{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T14:06:16Z","timestamp":1787753176309,"version":"build-2784847793"},"reference-count":83,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T00:00:00Z","timestamp":1783036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000780","name":"European Union","doi-asserted-by":"crossref","award":["BG-RRP-2.017-0043\/16.12.2024"],"award-info":[{"award-number":["BG-RRP-2.017-0043\/16.12.2024"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"crossref"}]},{"award":["BG-RRP-2.017-0043\/16.12.2024"],"award-info":[{"award-number":["BG-RRP-2.017-0043\/16.12.2024"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>This study investigates subcritical water extraction (SWE) as an alternative to hydroalcoholic extraction for the production of Echinacea purpurea root extracts standardized to hydroxycinnamic acids (cichoric and caftaric acids). Extractions were performed at 100 \u00b0C, 125 \u00b0C, 150 \u00b0C, and 170 \u00b0C for 10\u201330 min. The recovery of cichoric and caftaric acids was significantly (p &lt; 0.05) influenced by extraction temperature, with the highest values obtained within the range of 100\u2013125 \u00b0C. Further experiments identified 110 \u00b0C for 10 min as the optimal condition, yielding the highest cumulative recovery of cichoric and caftaric acids (1.87 \u00b1 0.10% of dry material). In the resulting dry extracts, SWE at 100\u2013125 \u00b0C produced hydroxycinnamic acid contents of 5.5\u20137.1%, whereas the total dry extract yield in-creased from 24\u201328% at 100 \u00b0C to 40\u201341% at 150\u2013170 \u00b0C (p &lt; 0.05). Higher temperatures, however, reduced cichoric and caftaric acid cumulative content to 0.6\u20131.7% (p &lt; 0.05), indicating a degradation of the target compounds. In contrast, total polyphenol recovery in-creased continuously with temperature, reaching 4.86% at 170 \u00b0C for 30 min. This was accompanied by marked increases in rutin, gallic and caffeic acid, reaching 458.5 mg\/100 g dry weight (DW), 175.5 mg\/100 g DW and 945.7 mg\/100 g DW (p &lt; 0.05), respectively, suggesting the release of bound phenolics following partial disruption of plant cell wall structures. SWE also enhanced the extraction of carbohydrates, uronic acids, fructans, proteins and organic acids, demonstrating an extensive temperature-dependent modification of the root matrix. 5-HMF was not detected in extracts obtained below 125 \u00b0C, but increased progressively at higher temperatures, reaching 200 mg\/100 g (p &lt; 0.05) at 170 \u00b0C. Biological evaluation in the human colorectal adenocarcinoma cell line (HT29) showed favorable cytocompatibility of SWE extracts, confirmed by cell viability, morphological assessment and low DNA damage in the Comet Assay. Overall, SWE enables the production of cichoric and caftaric acid-standardized E. purpurea extracts without organic solvents, supporting its application in pharmaceutical, nutraceutical, food and cosmeceutical products.<\/jats:p>","DOI":"10.3390\/molecules31132351","type":"journal-article","created":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T11:39:11Z","timestamp":1783078751000},"page":"2351","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Subcritical Water Extraction Enables the Production of Cichoric and Caftaric Acid-Standardized Echinacea purpurea Root Extracts with Defined Composition and Favorable Biological Properties"],"prefix":"10.3390","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2290-791X","authenticated-orcid":false,"given":"Petko","family":"Denev","sequence":"first","affiliation":[{"name":"Laboratory of Biologically Active Substances, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 4000 Plovdiv, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3109-2238","authenticated-orcid":false,"given":"Desislava","family":"Teneva","sequence":"additional","affiliation":[{"name":"Laboratory of Biologically Active Substances, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 4000 Plovdiv, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5568-0854","authenticated-orcid":false,"given":"Manol","family":"Ognyanov","sequence":"additional","affiliation":[{"name":"Laboratory