Zhong Shan, Cui Zheng, Sakura Naoki, Wang Dong, Li Jianlin, Zhai Yan
Department of Pharmacognosy, School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, People's Republic of China.
Biomed Chromatogr. 2007 May;21(5):439-45. doi: 10.1002/bmc.739.
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).
宽体金线蛭在中国很常见，多年来一直被用作传统的中国抗凝血药物，但科学家们对其有效成分并不了解。在本文中，我们首次报道了一种从宽体金线蛭中分离和纯化抗凝血剂的快速方法。宽体金线蛭的丙酮 - 水提取物先后在Sephadex DEAE A - 50柱上进行阴离子交换色谱，以及在Sephadex G - 25和Sephadex LH - 20柱上进行凝胶渗透色谱，得到了具有强抗凝血活性的级分。通过反相高效液相色谱（RP - HPLC）从该级分中分离和纯化出一种抗凝血剂。通过RP - HPLC和十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS - PAGE）将其鉴定为单一纯物质。该成分被命名为水蛭素，通过基质辅助激光解吸电离/飞行时间质谱（MALDI - TOF - MS）测定其分子量为8608 Da。
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