Gel chromatography of heparin

Area of Science:

• Biochemistry
• Pharmacology
• Chromatography

Background:

• Heparin is a complex, heterogeneous anticoagulant substance.
• Understanding heparin heterogeneity is crucial for effective purification and therapeutic application.

Purpose of the Study:

• To investigate the purification of heparin through column chromatography.
• To fractionate heparin using cross-linked dextran gels and an automated apparatus.

Main Methods:

• Column chromatography utilizing various diameters of bead form cross-linked dextran gels.
• Automated apparatus for heparin fractionation.
• Assessment of anticoagulant activity using recalcification time, partial thromboplastin time, thrombin time, and anti-Xa tests.
• Electrophoresis for metachromasia analysis.

Main Results:

• Sephadex G-50 demonstrated superior resolution, separating heparin into three distinct peaks.
• One isolated peak (51% of original material) exhibited potent anticoagulant activity.
• This active fraction showed strong metachromasia and significant in vivo anticoagulant effects.

Conclusions:

• Column chromatography with Sephadex G-50 is an effective method for heparin fractionation.
• A specific heparin fraction possesses potent anticoagulant properties.
• This technique holds promise for purifying heparin from tissue sources for therapeutic applications.