Studies on anticoagulant constituents in dried Whitmania pigra / 中国中药杂志

ABSTRACT

<p><b>OBJECTIVE</b>To study the anticoagulant constituents in dried leech (Whitmania pigra Whitman).</p><p><b>METHOD</b>The plasma recalcification time (PRT) as index, the constituents with anticoagulant activity were isolated and purified by anion-exchange chromatography on Sephadex DEAE A-50, gel permeation chromatography on Sephadex G-25 and Sephadex LH-20 columns, and then reversed phase high-performance liquid chromatography successively.</p><p><b>RESULT</b>Three anticoagulant polypeptides were isolated and purified. Compounds 1 and 2 can be translated each other in natural conditions, and their molecular weights are 7100 and 5531, respectively. Compound 3 was identified as a pure polypeptide by HPLC and SDS-PAGE, and its molecular weight was determined as 8 608 by MALDI-TOF-MS. The amino acid composition of compound 3 was also determined.</p><p><b>CONCLUSION</b>Compound 3 was inferred to be a novel anticoagulant, and named whitmanin.</p>