{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T20:31:38Z","timestamp":1790713898123,"version":"4.1.0"},"reference-count":33,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,3,29]],"date-time":"2022-03-29T00:00:00Z","timestamp":1648512000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["2018M3A9F3081538"],"award-info":[{"award-number":["2018M3A9F3081538"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>The flower buds of Daphne genkwa have been reported as a potent resource associated with anti-angiogenic, anti-tumor, anti-rheumatoid arthritis activities, as well as immunoregulation. This paper aimed to establish an optimal extraction method for flavonoids, as active phytochemicals, and to conduct a comparative analysis by profiling the different blooming stages. Optimized shaking extraction conditions from the design of experiments (DoE), such as minutely mixture design, 23 full factorial design, and polynomial regression analysis, involved an agitation speed of 150 rpm and temperature of 65 \u00b0C for 12 h in 56% (v\/v) acetone solvent. After, a comparative analysis was performed on three blooming stages, juvenile bud, mature purple bud, and complete flowering, by ultra-high-performance liquid chromatography-photodiode array-mass spectrometry (UHPLC-PDA-MS). Most flavonoids increased during bud growth and then decreased when the bud opened for blooming. In particular, apigenin 7-O-glucuronide, genkwanin 5-O-primeveroside, and genkwanin strikingly showcased this pattern. Furthermore, the raw spectrometric dataset was subjected to orthogonal projection to latent structures discriminant analysis (OPLS-DA) to find significant differences in the flavonoids from the juvenile bud, mature purple bud, and complete flowering. In conclusion, the present study facilitates an understanding of flavonoid change at different blooming stages and provides a momentous reference in the research of D. genkwa.<\/jats:p>","DOI":"10.3390\/plants11070925","type":"journal-article","created":{"date-parts":[[2022,3,29]],"date-time":"2022-03-29T21:41:30Z","timestamp":1648590090000},"page":"925","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Design of Experiments-Based Optimization of Flavonoids Extraction from Daphne genkwa Flower Buds and Flavonoids Contents at Different Blooming Stages"],"prefix":"10.3390","volume":"11","author":[{"given":"Min-Kyoung","family":"Kim","sequence":"first","affiliation":[{"name":"Division of Pharmacognosy, College of Pharmacy, Kyung Hee University, Seoul 02447, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7072-2602","authenticated-orcid":false,"given":"Geonha","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Life and Nanopharmaceutical Sciences, Graduate School, Kyung Hee University, Seoul 02447, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yura","family":"Ji","sequence":"additional","affiliation":[{"name":"Department of Life and Nanopharmaceutical Sciences, Graduate School, Kyung Hee University, Seoul 02447, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun-Gyo","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Pharmaceutical Sciences, Graduate School, Kyung Hee University, Seoul 02447, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minsik","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Pharmaceutical Sciences, Graduate School, Kyung Hee University, Seoul 02447, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seung-Hyeon","family":"Go","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Pharmaceutical Sciences, Graduate School, Kyung Hee University, Seoul 02447, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miwon","family":"Son","sequence":"additional","affiliation":[{"name":"Central Research Center, Mtherapharma Co., Seoul 07793, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5865-9228","authenticated-orcid":false,"given":"Young-Pyo","family":"Jang","sequence":"additional","affiliation":[{"name":"Department of Life and Nanopharmaceutical Sciences, Graduate School, Kyung Hee University, Seoul 02447, Korea"},{"name":"Department of Biomedical and Pharmaceutical Sciences, Graduate School, Kyung Hee University, Seoul 02447, Korea"},{"name":"Department of Oriental Pharmaceutical Science, College of Pharmacy, Kyung Hee University, Seoul 02447, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1007\/s11101-019-09625-z","article-title":"Botanical ingredient identification and quality assessment: Strengths and limitations of analytical techniques","volume":"19","author":"Upton","year":"2020","journal-title":"Phytochem. