{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T21:32:04Z","timestamp":1761514324281,"version":"build-2065373602"},"reference-count":28,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2019,2,25]],"date-time":"2019-02-25T00:00:00Z","timestamp":1551052800000},"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":["31670358","31870336"],"award-info":[{"award-number":["31670358","31870336"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007847","name":"Natural Science Foundation of Jilin Province","doi-asserted-by":"publisher","award":["20180414028GH","201807414028GH"],"award-info":[{"award-number":["20180414028GH","201807414028GH"]}],"id":[{"id":"10.13039\/100007847","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010211","name":"Education Department of Jilin Province","doi-asserted-by":"publisher","award":["JJKH20181171KJ"],"award-info":[{"award-number":["JJKH20181171KJ"]}],"id":[{"id":"10.13039\/501100010211","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Separation Science"],"published-print":{"date-parts":[[2019,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Nonsteroidal anti\u2010inflammatory drugs reportedly reduce the risk of developing cancer. One mechanism by which they reduce carcinogenesis involves the inhibition of the activity of cyclooxygenase\u20102, an enzyme that is overexpressed in various cancer tissues. Its overexpression increases cell proliferation and inhibits apoptosis. However, selected cyclooxygenase\u20102 inhibitors can also act through cyclooxygenase\u2010independent mechanisms. In this study, using ultrafiltration, enzyme\u2010immobilized magnetic beads, high\u2010performance liquid chromatography, and electrospray\u2010ionization mass spectrometry, several isoflavonoids in <jats:italic>Trifolium pratense L<\/jats:italic>. extracts were screened and identified. Semi\u2010preparative high\u2010performance liquid chromatography and high\u2010speed counter\u2010current chromatography were then applied to separate the active constituents. Using these methods, seven major compounds were identified in <jats:italic>Trifolium pratense L<\/jats:italic>. As cyclooxygenase\u20102 inhibitors: rothindin, ononin, daidzein, trifoside, pseudobaptigenin, formononetin, and biochanin A, which were then isolated with &gt;92% purity. This is the first report of the presence of potent cyclooxygenase\u20102 inhibitors in <jats:italic>Trifolium pratense L<\/jats:italic>. extracts. The results of this study demonstrate that the systematic isolation of bioactive components from <jats:italic>Trifolium pratense L<\/jats:italic>., by using ultrafiltration, enzyme\u2010immobilized magnetic beads, semi\u2010preparative high\u2010performance liquid chromatography, and high\u2010speed counter\u2010current chromatography, represents a feasible and efficient technique that could be extended for the identification and isolation of other enzyme inhibitors.<\/jats:p>","DOI":"10.1002\/jssc.201800986","type":"journal-article","created":{"date-parts":[[2019,1,9]],"date-time":"2019-01-09T00:20:18Z","timestamp":1546993218000},"page":"1133-1143","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Screening and isolation of cyclooxygenase\u20102 inhibitors from <i>Trifolium pratense L<\/i>. via ultrafiltration, enzyme\u2010immobilized magnetic beads, semi\u2010preparative high\u2010performance liquid chromatography and high\u2010speed counter\u2010current chromatography"],"prefix":"10.1002","volume":"42","author":[{"given":"Wanchao","family":"Hou","sequence":"first","affiliation":[{"name":"Central Laboratory Changchun Normal University  Erdao District Changchun P. 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