{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T13:29:45Z","timestamp":1777469385682,"version":"3.51.4"},"reference-count":61,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2024,9,2]],"date-time":"2024-09-02T00:00:00Z","timestamp":1725235200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union\u2019s Horizon 2020 research and innovation programme","award":["778168"],"award-info":[{"award-number":["778168"]}]},{"name":"European Union\u2019s Horizon 2020 research and innovation programme","award":["K\u041f-06-H61\/1 13.12.22"],"award-info":[{"award-number":["K\u041f-06-H61\/1 13.12.22"]}]},{"name":"Bulgarian National Science Fund","award":["778168"],"award-info":[{"award-number":["778168"]}]},{"name":"Bulgarian National Science Fund","award":["K\u041f-06-H61\/1 13.12.22"],"award-info":[{"award-number":["K\u041f-06-H61\/1 13.12.22"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Sangre de drago, the sap of Croton lechleri M\u00fcll. Arg. tree, has been used for centuries in traditional medicine owing to its diverse biological activities. Extracts derived from different parts of the species contain a multitude of phytochemicals with varied applications. Twigs, however, are among the least studied parts of the plant. This study unveils new biological activities of Croton lechleri twig extracts recovered by applying Soxhlet and advanced green techniques. For all extracts, total phenolic content and antioxidant activity were determined. Subsequently, four were selected, and their cytotoxic effects were assessed on both normal (HaCat) and malignant melanoma (A375) skin cell lines using the MTT assay and trypan blue exclusion assay. All showed dose-dependent cytotoxicity, with the Soxhlet ethanol extract demonstrating the highest selectivity towards A375 cells over HaCat cells. The extracts induced apoptosis and necrosis, as confirmed by Annexin V\/PI dual-labeling and flow cytometry, highlighting their ability to trigger programmed cell death in cancer cells. The selective inhibition of cell cycle progression in A375 compared to HaCat observed both for Soxhlet ethanol and pressurized ethanol extracts induces cell cycle arrest at multiple points, primarily in the G1 and G2\/M phases, and significantly reduces DNA synthesis as evidenced by the decrease in the S-phase population, confirmed by the EdU assay. Consequently, the Soxhlet extract composition was analyzed using LC-MS, which revealed their richness in polyphenolic compounds, particularly flavonoids from the flavonol subclass.<\/jats:p>","DOI":"10.3390\/molecules29174161","type":"journal-article","created":{"date-parts":[[2024,9,2]],"date-time":"2024-09-02T10:07:35Z","timestamp":1725271655000},"page":"4161","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Novel Insights into the Biological Activity of Croton lechleri Twigs Extracts and Advancements in Their Sustainable Recovery"],"prefix":"10.3390","volume":"29","author":[{"given":"Alexander","family":"Tzintzarov","sequence":"first","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]},{"given":"Stanislava S.","family":"Boyadzhieva","sequence":"additional","affiliation":[{"name":"Institute of Chemical Engineering, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8118-0864","authenticated-orcid":false,"given":"Jose A. P.","family":"Coelho","sequence":"additional","affiliation":[{"name":"Instituto Superior de Engenharia de Lisboa, Instituto Polit\u00e9cnico de Lisboa, Rua Conselheiro Em\u00eddio Navarro 1, 1959-007 Lisboa, Portugal"},{"name":"Centro de Qu\u00edmica Estrutural, Institute of Molecular Sciences, Instituto Superior T\u00e9cnico, Universidade de Lisboa, Av. Rovisco Pais, 1049 001 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3101-8388","authenticated-orcid":false,"given":"Flora","family":"Tsvetanova","sequence":"additional","affiliation":[{"name":"Institute of Chemical Engineering, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0766-3934","authenticated-orcid":false,"given":"Maria","family":"Petrova","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]},{"given":"Georgi","family":"Stoev","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0433-7930","authenticated-orcid":false,"given":"Dragomir S.","family":"Yankov","sequence":"additional","affiliation":[{"name":"Institute of Chemical Engineering, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0116-0079","authenticated-orcid":false,"given":"Iva","family":"Ugrinova","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4834-8183","authenticated-orcid":false,"given":"Roumiana