of Biologically Active Substances, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 4000 Plovdiv, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6105-2715","authenticated-orcid":false,"given":"Mariya","family":"Pimpilova","sequence":"additional","affiliation":[{"name":"Laboratory of Biologically Active Substances, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 4000 Plovdiv, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ani","family":"Petrova","sequence":"additional","affiliation":[{"name":"Laboratory of Biologically Active Substances, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 4000 Plovdiv, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georgi","family":"Dimitrov","sequence":"additional","affiliation":[{"name":"Laboratory of Biologically Active Substances, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 4000 Plovdiv, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8718-6412","authenticated-orcid":false,"given":"Bela","family":"Vasileva","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology \u201cAcad. R. Tsanev\u201d, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8348-8336","authenticated-orcid":false,"given":"Kamelia","family":"Hristova-Panusheva","sequence":"additional","affiliation":[{"name":"Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7423-7412","authenticated-orcid":false,"given":"Natalia","family":"Krasteva","sequence":"additional","affiliation":[{"name":"Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2979-8899","authenticated-orcid":false,"given":"George","family":"Miloshev","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology \u201cAcad. R. Tsanev\u201d, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2371-7544","authenticated-orcid":false,"given":"Milena","family":"Georgieva","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology \u201cAcad. R. Tsanev\u201d, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Puchalski, K., Gerstel, J.A., Jimoh, A., Shokoohinia, Y., and Langland, J. (2025). Effects of Echinacea purpurea and Alkylamides on Respiratory Virus Replication and IL-8 Expression In Vitro. Molecules, 30.","DOI":"10.3390\/molecules30020386"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.ctim.2019.03.011","article-title":"Echinacea for the Prevention and Treatment of Upper Respiratory Tract Infections: A Systematic Review and Meta-Analysis","volume":"44","author":"David","year":"2019","journal-title":"Complement. Ther. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"100397","DOI":"10.1016\/j.prmcm.2024.100397","article-title":"Echinacea spp.: The cold-fighter herbal remedy?","volume":"10","author":"Kakouri","year":"2024","journal-title":"Pharmacol. Res. Mod. Chin. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"769896","DOI":"10.1155\/2012\/769896","article-title":"Applications of the Phytomedicine Echinacea purpurea (Purple Coneflower) in Infectious Diseases","volume":"2012","author":"Hudson","year":"2012","journal-title":"J. Biomed. Biotechnol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/S0006-2952(99)00413-X","article-title":"Use of echinacea in medicine","volume":"60","author":"Percival","year":"2000","journal-title":"Biochem. Pharmacol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ahmadi, F. (2024). Phytochemistry, Mechanisms, and Preclinical Studies of Echinacea Extracts in Modulating Immune Responses to Bacterial and Viral Infections: A Comprehensive Review. Antibiotics, 13.","DOI":"10.3390\/antibiotics13100947"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Espinoza, J.P., Guajardo, V., Rodr\u00edguez-D\u00edaz, M., Moreno, M., Klagges, C., Castillo-Ruiz, M., and Otero, M.C. (2026). Pharmacological Insights and Technological Innovations in Curcuma longa L. and Echinacea purpurea (L.) Moench as Plant-Derived Immunomodulators. Pharmaceuticals, 19.","DOI":"10.3390\/ph19010093"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Burlou-Nagy, C., B\u0103nic\u0103, F., Negrean, R.A., Jurca, T., Vica\u0219, L.G., Marian, E., B\u00e1cskay, I., Kiss, R., Feh\u00e9r, P., and Vica\u0219, S.I. (2023). Determination of the Bioactive Compounds from Echinacea purpurea (L.) Moench Leaves Extracts in