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  • Title: Isolation and purification of novel hirudins from the leech Hirudinaria manillensis by high-performance liquid chromatography.
    Author: Steiner V, Knecht R, Gruetter M, Raschdorf F, Gassmann E, Maschler R.
    Journal: J Chromatogr; 1990 Sep 14; 530(2):273-82. PubMed ID: 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.
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