American Society of Hirudotherapy

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

Biochemistry/pharmacology published in Biomed Chromatogr (2007)

Last Updated: June 18, 2026Reviewed by: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: Preclinical (animal)Salivary PharmacologyZhong S et al. · Biomedical chromatography : BMC, 2007

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).

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

Publication typeJournal Article
Indexed MeSH termsAnimalsAnnelidaAnticoagulantsChromatography, GelChromatography, High Pressure LiquidChromatography, Ion ExchangeElectrophoresis, Polyacrylamide GelSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Summary

First isolation of 'whitmanin', an 8,608 Da anticoagulant protein purified by anion-exchange, gel-permeation, and reversed-phase HPLC from Whitmania pigra acetone-water extract.

Why This Matters for Hirudotherapy

This study reports a rapid method for isolating and purifying an anticoagulant from *Whitmania pigra*, a species used in traditional Chinese medicine. Using various chromatography techniques, researchers obtained a fraction with potent anticoagulant activity, from which they successfully purified a single substance named whitmanin. The compound was identified as pure, and its molecular weight was determined to be 8608 Da using mass spectrometry. While this research identifies a specific anticoagulant compound from a leech species, the abstract's scope is strictly limited to extraction and physicochemical characterization, providing no data on in vivo models, clinical efficacy, or its use in live hirudotherapy.

Citation

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

Zhong S et al. · Biomedical chromatography : BMC, 2007

DOI

Added to ASH library: May 26, 2026 · Site last updated: June 18, 2026

This website provides educational information and does not constitute medical advice, diagnosis, or treatment recommendations. Medicinal leech therapy carries clinically meaningful risks and should be performed only by qualified clinicians under institutionally approved protocols. FDA 510(k) clearance for medicinal leeches is limited to specific indications; investigational and off-label discussions are labeled accordingly. For patient-specific guidance, consult a qualified healthcare provider.