Correlation with the Antimicrobial Activity and the In Vitro Wound Healing Potential. Molecules, 28.","DOI":"10.3390\/molecules28155711"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Burlou-Nagy, C., B\u0103nic\u0103, F., Jurca, T., Vica\u0219, L.G., Marian, E., Muresan, M.E., B\u00e1cskay, I., Kiss, R., Feh\u00e9r, P., and Pallag, A. (2022). Echinacea purpurea (L.) Moench: Biological and Pharmacological Properties. A Review. Plants, 11.","DOI":"10.3390\/plants11091244"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1653","DOI":"10.1002\/ptr.6101","article-title":"Echinacea Plants as Antioxidant and Antibacterial Agents: From Traditional Medicine to Biotechnological Applications","volume":"32","author":"Mnayer","year":"2018","journal-title":"Phytother. Res."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Petrova, A., Ognyanov, M., Petkova, N., and Denev, P. (2023). Phytochemical Characterization of Purple Coneflower Roots (Echinacea purpurea (L.) Moench.) and Their Extracts. Molecules, 28.","DOI":"10.3390\/molecules28093956"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.jep.2015.09.007","article-title":"Echinacea Complex\u2013Chemical View and Anti-Asthmatic Profile","volume":"175","author":"Capek","year":"2015","journal-title":"J. Ethnopharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"109","DOI":"10.2478\/jvetres-2021-0016","article-title":"Echinacea Purpurea Extract (Cichoric Acid) Exerts an Anti-Inflammatory Effect on Yak PBMCs and Regulates the TLR4 Signalling Pathway","volume":"65","author":"Xue","year":"2021","journal-title":"J. Vet. Res."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Vieira, S.F., Gon\u00e7alves, S.M., Gon\u00e7alves, V.M.F., Llaguno, C.P., Mac\u00edas, F., Tiritan, M.E., Cunha, C., Carvalho, A., Reis, R.L., and Ferreira, H. (2023). Echinacea purpurea Fractions Represent Promising Plant-Based Anti-Inflammatory Formulations. Antioxidants, 12.","DOI":"10.3390\/antiox12020425"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Ciganovi\u0107, P., Jakupovi\u0107, L., Momchev, P., Ni\u017ei\u0107 Nodilo, L., Hafner, A., and Zovko Kon\u010di\u0107, M. (2023). Extraction Optimization, Antioxidant, Cosmeceutical and Wound Healing Potential of Echinacea purpurea Glycerolic Extracts. Molecules, 28.","DOI":"10.3390\/molecules28031177"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.foodchem.2005.11.048","article-title":"Antioxidant activity of cichoric acid and alkamides from Echinacea purpurea, alone and in combination","volume":"101","author":"Thygesen","year":"2007","journal-title":"Food Chem."},{"key":"ref_17","unstructured":"European Pharmacopoeia Commission (2011). 01\/2008:1824. Purple coneflower root. European Pharmacopoeia, Council of Europe. [7th ed.]."},{"key":"ref_18","unstructured":"United States Pharmacopeial Convention (2020). United States Pharmacopeia and National Formulary (USP 43\u2013NF 38), USP Convention."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.cofs.2021.02.009","article-title":"Conventional versus green extraction techniques\u2014A comparative perspective","volume":"40","author":"Mahomoodally","year":"2021","journal-title":"Curr. Opin. Food Sci."},{"key":"ref_20","first-page":"100184","article-title":"Sustainable Green Extraction Approaches for Herbal Phytochemicals Supporting Environmentally Friendly and Climate Responsive Product Development","volume":"8","author":"Plyduang","year":"2026","journal-title":"Sustain. Chem. Clim. Action"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Sun, S., Yu, Y., Jo, Y., Han, J.H., Xue, Y., Cho, M., Bae, S.J., Ryu, D., Park, W., and Ha, K.T. (2025). Impact of Extraction Techniques on Phytochemical Composition and Bioactivity of Natural Product Mixtures. Front. Pharmacol., 16.","DOI":"10.3389\/fphar.2025.1615338"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1080\/87559129.2020.1772283","article-title":"An Inclusive Overview of Advanced Thermal and Nonthermal Extraction Techniques for Bioactive Compounds in Food and Food-Related Matrices","volume":"38","author":"Zia","year":"2022","journal-title":"Food Rev. Int."