P.","family":"Stateva","sequence":"additional","affiliation":[{"name":"Institute of Chemical Engineering, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}]}],"member":"1968","published-online":{"date-parts":[[2024,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Salazar-G\u00f3mez, A., and Alonso-Castro, A.J. (2022). Medicinal Plants from Latin America with Wound Healing Activity: Ethnomedicine, Phytochemistry, Preclinical and Clinical Studies\u2014A Review. Pharmaceuticals, 15.","DOI":"10.3390\/ph15091095"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/j.jep.2007.10.018","article-title":"Dragon\u2019s blood: Botany, chemistry and therapeutic uses","volume":"115","author":"Gupta","year":"2008","journal-title":"J. Ethnopharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Xu, W.-H., Liu, W.-Y., and Liang, Q. (2018). Chemical Constituents from Croton Species and Their Biological Activities. Molecules, 23.","DOI":"10.3390\/molecules23092333"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1016\/j.jep.2004.08.025","article-title":"Mutagenic and antioxidant activities of Croton lechleri sap in biological systems","volume":"95","author":"Lopez","year":"2004","journal-title":"J. Ethnopharmacol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1016\/j.jep.2012.10.032","article-title":"Croton lechleri sap and isolated alkaloid taspine exhibit inhibition against human melanoma SK23 and colon cancer HT29 cell lines","volume":"144","author":"Montopoli","year":"2012","journal-title":"J. Ethnopharmacol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e57311225306","DOI":"10.33448\/rsd-v11i2.25306","article-title":"Croton sp.: A review about Popular Uses, Biological Activities and Chemical Composition","volume":"11","author":"Ferreira","year":"2022","journal-title":"Res. Soc. Dev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"383","DOI":"10.2174\/1573407215666181122103511","article-title":"Bioactive Compounds and Biological Activity of Croton Species (Euphorbiaceae): An Overview","volume":"16","author":"Bezerra","year":"2020","journal-title":"Curr. Bioact. Compd."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1007\/s12210-022-01122-4","article-title":"Dragon\u2019s Blood: Antioxidant properties for nutraceuticals and pharmaceuticals","volume":"34","author":"Peres","year":"2023","journal-title":"Rend. Lincei-Sci. Fis. Nat."},{"key":"ref_9","first-page":"167","article-title":"Dragon\u2019s blood (Croton lechleri Mull., Arg.): An update on the chemical composition and medical applications of this natural plant extract. A review","volume":"7","author":"Lopez","year":"2013","journal-title":"Rev. Bras. Hig. Sanidade Anim."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4165","DOI":"10.1080\/14786419.2021.1960520","article-title":"Effects of Croton lechleri sap (Sangre de Drago) on AGEs formation, LDL oxidation and oxidative stress related to vascular diseases","volume":"36","author":"Chen","year":"2022","journal-title":"Nat. Prod. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"104992","DOI":"10.1016\/j.supflu.2020.104992","article-title":"Chemical composition, antioxidant activity, anti-inflammatory and neuroprotective effect of Croton matourensis Aubl. Leaves extracts obtained by supercritical CO2","volume":"165","author":"Bezerra","year":"2020","journal-title":"J. Supercrit. Fluids"},{"key":"ref_12","first-page":"471","article-title":"Flavonols and sesquiterpenoids from outer bark and leaves of Croton polycarpus Benth. (Euphorbiaceae)","volume":"16","year":"2017","journal-title":"Bol. Latinoam. Caribe Plantas Med. Aromat."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1016\/j.foodchem.2010.11.042","article-title":"Chemical fingerprinting and bioactivity of Amazonian Ecuador Croton lechleri M\u00fcll. Arg. (Euphorbiaceae) stem bark essential oil: A new functional food ingredient?","volume":"126","author":"Rossi","year":"2011","journal-title":"Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"105525","DOI":"10.1016\/j.microc.2020.105525","article-title":"Greener ultrasound-assisted extraction of bioactive phenolic compounds in Croton heliotropiifolius Kunth leaves","volume":"159","author":"Anjos","year":"2020","journal-title":"Microchem. J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.jfoodeng.2004.08.023","article-title":"Extraction of volatile oil from Croton zehntneri Pax et Hoff with pressurized CO2: Solubility, composition and kinetics","volume":"69","author":"Sousa","year":"2005","journal-title":"J. Food Eng."