{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T13:48:27Z","timestamp":1783604907028,"version":"3.55.0"},"reference-count":25,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T00:00:00Z","timestamp":1753401600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82104358"],"award-info":[{"award-number":["82104358"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["Number 82104358"],"award-info":[{"award-number":["Number 82104358"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003787","name":"Hebei Provincial Natural Science Foundation of China","doi-asserted-by":"publisher","award":["Number H2021423066"],"award-info":[{"award-number":["Number H2021423066"]}],"id":[{"id":"10.13039\/501100003787","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Phytochemical Analysis"],"published-print":{"date-parts":[[2025,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>Postmenopausal osteoporosis, a skeletal disorder induced by estrogen deficiency, is commonly addressed in clinical practice through the use of Cuscutae semen and its compound formulations, owing to their significant therapeutic efficacy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>This study sought to systematically identify bioactive compounds present in Cuscutae semen that interact with estrogen receptor \u03b2 (ESR2) using Affinity Ultrafiltration combined with Ultra\u2010Performance Liquid Chromatography\u2010Quadrupole\u2010Orbitrap Mass Spectrometry (UPLC\u2010QE\u2010Orbitrap\u2010MS) to find possible therapy options for PMOP.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The Cuscutae semen extract was fractionated using a C18 column to obtain fractions based on distinct polarity. Preliminary evaluation of anti\u2010osteoporotic activity was conducted in MC3T3\u2010E1 cells by assessing osteoprotegerin (OPG) expression. Affinity Ultrafiltration integrated with UPLC\u2010QE\u2010Orbitrap\u2010MS was utilized to screen for ligands binding to ESR2, followed by molecular docking to validate the interaction mechanisms. The osteogenic effects of the identified compounds were further confirmed through CCK\u20108 proliferation assays, OPG quantification, and alkaline phosphatase (ALP) activity analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The fraction of Cuscutae semen extract eluted with 100% methanol exhibited significant anti\u2010osteoporotic activity. Three flavonoids\u2014Astragalin, Isorhamnetin, and Quercitrin\u2014that interact with ESR2 were successfully identified. In\u00a0vitro validation demonstrated the efficacy of these compounds.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study presents a comprehensive strategy that integrates Affinity Ultrafiltration, UPLC\u2010QE\u2010Orbitrap\u2010MS, and bioactivity validation to efficiently identify ESR2\u2010targeted postmenopausal osteoporosis compounds in Cuscutae Semen. The findings offer both theoretical and empirical foundations for the development of innovative therapeutics for postmenopausal osteoporosis.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pca.70018","type":"journal-article","created":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T10:40:37Z","timestamp":1753440037000},"page":"2119-2128","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["High\u2010Efficiency Screening of Compounds Targeting Postmenopausal Osteoporosis in Cuscutae Semen Based on Affinity Ultrafiltration\u2010UPLC\u2010QE\u2010Orbitrap\u2010MS Platform"],"prefix":"10.1002","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-8635-3896","authenticated-orcid":false,"given":"Li","family":"Yishan","sequence":"first","affiliation":[{"name":"Jilin Ginseng Academy Changchun University of Chinese Medicine  Changchun China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Yawen","sequence":"additional","affiliation":[{"name":"Jilin Ginseng Academy Changchun University of Chinese Medicine  Changchun China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liu","family":"Wanjie","sequence":"additional","affiliation":[{"name":"Jilin Ginseng Academy Changchun University of Chinese Medicine  Changchun China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Kunping","sequence":"additional","affiliation":[{"name":"Jilin Ginseng Academy Changchun University of Chinese Medicine  Changchun China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0048-4020","authenticated-orcid":false,"given":"Feng","family":"Wei","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Quality Evaluation &amp; 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