{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:08:41Z","timestamp":1792789721120,"version":"4.5.0"},"reference-count":60,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T00:00:00Z","timestamp":1789430400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"METROFOOD-CZ Research Infrastructure"},{"name":"Ministry of Education, Youth and Sports of the Czech Republic","award":["LM2023064"],"award-info":[{"award-number":["LM2023064"]}],"id":[{"id":"https:\/\/ror.org\/037n8p820","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The development of sustainable and efficient sample-preparation strategies remains a key challenge in analytical workflows for plant-derived bioactive compounds. In this study, choline chloride (ChCl)-based natural deep eutectic solvents (NADES) were systematically investigated as green extraction media and combined with chemical and enzyme assays to identify the bioactive effect of phenolic compounds from Bellevalia dubia, an understudied Mediterranean plant. A design of experiments approach was applied to evaluate the influence of hydrogen bond donors (lactic, malic, and citric acid) and their molar ratios on extraction performance. Among the tested systems, the ChCl\u2013malic acid (MA) mixture at a molar ratio of 1:2.5 (precisely 1:2.47) was identified as the best overall solution according to the desirability approach, which integrated both phytochemical and enzyme-inhibitory responses. This system also showed strong enzyme-inhibitory effects compared to conventional extractants (water and 80% methanol). To identify the extract composition, a suspect screening workflow was applied based on ultra-high-performance liquid chromatography hybrid quadrupole Orbitrap mass spectrometry (UHPLC\u2013q-Orbitrap-MS). Twenty-one compounds were detected and annotated, mainly belonging to the flavonoid class. Principal component analysis (PCA) showed that ChCl-MA extracts were discriminated, particularly at molar ratio 1:2.5, from the rest, indicating a distinct chemical and bioactivity profile. PCA further suggested a possible association of the ChCl-MA 1:2.5 system with vitexin and AChE inhibition, which requires further investigation. Overall, the proposed NADES-based approach offers a tunable, promising, and potentially more sustainable alternative for the extraction of bioactive compounds from plant-based sources.<\/jats:p>","DOI":"10.3390\/molecules31183267","type":"journal-article","created":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T13:33:42Z","timestamp":1789479222000},"page":"3267","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Experimental Design-Driven Optimization of Choline-Based Natural Deep Eutectic Solvents for the Extraction of Bioactive Compounds from Bellevalia dubia"],"prefix":"10.3390","volume":"31","author":[{"given":"Anastasia","family":"Charalampidopoulou","sequence":"first","affiliation":[{"name":"Laboratory of Food Chemistry and Analysis, Department of Food Science and Human Nutrition, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece"},{"name":"Department of Food Analysis and Nutrition, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Technick\u00e1 5, 166 28 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9920-6110","authenticated-orcid":false,"given":"Marie","family":"Zlechovcov\u00e1","sequence":"additional","affiliation":[{"name":"Department of Food Analysis and Nutrition, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Technick\u00e1 5, 166 28 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3450-5969","authenticated-orcid":false,"given":"Charalampos","family":"Proestos","sequence":"additional","affiliation":[{"name":"Food Chemistry Laboratory, Department of Chemistry, National and Kapodistrian University of Athens, 15772 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1590-688X","authenticated-orcid":false,"given":"Chrysavgi","family":"Gardeli","sequence":"additional","affiliation":[{"name":"Laboratory of Food Chemistry and Analysis, Department of Food Science and Human Nutrition, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4450-2611","authenticated-orcid":false,"given":"Aristeidis S.","family":"Tsagkaris","sequence":"additional","affiliation":[{"name":"Department of Food Analysis and Nutrition, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Technick\u00e1 5, 166 28 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"software","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"169208","DOI":"10.1016\/j.scitotenv.2023.169208","article-title":"Rise, fall and hope for the Sicilian endemic plant Muscari gussonei: A story of survival in the face of narrow germination optimum, climate changes, desertification and habitat fragmentation","volume":"912","author":"Bonanno","year":"2024","journal-title":"Sci. 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