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"152846","DOI":"10.1016\/j.phymed.2019.152846","article-title":"Supercritical CO2 fluid extraction of Croton crassifolius Geisel root: Chemical composition and anti-proliferative, autophagic, apoptosis-inducing, and related molecular effects on A549 tumour cells","volume":"61","author":"Liu","year":"2019","journal-title":"Phytomedicine"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Cevallos-Morillo, C., Cisneros-P\u00e9rez, P., Llive, R., Ricaurte, M., Reinoso, C., Meneses, M., Guam\u00e1n, M., and Palma-Cando, A. (2021). Croton lechleri Extracts as Green Corrosion Inhibitors of Admiralty Brass in Hydrochloric Acid. Molecules, 26.","DOI":"10.3390\/molecules26247417"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Gallego, R., Mart\u00ednez, M., Cifuentes, A., Ib\u00e1\u00f1ez, E., and Herrero, M. (2019). Development of a Green Downstream Process for the Valorization of Porphyridium cruentum Biomass. Molecules, 24.","DOI":"10.3390\/molecules24081564"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"101821","DOI":"10.1016\/j.jcou.2021.101821","article-title":"Valorization by compressed fluids of Arctium lappa seeds and roots as a sustainable source of valuable compounds","volume":"56","author":"Stefanov","year":"2022","journal-title":"J. CO2 Util."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.fitote.2014.03.012","article-title":"Two novel clerodane diterpenenes with NGF-potentiating activities from the twigs of Croton yanhuii","volume":"95","author":"Sun","year":"2014","journal-title":"Fitoterapia"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.phytol.2017.10.007","article-title":"Cytotoxic abietane-type diterpenoids from twigs and leaves of Croton laevigatus","volume":"22","author":"Song","year":"2017","journal-title":"Phytochem. Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1016\/j.bmcl.2015.01.033","article-title":"New clerodane diterpenoids from the twigs and leaves of Croton euryphyllus","volume":"25","author":"Pan","year":"2015","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_23","first-page":"44","article-title":"Sustainable Harvesting of Dragon\u2019s Blood (Croton lechleri) in Peru","volume":"125","author":"King","year":"2020","journal-title":"Herb. Gram."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Antony, A., and Farid, M. (2022). Effect of temperatures on polyphenols during extraction. Appl. Sci., 12.","DOI":"10.3390\/app12042107"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1021\/jf025878j","article-title":"Subcritical Water Extraction of Antioxidant Compounds from Rosemary Plants","volume":"51","author":"Cavero","year":"2003","journal-title":"J. Agric. Food Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1016\/j.biotechadv.2014.04.006","article-title":"Advances in the biotechnological glycosylation of valuable flavonoids","volume":"32","author":"Xiao","year":"2014","journal-title":"Biotechnol. Adv."},{"key":"ref_27","first-page":"6792069","article-title":"Environmentally Friendly Methods for Flavonoid Extraction from Plant Material: Impact of Their Operating Conditions on Yield and Antioxidant Properties","volume":"2020","year":"2020","journal-title":"Sci. World J."},{"key":"ref_28","first-page":"353","article-title":"Extraction of flavonoids from Buchanania lanzan spreng. seeds by supercritical fluid extraction and determination of their antioxidantactivity","volume":"8","author":"Vyavaharkar","year":"2016","journal-title":"Int. J. Pharm. Pharm. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4016","DOI":"10.1007\/s13197-019-03870-y","article-title":"Effects of high pressure extraction on the extraction yield, phenolic compounds, antioxidant and anti-tyrosinase activity of Djulis hull","volume":"56","author":"Huang","year":"2019","journal-title":"J. Food. Sci. Technol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/j.jep.2012.01.009","article-title":"Antitumor effect of Croton lechleri Mull. Arg. (Euphorbiaceae)","volume":"140","year":"2012","journal-title":"J. Ethnopharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1528","DOI":"10.1177\/147323000903700531","article-title":"The wound healing process: An overview of the cellular and molecular mechanisms","volume":"37","author":"Velnar","year":"2009","journal-title":"J. Int. Med. Res."