A rapid method for isolation and purification of an anticoagulant from Whitmania pigra

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). Copyright © 2007 John Wiley & Sons, Ltd.