. (Guai Zao)","volume":"265","author":"Zhang","year":"2024","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Liga, S., Paul, C., and Peter, F. (2023). Flavonoids: Overview of Biosynthesis, Biological Activity, and Current Extraction Techniques. Plants, 12.","DOI":"10.3390\/plants12142732"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"111268","DOI":"10.1016\/j.foodres.2022.111268","article-title":"A review of ultrasound-assisted extraction for plant bioactive compounds: Phenolics, flavonoids, thymols, saponins and proteins","volume":"157","author":"Yusoff","year":"2022","journal-title":"Food Res. Int."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e70057","DOI":"10.1111\/1541-4337.70057","article-title":"Natural deep eutectic solvents for anthocyanin extraction from agricultural sources: Process parameters, economic and environmental analysis, and industrial challenges","volume":"23","author":"Pires","year":"2024","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Li, D. (2022). Natural deep eutectic solvents in phytonutrient extraction and other applications. Front. Plant Sci., 13.","DOI":"10.3389\/fpls.2022.1004332"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"32982","DOI":"10.1021\/acs.jafc.5c11656","article-title":"Natural Deep Eutectic Solvents (NADES) in Food Systems: Emerging Applications, Extraction Efficiency, Safety Concerns, and Regulatory Challenges","volume":"73","author":"Martins","year":"2025","journal-title":"J. Agric. Food Chem."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"El Maaiden, E., El Kahia, H., Nasser, B., Moustaid, K., Qarah, N., Boukcim, H., Hirich, A., Kouisni, L., and El Kharrassi, Y. (2023). Deep eutectic solvent-ultrasound assisted extraction as a green approach for enhanced extraction of naringenin from Searsia tripartita and retained their bioactivities. Front. Nutr., 10.","DOI":"10.3389\/fnut.2023.1193509"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1111\/1750-3841.16849","article-title":"Ultrasound-assisted deep eutectic solvent extraction of bioactive compounds from persimmon calyx","volume":"89","author":"Kutlu","year":"2024","journal-title":"J. Food Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"127443","DOI":"10.1016\/j.talanta.2024.127443","article-title":"Deep eutectic solvent-based green extraction of Strychnos potatorum seed phenolics: Process optimization via response surface methodology and artificial neural network","volume":"286","author":"Iftikhar","year":"2025","journal-title":"Talanta"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"107434","DOI":"10.1016\/j.ultsonch.2025.107434","article-title":"Optimization of ultrasound-assisted deep eutectic solvent extraction, characterization, and bioactivities of flavonoids from Cercis glabra leaves","volume":"120","author":"Meng","year":"2025","journal-title":"Ultrason. Sonochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"107254","DOI":"10.1016\/j.ultsonch.2025.107254","article-title":"Ultrasound-assisted deep eutectic solvent extraction of flavonol glycosides from Ginkgo biloba: Optimization of efficiency and mechanism","volume":"114","author":"Lai","year":"2025","journal-title":"Ultrason. Sonochem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"143885","DOI":"10.1016\/j.foodchem.2025.143885","article-title":"Enhanced deep eutectic solvent extraction of phenolic compounds from hawthorn seeds with ultrasonic and resin in-situ adsorption assistance: Process optimization, component analysis, and extraction mechanism","volume":"480","author":"Jiao","year":"2025","journal-title":"Food Chem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Bajkacz, S., Adamek, J., and Sobska, A. (2020). Application of Deep Eutectic Solvents and Ionic Liquids in the Extraction of Catechins from Tea. Molecules, 25.","DOI":"10.3390\/molecules25143216"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"19169","DOI":"10.1021\/acssuschemeng.0c08146","article-title":"Insight into the Deep Eutectic Solvent Extraction Mechanism of Flavonoids from Natural Plant","volume":"8","author":"Cao","year":"2020","journal-title":"ACS Sustain. Chem. Eng."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Yu, P., Wang, H., Wang, X., Bai, T., Kang, C., Zhou, J., and Fang, L. (2026). Microwave-assisted natural deep eutectic solvents extraction of saponins from the fibrous roots of American ginseng: Process optimization, antioxidant activity, and metabolomics analysis. npj Sci. Food, Online ahead of print.","DOI":"10.1038\/s41538-026-00983-5"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Sferrazza, G., Brusotti, G., Zonfrillo, M., Temporini, C., Tengattini, S., Bononi, M., Tateo, F., Calleri, E., and Pierimarchi, P. (2021). Hovenia dulcis Thumberg: Phytochemistry, Pharmacology, Toxicology and Regulatory Framework for Its Use in the European Union. Molecules, 26.","DOI":"10.3390\/molecules26040903"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e10097","DOI":"10.1002\/rcm.10097","article-title":"Phytochemical Analysis of Semen Hoveniae and Its Potential Mechanism Against Alcoholic Liver Disease via an Integrated Approach Combining UHPLC-Q-TOF MS, Network Pharmacology, and Molecular Docking","volume":"39","author":"Li","year":"2025","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_28","first-page":"130","article-title":"HPLC Determination of Bioactive Flavonoids in Hovenia dulcis Fruit Extracts","volume":"54","author":"Park","year":"2016","journal-title":"J. Chromatogr. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e24435","DOI":"10.1016\/j.heliyon.2024.e24435","article-title":"Phytochemical screening, antioxidant activity of selected methanolic plant extracts and their detoxification capabilities against AFB(1) toxicity","volume":"10","author":"Makhuvele","year":"2024","journal-title":"Heliyon"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Martinez, S., Fuentes, C., and Carballo, J. (2022). Antioxidant Activity, Total Phenolic Content and Total Flavonoid Content in Sweet Chestnut (Castanea sativa Mill.) Cultivars Grown in Northwest Spain under Different Environmental Conditions. Foods, 11.","DOI":"10.3390\/foods11213519"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"9184769","DOI":"10.1155\/2019\/9184769","article-title":"Hoveniae Semen Seu Fructus Ethanol Extract Exhibits Anti-Inflammatory Activity via MAPK, AP-1, and STAT Signaling Pathways in LPS-Stimulated RAW 264.7 and Mouse Peritoneal Macrophages","volume":"2019","author":"Jeong","year":"2019","journal-title":"Mediat. Inflamm."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"113888","DOI":"10.1016\/j.bcp.2020.113888","article-title":"Dihydromyricetin alleviates doxorubicin-induced cardiotoxicity by inhibiting NLRP3 inflammasome through activation of SIRT1","volume":"175","author":"Sun","year":"2020","journal-title":"Biochem. Pharmacol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/19\/3425\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T11:37:28Z","timestamp":1790595448000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/19\/3425"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,26]]},"references-count":32,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2026,10]]}},"alternative-id":["molecules31193425"],"URL":"https:\/\/doi.org\/10.3390\/molecules31193425","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,26]]}}}