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Wiegand, C., Hipler, U., Elsner, P., and Tittelbach, J. (2021). Keratinocyte and Fibroblast Wound Healing In Vitro Is Repressed by Non-Optimal Conditions but the Reparative Potential Can Be Improved by Water-Filtered Infrared A. Biomedicines, 9.","DOI":"10.3390\/biomedicines9121802"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e18761429260980","DOI":"10.2174\/0118761429260980231005105929","article-title":"RBM3 Accelerates Wound Healing of Skin in Diabetes through ERK1\/2 Signaling","volume":"17","author":"Feng","year":"2024","journal-title":"Curr. Mol. Pharmacol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1016\/j.ejcb.2007.03.009","article-title":"Cathepsin B is essential for regeneration of scratch-wounded normal human epidermal keratinocytes","volume":"86","author":"Buttigieg","year":"2007","journal-title":"Eur. J. Cell. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/S0031-9422(99)00513-0","article-title":"Importance of a pyrogallol-type structure in catechin compounds for apoptosis-inducing activity","volume":"53","author":"Saeki","year":"2000","journal-title":"Phytochemistry"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"12786","DOI":"10.1111\/dth.12786","article-title":"Review of future insights of Dragon\u2019s Blood in dermatology","volume":"32","author":"Pona","year":"2018","journal-title":"Dermatol. Ther."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1055\/s-2006-959567","article-title":"Studies on the anti-tumour, anti-bacterial, and wound-healing properties of dragon\u2019s blood","volume":"60","author":"Chen","year":"1994","journal-title":"Planta Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1078\/094471103321659843","article-title":"Evaluation of the mutagenic, antimutagenic and antiproliferative potential of Croton lechleri (Muell. Arg.) latex","volume":"10","author":"Rossi","year":"2003","journal-title":"Phytomedicine"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1248\/cpb.39.1041","article-title":"Cytotoxic Substance from Sangre de Grado","volume":"9","author":"Itokawa","year":"1991","journal-title":"Chem. Pharm. Bull."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"927","DOI":"10.2144\/000112812","article-title":"Detection of S-phase Cell Cycle Progression using 5-ethynyl-2\u2032-deoxyuridine Incorporation with Click Chemistry, an Alternative to using 5-bromo-2\u2032-deoxyuridine Antibodies","volume":"44","author":"Buck","year":"2008","journal-title":"BioTechniques"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Abate, M., Citro, M., Pisanti, S., Caputo, M., and Martinelli, R. (2021). Keratinocytes Migration Promotion, Proliferation Induction, and Free Radical Injury Prevention by 3-Hydroxytirosol. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22052438"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Yin, J., Kim, H., Hwang, I., Kim, D., and Lee, M. (2019). Anti-Inflammatory Effects of Phenolic Compounds Isolated from Quercus mongolica Fisch. ex Ledeb. on UVB-Irradiated Human Skin Cells. Molecules, 24.","DOI":"10.3390\/molecules24173094"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1016\/j.phymed.2013.10.006","article-title":"Flavonoids promoting HaCaT migration: I. Hologram quantitative structure\u2013activity relationships","volume":"21","author":"Cho","year":"2014","journal-title":"Phytomedicine"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.1021\/acs.jnatprod.2c00720","article-title":"Digital Database of Absorption Spectra of Diverse Flavonoids Enables Structural Comparisons and Quantitative Evaluations","volume":"86","author":"Taniguchi","year":"2023","journal-title":"J. Nat. Prod."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Alldritt, I., Whitham-Agut, B., Sipin, M., Studholme, J., Trentacoste, A., Tripp, J., Cappai, M., Ditchfield, P., Devi\u00e8se, T., and Hedges, R. (2019). Metabolomics reveals diet-derived plant polyphenols accumulate in physiological bone. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-44390-1"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.canlet.2006.01.027","article-title":"Green tea polyphenols and its constituent epigallocatechin gallate inhibits proliferation of human breast cancer cells in vitro and in vivo","volume":"245","author":"Thangapazham","year":"2007","journal-title":"Cancer Lett."