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1080\/10408398.2021.1952923","article-title":"Recent Advances in Extraction Technologies for Recovery of Bioactive Compounds Derived from Fruit and Vegetable Waste Peels: A Review","volume":"63","author":"Rifna","year":"2023","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_24","first-page":"100399","article-title":"Optimization of heat- and ultrasound-assisted extraction of bioactive compounds from Echinacea purpurea using response surface methodology","volume":"31","author":"Sobhani","year":"2022","journal-title":"J. Appl. Res. Med. Aromat. Plants"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"117201","DOI":"10.1016\/j.trac.2023.117201","article-title":"Pressurized hot water extraction of bioactives","volume":"166","author":"Plaza","year":"2023","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.aca.2011.07.018","article-title":"Pressurized liquid extraction as a green approach in food and herbal plants","volume":"703","author":"Mustafa","year":"2011","journal-title":"Anal. Chim. Acta"},{"key":"ref_27","unstructured":"\u017dagar, T., Hrn\u010di\u010d, M.K., \u0160kerget, M., Knez, \u017d., and Bren, U. (2024). Polyphenol-rich extracts obtained by subcritical water extraction: Composition and antioxidant activity. Foods, 13."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/S0021-9673(02)01225-6","article-title":"Extractions with superheated water","volume":"975","author":"Smith","year":"2002","journal-title":"J. Chromatogr. A"},{"key":"ref_29","unstructured":"Monagas, M., Urpi-Sarda, M., S\u00e1nchez-Pat\u00e1n, F., Llorach, R., Garrido, I., G\u00f3mez-Cordov\u00e9s, C., Andres-Lacueva, C., and Bartolom\u00e9, B. (2022). Insights into plant extract standardization and bioactive compounds. Nutrients, 14."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"213","DOI":"10.47583\/ijpsrr.2021.v68i01.033","article-title":"Standardization of herbal drugs\u2014An overview","volume":"68","author":"Umamaheswari","year":"2021","journal-title":"Int. J. Pharm. Sci. Rev. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"100211","DOI":"10.1016\/j.medidd.2025.100211","article-title":"Current demands for standardization of Indian medicinal plants: A critical review","volume":"27","author":"Kaundal","year":"2025","journal-title":"Med. Drug Discov."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"509S","DOI":"10.1093\/ajcn\/87.2.509S","article-title":"Ensuring the safety and quality of botanical dietary supplements","volume":"87","author":"Fong","year":"2008","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_33","first-page":"1017","article-title":"Challenges in standardization of herbal medicines","volume":"58","author":"Srinivasan","year":"2006","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Senica, M., Mlin\u0161ek, G., Rusjan, D., and Mikulic-Petkovsek, M. (2025). Innovative Approaches to Extracting Phenolics from Echinacea purpurea: Maximizing Yield and Efficacy. Foods, 14.","DOI":"10.3390\/foods14132325"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.supflu.2016.09.002","article-title":"Solid\u2013liquid and high-pressure (liquid and supercritical carbon dioxide) extraction of Echinacea purpurea L.","volume":"119","year":"2017","journal-title":"J. Supercrit. Fluids"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"107507","DOI":"10.1016\/j.microc.2022.107507","article-title":"Chromatographic analysis of water and water\u2013alcohol extracts of Echinacea purpurea L. obtained by various methods","volume":"179","author":"Temerdashev","year":"2022","journal-title":"Microchem. J."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Vlasheva, M., Katsarova, M., Dobreva, A., Dzhurmanski, A., Denev, P., and Dimitrova, S. (2024). Echinacea Species Cultivated in Bulgaria as a Source of Chicoric and Caftaric Acids. Agronomy, 14.","DOI":"10.3390\/agronomy14092081"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Petkova, N., Petrova, A., Ivanov, I., Hambarlyiska, I., Tumbarski, Y., Dincheva, I., Ognyanov, M., and Denev, P. (2023). Chemical Composition of Different Extracts from Echinacea purpurea (L.) Moench Roots and Evaluation of Their Antimicrobial Activity. ChemEngineering, 