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1002\/cbdv.201000117","article-title":"Phytochemical and biological studies of the plants from the genus Daphne","volume":"8","author":"Xu","year":"2011","journal-title":"Chem. Biodivers."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"104055","DOI":"10.1016\/j.bse.2020.104055","article-title":"Chemical constituents from the flower buds of Daphne genkwa (Thymelaeaceae)","volume":"91","author":"Hou","year":"2020","journal-title":"Biochem. Syst. Ecol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"185","DOI":"10.2174\/1389557024605492","article-title":"Daphnane-type diterpene orthoesters and their biological activities","volume":"2","author":"He","year":"2002","journal-title":"Mini Rev. Med. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1248\/cpb.58.234","article-title":"Daphnane diterpene esters with anti-proliferative activities against human lung cancer cells from Daphne genkwa","volume":"58","author":"Hong","year":"2010","journal-title":"Chem. Pharm. Bull."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2500","DOI":"10.1016\/j.bmcl.2013.03.025","article-title":"Daphnane-type diterpenes with inhibitory activities against human cancer cell lines from Daphne genkwa","volume":"23","author":"Li","year":"2013","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1604","DOI":"10.1021\/acs.jnatprod.6b00110","article-title":"Daphnane diterpenes from Daphne genkwa activate Nurr1 and have a neuroprotective effect in an animal model of Parkinson\u2019s disease","volume":"79","author":"Han","year":"2016","journal-title":"J. Nat. Prod."},{"key":"ref_8","first-page":"S1","article-title":"Nurr1-Activating Daphnane Diterpenes from Daphne genkwa and its Neuroprotective Effect in an Animal Model of Parkinson\u2019s Disease","volume":"4","author":"Han","year":"2017","journal-title":"Planta Med. Int. Open"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2205","DOI":"10.1248\/bpb.b17-00641","article-title":"Daphnane and phorbol diterpenes, anti-neuroinflammatory compounds with Nurr1 activation from the roots and stems of Daphne genkwa","volume":"40","author":"Han","year":"2017","journal-title":"Biol. Pharm. Bull."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"104540","DOI":"10.1016\/j.fitote.2020.104540","article-title":"The genus Daphne: A review of its traditional uses, phytochemistry and pharmacology","volume":"143","author":"Moshiashvili","year":"2020","journal-title":"Fitoterapia"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"113683","DOI":"10.1016\/j.jep.2020.113683","article-title":"Comprehensive evaluation on anti-inflammatory and anti-angiogenic activities in vitro of fourteen flavonoids from Daphne Genkwa based on the combination of efficacy coefficient method and principal component analysis","volume":"268","author":"Zhou","year":"2021","journal-title":"J. Ethnopharmacol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"106384","DOI":"10.1016\/j.intimp.2020.106384","article-title":"Anti-rheumatoid arthritis effects of flavonoids from Daphne genkwa","volume":"83","author":"Sun","year":"2020","journal-title":"Int. Immunopharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1002\/ptr.5540","article-title":"Antitumor activity of total flavonoids from Daphne genkwa in colorectal cancer","volume":"30","author":"Du","year":"2016","journal-title":"Phytother. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.intimp.2006.07.010","article-title":"Total flavonoids of Daphne genkwa root significantly inhibit the growth and metastasis of Lewis lung carcinoma in C57BL6 mice","volume":"7","author":"Zheng","year":"2007","journal-title":"Int. Immunopharmacol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"13414","DOI":"10.1038\/s41598-018-31831-6","article-title":"Transcriptome analysis of differentially expressed unigenes involved in flavonoid biosynthesis during flower development of Chrysanthemum morifolium \u2018Chuju\u2019","volume":"8","author":"Yue","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1007\/s12298-021-00945-z","article-title":"Metabolic profiling and antioxidant activity during flower development in Agastache rugosa","volume":"27","author":"Yeo","year":"2021","journal-title":"Physiol. Mol. Biol. Plants"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Park, C.H., Yeo, H.J., Kim, Y.J., Nguyen, B.V., Park, Y.E., Sathasivam, R., Kim, J.K., and Park, S.U. (2021). Profiles of Secondary Metabolites (Phenolic Acids, Carotenoids, Anthocyanins, and Galantamine) and Primary Metabolites (Carbohydrates, Amino Acids, and Organic Acids) during Flower Development in Lycoris radiata. Biomolecules, 11.","DOI":"10.3390\/biom11020248"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"207","DOI":"10.11110\/kjpt.2017.47.3.207","article-title":"A comparative morphological study of Thymelaeaceae in Korea","volume":"47","author":"Lee","year":"2017","journal-title":"Korean J. Plant Taxon."