},{"key":"ref_47","first-page":"611","article-title":"Inhibition of growth and induction of apoptosis in human cancer cell lines by tea polyphenols","volume":"19","author":"Yang","year":"1998","journal-title":"J. Leukoc. Biol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/j.fct.2017.04.021","article-title":"Galloylation of polyphenols alters their biological activity","volume":"105","author":"Karas","year":"2017","journal-title":"Food. Chem. Toxicol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1016\/j.jnutbio.2007.10.004","article-title":"Cytotoxic effects of digalloyl dimer procyanidins in human cancer cell lines","volume":"19","author":"Romanczyk","year":"2008","journal-title":"J. Nutr. Biochem."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Ho, K., Roy, A., Foote, S., Vo, P., Lall, N., and Lin, C. (2020). Profiling Anticancer and Antioxidant Activities of Phenolic Compounds Present in Black Walnuts (Juglans nigra) Using a High-Throughput Screening Approach. Molecules, 25.","DOI":"10.3390\/molecules25194516"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Qi, W., Qi, W., Xiong, D., and Long, M. (2022). Quercetin: Its Antioxidant Mechanism, Antibacterial Properties and Potential Application in Prevention and Control of Toxipathy. Molecules, 27.","DOI":"10.3390\/molecules27196545"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2033","DOI":"10.1016\/0031-9422(91)85063-6","article-title":"Polyphenolic compounds from Croton lechleri","volume":"30","author":"Cai","year":"1991","journal-title":"Phytochemistry"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/S0944-7113(11)80026-7","article-title":"SP-303, an antiviral oligomeric proanthocyanidin from the latex of Croton lechleri (Sangre de Drago)","volume":"1","author":"Ubillas","year":"1994","journal-title":"Phytomedicine"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Tsvetanova, F., Boyadzhieva, S., Coelho, J., Yankov, D., and Stateva, R. (2024). Sustainable transformation of two algal spe-cies of different genera to high-value chemicals and bioproducts. Molecules, 29.","DOI":"10.3390\/molecules29010156"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Boyadzhieva, S., Coelho, J., Errico, M., Reynel-Avilla, H., Yankov, D., Bonilla-Petriciolet, A., and Stateva, R. (2022). Assessment of Gnaphalium viscosum (Kunth) Valorization Prospects: Sustainable Recovery of Antioxidants by Different Techniques. Antioxidants, 11.","DOI":"10.3390\/antiox11122495"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"144","DOI":"10.5344\/ajev.1965.16.3.144","article-title":"Colorimetry of Total Phenolics with Phosphomolybdic-Phosphotungstic Acid Reagents","volume":"16","author":"Singleton","year":"1965","journal-title":"Am. J. Enol. Viticult."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S0023-6438(95)80008-5","article-title":"Use of a free radical method to evaluate antioxidant activity","volume":"28","author":"Cuvelier","year":"1995","journal-title":"LWT-Food Sci. Technol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"5032","DOI":"10.1021\/jf049571r","article-title":"Antioxidant activity of a flavonoid-rich extract of Hypericum perforatum L. in vitro","volume":"52","author":"Zou","year":"2004","journal-title":"J. Agric. Food Chem."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1007\/978-1-61779-080-5_20","article-title":"Cell Sensitivity Assays: The MTT Assay","volume":"731","author":"Kaspers","year":"2011","journal-title":"Methods Mol. Biol."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Stirling, D., Swain-Bowden, M., Lucas, A., Carpenter, A., Cimini, B., and Goodman, A. (2021). CellProfiler 4: Improvements in speed, utility and usability. BMC Bioinform., 22.","DOI":"10.1186\/s12859-021-04344-9"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.chroma.2013.12.070","article-title":"Isolation of high-purity anthocyanin mixtures and monomers from blueberries using combined chromatographic techniques","volume":"1327","author":"Wang","year":"2014","journal-title":"J. Chromatogr. A"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/17\/4161\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:47:31Z","timestamp":1760111251000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/17\/4161"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,2]]},"references-count":61,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2024,9]]}},"alternative-id":["molecules29174161"],"URL":"https:\/\/doi.org\/10.3390\/molecules29174161","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,9,2]]}}}