7.","DOI":"10.3390\/chemengineering7050094"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1134\/S199079311308006X","article-title":"Extraction of caftaric and cichoric acids from Echinacea purpurea L. in subcritical water","volume":"7","author":"Lekar","year":"2013","journal-title":"Russ. J. Phys. Chem. B"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Abeynayake, S.W., Etzerodt, T.P., Jonavi\u010dien\u0117, K., Byrne, S., Asp, T., and Boelt, B. (2015). Fructan metabolism and changes in fructan composition during cold acclimation in perennial ryegrass. Front. Plant Sci., 6.","DOI":"10.3389\/fpls.2015.00329"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Yoshida, M. (2021). Fructan Structure and Metabolism in Overwintering Plants. Plants, 10.","DOI":"10.3390\/plants10050933"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Denev, P., Ognyanov, M., Pimpilova, M., and Teneva, D. (2026). Subcritical Water Extraction as a Green Technology for the Development of Standardized Plant Extracts for Food and Pharmaceutical Uses. Processes, 14.","DOI":"10.3390\/pr14101564"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.jfoodeng.2010.09.008","article-title":"Subcritical water extraction of flavonol quercetin from onion skin","volume":"102","author":"Ko","year":"2011","journal-title":"J. Food Eng."},{"key":"ref_44","first-page":"37","article-title":"Extraction and characterization of bioactive compounds with subcritical water","volume":"1250","author":"Herrero","year":"2012","journal-title":"J. Chromatogr. A"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1016\/j.jpba.2018.11.027","article-title":"Design of experiments in metabolomics-related studies: An overview","volume":"164","author":"Jacyna","year":"2019","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1945","DOI":"10.1021\/jf404850a","article-title":"Degradation of caffeic acid in subcritical water and online HPLC\u2013DPPH assay of degradation products","volume":"62","author":"Khuwijitjaru","year":"2014","journal-title":"J. Agric. Food Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.supflu.2014.02.005","article-title":"A bioactivity-based comparison of Echinacea purpurea extracts obtained by various processes","volume":"89","author":"Yildiz","year":"2014","journal-title":"J. Supercrit. Fluids"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.trac.2015.02.022","article-title":"Pressurized hot water extraction of bioactives","volume":"71","author":"Plaza","year":"2015","journal-title":"Trends Anal. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"104561","DOI":"10.1016\/j.supflu.2019.104561","article-title":"Subcritical water extraction of rutin from the aerial parts of common buckwheat","volume":"152","author":"Kim","year":"2019","journal-title":"J. Supercrit. Fluids"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"10920","DOI":"10.1021\/jf3027759","article-title":"Effects of temperature and time on polyphenolic content and antioxidant activity in the pressurized hot water extraction of deodorized thyme (Thymus vulgaris)","volume":"60","author":"Torres","year":"2012","journal-title":"J. Agric. Food Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1211","DOI":"10.1080\/13880209.2023.2244000","article-title":"A standardized extract of Echinacea purpurea containing higher cichoric acid content enhances immune function in murine macrophages and cyclophosphamide-induced immunosuppression mice","volume":"61","author":"Sudeep","year":"2023","journal-title":"Pharm. Biol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"351","DOI":"10.15255\/CABEQ.2015.2193","article-title":"Hydrothermal degradation of fats, carbohydrates and proteins in sunflower seeds after treatment with subcritical water","volume":"29","author":"Ravber","year":"2015","journal-title":"Chem. Biochem. Eng. Q."