{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:51:37Z","timestamp":1786978297789,"version":"build-2736575974"},"reference-count":32,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2024,3,18]],"date-time":"2024-03-18T00:00:00Z","timestamp":1710720000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100019081","name":"Science and Technology Program of Hunan Province","doi-asserted-by":"publisher","award":["2021JJ30511"],"award-info":[{"award-number":["2021JJ30511"]}],"id":[{"id":"10.13039\/501100019081","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":[[2024,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Polygonum amplexicaule<\/jats:italic>\n                      D. Don var.\n                      <jats:italic>sinense<\/jats:italic>\n                      Forb (PAF), a medicinal plant, has the effect of promoting blood circulation and removing blood stasis. However, the active compounds and targets of its anticoagulant effect are still unclear.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>This study aims to establish an effective reversely thrombin\u2010targeted screening method for anticoagulant active components in PAF by affinity ultrafiltration (AUF) coupled with ultrahigh\u2010performance liquid chromatography\u2013quadrupole time\u2010of\u2010flight mass spectroscopy (UPLC\u2010Q\u2010TOF\u2010MS).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Different polar parts of PAF were screened for potential thrombin ligands by AUF\u2010HPLC and identified by UPLC\u2010Q\u2010TOF\u2010MS. After studying the affinity between ligands and thrombin by molecular docking, the antithrombotic activity of ligands was detected in vivo by zebrafish thrombus model, and in vitro by chromogenic substrate method. The mechanism of such ligands on thrombin was further studied by coagulation factor assay.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Eleven potential thrombin ligands from PAF were screened by the AUF\u2010UPLC\u2010Q\u2010TOF\u2010MS method, and two compounds (butyl gallate and \u03b2\u2010sitosterol) with significant anticoagulant activity were discovered via in vitro and in vivo activity testing.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>A method system based on AUF\u2010UPLC\u2010Q\u2010TOF\u2010MS, molecular docking and in vivo and in vitro experiments also provided a powerful tool for further exploration of anticoagulant active components in PAF.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/pca.3346","type":"journal-article","created":{"date-parts":[[2024,3,19]],"date-time":"2024-03-19T03:36:05Z","timestamp":1710819365000},"page":"1112-1122","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Thrombin\u2010targeted screening of anticoagulant active components from\n                    <i>Polygonum amplexicaule<\/i>\n                    D. Don var.\n                    <i>sinense<\/i>\n                    Forb by affinity ultrafiltration coupled with UPLC\u2010Q\u2010TOF\u2010MS"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-6094-7320","authenticated-orcid":false,"given":"Shiyi","family":"Huang","sequence":"first","affiliation":[{"name":"School of Pharmacy Hunan University of Chinese Medicine  Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangchang","family":"He","sequence":"additional","affiliation":[{"name":"School of Pharmacy Hunan University of Chinese Medicine  Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7361-0131","authenticated-orcid":false,"given":"Chencun","family":"Huang","sequence":"additional","affiliation":[{"name":"School of Pharmacy Hunan University of Chinese Medicine  Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weihe","family":"He","sequence":"additional","affiliation":[{"name":"School of Pharmacy Hunan University of Chinese Medicine  Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongqing","family":"Zhao","sequence":"additional","affiliation":[{"name":"School of Pharmacy Hunan University of Chinese Medicine  Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Dai","sequence":"additional","affiliation":[{"name":"School of Pharmacy Hunan University of Chinese Medicine  Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangming","family":"Xu","sequence":"additional","affiliation":[{"name":"School of Pharmacy Hunan University of Chinese Medicine  Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,3,18]]},"reference":[{"issue":"23","key":"e_1_2_6_2_1","first-page":"2932","article-title":"Research progress of anticoagulant drugs for prevention and treatment of venous thromboembolism in tumor patients","volume":"31","author":"Li P","year":"2020","journal-title":"China Pharm"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature06797"},{"issue":"14","key":"e_1_2_6_4_1","first-page":"256","article-title":"Antithrombotic effects of Polygonum amplexicaule decoction and the action of mechanism","volume":"19","author":"Zeng BY","year":"2013","journal-title":"Chin J Exp Tradit Med Formulae"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fct.2015.02.020"},{"issue":"2","key":"e_1_2_6_6_1","first-page":"431","article-title":"Evaluation of antioxidant potential and HPLC based identification of phenolics in Polygonum amplexicaule extract and its fractions","volume":"28","author":"Batool S","year":"2015","journal-title":"Pak J Pharm Sci"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2017.00228"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2022.115460"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/md21100502"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2016.09.021"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/wdev.312"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.115.306823"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13020\u2010020\u201000316\u2010y"},{"key":"e_1_2_6_14_1","volume-title":"The Zebrafish Book. 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