利用高效液相色谱法从菲律宾医蛭中分离纯化新型水蛭素

Isolation and purification of novel hirudins from the leech Hirudinaria manillensis by high-performance liquid chromatography.

作者信息

Steiner V, Knecht R, Gruetter M, Raschdorf F, Gassmann E, Maschler R

机构信息

Pharmaceuticals Research Laboratories, Ciba-Geigy Ltd., Basel, Switzerland.

出版信息

J Chromatogr. 1990 Sep 14;530(2):273-82. doi: 10.1016/s0378-4347(00)82331-2.

DOI:10.1016/s0378-4347(00)82331-2

PMID:2079503

Abstract

The isolation and purification of novel hirudins from a crude extract of the leech Hirudinaria manillensis and their analytical characterization are reported. Initial purification by gel permeation chromatography on Sephadex G50 and anion-exchange chromatography on Q Sepharose fast-flow removed most contaminants and yielded a highly active extract. Two isohirudins (designated hirudin P6 and P18) were isolated and purified by successive reversed-phase high-performance liquid chromatography on silica-based stationary phases and anion-exchange chromatography on Mono Q. The final products were characterized by reversed-phase high-performance liquid chromatography, 252Cf plasma desorption time-of-flight mass spectrometry and capillary zone electrophoresis. The molecular masses determined by 252Cf plasma desorption mass spectrometry were 7416 dalton for hirudin P6 and 7199 dalton for hirudin P18.

摘要

报道了从马尼拉医蛭粗提物中分离纯化新型水蛭素及其分析表征。通过在Sephadex G50上进行凝胶渗透色谱和在Q Sepharose fast-flow上进行阴离子交换色谱进行初步纯化，去除了大部分污染物，得到了一种高活性提取物。通过在硅胶基固定相上连续进行反相高效液相色谱和在Mono Q上进行阴离子交换色谱，分离并纯化了两种异水蛭素（分别命名为水蛭素P6和P18）。通过反相高效液相色谱、252Cf等离子体解吸飞行时间质谱和毛细管区带电泳对最终产物进行了表征。通过252Cf等离子体解吸质谱测定，水蛭素P6的分子量为7416道尔顿，水蛭素P18的分子量为7199道尔顿。