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"11936","DOI":"10.1038\/s41598-021-91341-w","article-title":"Analytical quality by design methodology for botanical raw material analysis: A case study of flavonoids in Genkwa Flos","volume":"11","author":"Kim","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1016\/j.matpr.2020.06.466","article-title":"Optimization of process parameters using response surface methodology: A review","volume":"37","author":"Chelladurai","year":"2021","journal-title":"Mater. Today Proc."},{"key":"ref_21","first-page":"98","article-title":"Thermodynamic characteristics of solvents: A review","volume":"3","author":"Shafique","year":"2013","journal-title":"Res. J. Chem. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.foodchem.2017.11.114","article-title":"Multivariate optimization techniques in food analysis\u2014A review","volume":"273","author":"Ferreira","year":"2019","journal-title":"Food Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e14357","DOI":"10.1111\/jfpp.14357","article-title":"Bioactive properties and phenolic compounds in bud, sprout, and fruit of Capparis spp. plants","volume":"44","author":"Ghafoor","year":"2020","journal-title":"J. Food Process. Preserv."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1016\/j.jfoodeng.2013.01.014","article-title":"Techniques for extraction of bioactive compounds from plant materials: A review","volume":"117","author":"Azmir","year":"2013","journal-title":"J. Food Eng."},{"key":"ref_25","first-page":"2167-0412","article-title":"A review on the extraction methods use in medicinal plants, principle, strength and limitation","volume":"4","author":"Azwanida","year":"2015","journal-title":"Med. Aromat. Plants"},{"key":"ref_26","unstructured":"Antony, J. (2014). Design of Experiments for Engineers and Scientists, Elsevier."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Altemimi, A., Lakhssassi, N., Baharlouei, A., Watson, D.G., and Lightfoot, D.A. (2017). Phytochemicals: Extraction, Isolation, and Identification of Bioactive Compounds from Plant Extracts. Plants, 6.","DOI":"10.3390\/plants6040042"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2167","DOI":"10.3390\/molecules14062167","article-title":"Effect of extraction solvent\/technique on the antioxidant activity of selected medicinal plant extracts","volume":"14","author":"Sultana","year":"2009","journal-title":"Molecules"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1038\/s41592-019-0335-9","article-title":"Two-level factorial experiments","volume":"16","author":"Smucker","year":"2019","journal-title":"Nat. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.foodchem.2014.08.080","article-title":"Optimisation of ultrasound-assisted extraction conditions for maximal recovery of active monacolins and removal of toxic citrinin from red yeast rice by a full factorial design coupled with response surface methodology","volume":"170","author":"Zhou","year":"2015","journal-title":"Food Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1002\/aic.690030308","article-title":"Surface tension of acetone-water solutions up to their normal boiling points","volume":"3","author":"Howard","year":"1957","journal-title":"AIChE J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1039\/C4MB00414K","article-title":"PLS\/OPLS models in metabolomics: The impact of permutation of dataset rows on the K-fold cross-validation quality parameters","volume":"11","author":"Triba","year":"2015","journal-title":"Mol. BioSyst."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Lee, S.L. (2019). A Totality-of-evidence approach to ensuring therapeutic consistency of naturally derived complex mixtures. The Science and Regulations of Naturally Derived Complex Drugs, Springer.","DOI":"10.1007\/978-3-030-11751-1_15"}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/11\/7\/925\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:46:12Z","timestamp":1760136372000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/11\/7\/925"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,29]]},"references-count":33,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,4]]}},"alternative-id":["plants11070925"],"URL":"https:\/\/doi.org\/10.3390\/plants11070925","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,29]]}}}