A rapid method for isolation and purification of an anticoagulant from Whitmania pigra |
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Authors: | Zhong Shan Cui Zheng Sakura Naoki Wang Dong Li Jianlin Zhai Yan |
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Institution: | Department of Pharmacognosy, School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, People's Republic of China. |
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Abstract: | Whitmania pigra is common in China and has been used as a traditional Chinese anticoagulant medicine for years, but its effective components are unknown to scientists. In this article we report a rapid method for isolation and purification of an anticoagulant from W. pigra for the first time. An acetone-water extract of W. pigra was subjected to anion-exchange chromatography on a Sephadex DEAE A-50 column, and gel permeation chromatography on Sephadex G-25 and Sephadex LH-20 columns successively, which afforded a fraction with potent anticoagulant activity. An anticoagulant was isolated and purified from this fraction by reversed-phase high-performance liquid chromatography (RP-HPLC). It was identified as a single pure substance by RP-HPLC and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). This component was named whitmanin and its molecular weight was determined as 8608 Da by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). |
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Keywords: | leech Whitmania pigra whitmanin HPLC MALDI‐TOF‐MS Sephadex DEAE A‐50 Sephadex G‐25 Sephadex LH‐20 SDS‐PAGE |
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