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1637","DOI":"10.1007\/s13399-022-02380-w","article-title":"Optimization of the subcritical water treatment from sunflower by-product for producing protein and sugar extracts","volume":"14","author":"Alonso","year":"2024","journal-title":"Biomass Convers. Biorefinery"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Zong, X., Lei, N., Yin, J., He, W., Nie, S., and Xie, M. (2024). Exploration and improvement of acid hydrolysis conditions for inulin-type fructans monosaccharide composition analysis: Monosaccharide recovery and by-product identification. Foods, 13.","DOI":"10.3390\/foods13081241"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2883","DOI":"10.1021\/ie990690j","article-title":"Dissolution and hydrolysis of cellulose in subcritical and supercritical water","volume":"39","author":"Sasaki","year":"2000","journal-title":"Ind. Eng. Chem. Res."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"130911","DOI":"10.1016\/j.ijbiomac.2024.130911","article-title":"Solubilization and structural changes of lignin in naked oat stems during subcritical water autohydrolysis","volume":"265","author":"Jiang","year":"2024","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.bej.2003.08.002","article-title":"Hydrolysis of disaccharides containing glucose residue in subcritical water","volume":"18","author":"Oomori","year":"2004","journal-title":"Biochem. Eng. J."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1016\/j.lwt.2004.06.005","article-title":"Kinetics on sucrose decomposition in subcritical water","volume":"38","author":"Khajavi","year":"2005","journal-title":"LWT Food Sci. Technol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/S0008-6215(00)80500-5","article-title":"Acid-catalyzed degradation of d-fructose","volume":"5","author":"Shaw","year":"1967","journal-title":"Carbohydr. Res."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"4947","DOI":"10.1002\/cssc.202101487","article-title":"Hydrothermal degradation of amino acids","volume":"14","year":"2021","journal-title":"ChemSusChem"},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Fan, R., and Gao, Y. (2022). Maillard and Hydrolytic Reactions in Subcritical Water Extraction of Bioactive Compounds from Licorice. Molecules, 27.","DOI":"10.3390\/molecules27206851"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.foodres.2012.01.008","article-title":"Formation and relevance of 5-hydroxymethylfurfural in bioactive subcritical water extracts from olive leaves","volume":"47","author":"Herrero","year":"2012","journal-title":"Food Res. Int."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1007\/s10068-018-0328-y","article-title":"5-Hydroxymethylfurfural (HMF) formation during subcritical water extraction","volume":"27","year":"2018","journal-title":"Food Sci. Biotechnol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"118573","DOI":"10.1016\/j.lwt.2025.118573","article-title":"Polysaccharides recovery by subcritical water as an additional step for a biorefinery valorisation of grape marc with pressurized fluids","volume":"235","author":"Natolino","year":"2025","journal-title":"LWT Food Sci. Technol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"2075614","DOI":"10.1155\/2019\/2075614","article-title":"Medicinal Plants in the Prevention and Treatment of Colon Cancer","volume":"2019","author":"Aiello","year":"2019","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Gerasimova, T., Gateva, S., Jovtchev, G., Dobreva, A., Mileva, M., Kokanova-Nedialkova, Z., Gospodinova, M., Angelova, T., and Nedialkov, P. (2025). Screening of Cytotoxic and Genotoxic Activities of Subcritical Water Extracts from R. amascena and R. alba Flowers. Molecules, 30.","DOI":"10.3390\/molecules30214294"},{"key":"ref_67","unstructured":"(2016). Determination of Protein in Foods. National Food Safety Standard (NFSS) of the People\u2019s Republic of China (Standard No. GB 5009.5-2016)."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1021\/ac60111a017","article-title":"Colorimetric method for determination of sugars and related substances","volume":"28","author":"DuBois","year":"1956","journal-title":"Anal. Chem."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1111\/j.1745-4514.1978.tb00193.x","article-title":"A simplified method for accurate determination of cell wall uronide content","volume":"1","author":"Ahmed","year":"1978","journal-title":"J. Food Biochem."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1016\/0003-2697(73)90377-1","article-title":"New method for quantitative determination of uronic acids","volume":"54","author":"Blumenkrantz","year":"1973","journal-title":"Anal. Biochem."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1016\/S0003-2697(69)80009-6","article-title":"Semimicro determination of cellulose in biological materials","volume":"32","author":"Updegraff","year":"1969","journal-title":"Anal. Biochem."},{"key":"ref_72","first-page":"488","article-title":"Color reactions of hexoses","volume":"Volume 1","author":"Whistler","year":"1962","journal-title":"Methods in Carbohydrate Chemistry"},{"key":"ref_73","first-page":"1876","article-title":"Characterization of inulin from dahlia tubers isolated by microwave and ultrasound-assisted extractions","volume":"25","author":"Petkova","year":"2018","journal-title":"Int. Food Res. J."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1093\/jaoac\/92.1.42","article-title":"Determination of starch, including maltooligosaccharides, in animal feeds: Comparison of methods and a method recommended for AOAC collaborative study","volume":"92","author":"Hall","year":"2009","journal-title":"J. AOAC Int."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.anifeedsci.2014.12.007","article-title":"Comparison of acetyl bromide lignin with acid detergent lignin and Klason lignin and correlation with in vitro forage degradability","volume":"201","author":"Fukushima","year":"2015","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Lin, S.Y., and Dence, C.W. (1992). The determination of lignin. Methods in Lignin Chemistry, Springer.","DOI":"10.1007\/978-3-642-74065-7"},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Atanasova, A., Petrova, A., Teneva, D., Ognyanov, M., Georgiev, Y., Nenov, N., and Denev, P. (2023). Subcritical Water Extraction of Rosmarinic Acid from Lemon Balm (Melissa officinalis L.) and Its Effect on Plant Cell Wall Constituents. Antioxidants, 12.","DOI":"10.3390\/antiox12040888"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"144","DOI":"10.5344\/ajev.1965.16.3.144","article-title":"Colorimetry of total phenolic with phosphomolibdiphosphotungstic acid reagents","volume":"16","author":"Singleton","year":"1965","journal-title":"Am. J. Enol. Vitic."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"4619","DOI":"10.1021\/jf010586o","article-title":"Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescence probe","volume":"49","author":"Ou","year":"2001","journal-title":"J. Agric. Food Chem."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"1055","DOI":"10.1016\/j.foodchem.2010.05.061","article-title":"Solid-phase extraction of berries\u2019 anthocyanins and evaluation of their antioxidative properties","volume":"123","author":"Denev","year":"2010","journal-title":"Food Chem."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"2772","DOI":"10.1021\/jf011480w","article-title":"Novel fluorometric assay for hydroxyl radical prevention capacity using fluorescein as the probe","volume":"50","author":"Ou","year":"2002","journal-title":"J. Agric. Food Chem."},{"key":"ref_82","doi-asserted-by":"crossref","unstructured":"Zembrzuska, J., Pakulski, \u0141., Karbowska, B., Bartoszewicz, J., Janeba-Bartoszewicz, E., Selech, J., and Kurczewski, P. (2024). Comparison of Different Methods for Determining 5-hydroxymethylfurfural in Assessing the Quality of Honey from Greater Poland. Sustainability, 16.","DOI":"10.3390\/su16114568"},{"key":"ref_83","doi-asserted-by":"crossref","unstructured":"Georgieva, M., Gospodinova, Z., Keremidarska-Markova, M., Kamenska, T., Gencheva, G., and Krasteva, N. (2021). PEGylated nanographene oxide in combination with near-infrared laser irradiation as a smart nanocarrier in colon cancer targeted therapy. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13030424"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/13\/2351\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,5]],"date-time":"2026-07-05T04:17:35Z","timestamp":1783225055000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/13\/2351"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,3]]},"references-count":83,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["molecules31132351"],"URL":"https:\/\/doi.org\/10.3390\/molecules31132351","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,3]]}}}