Add time:08/29/2019         Source:sciencedirect.com

Heparin and low molecular weight heparin have been used widely in clinical therapy as anticoagulants in cardiovascular disease and in hemodialysis. Crude heparin is usually prepared from porcine intestinal mucosa. Purified heparin is a mixture of polysaccharides consisting mainly of repeating GlcNS(6S)-IdoA2S disaccharides and other disaccharides with different GlcNAc/GlcNS±3S±6S-GlcA/IdoA±2S residues. Heparin injections are prepared by purification and then sterilization of ungraded heparin. Low molecular weight heparins are dominant heparin-based drugs used clinically, which are prepared by degrading heparin into smaller molecules. As a result, low molecular weight heparins share the same major disaccharides with heparin, but may have different reducing and non-reducing ends due to different preparation methods. In the current study, we analyzed the disaccharide composition of clinically used heparin and heparin-based drugs. Heparinase I, II and III were used to degrade all heparin and heparin-based drugs including heparin sodium injection, Enoxaparin sodium injection, Nadroparin calcium injection, Dalteparin sodium injection, and Fondaparinux sodium injection into disaccharides. All the degraded products were analyzed by strong anion exchange-high performance liquid chromatography (SAX-HPLC) equipped with an online UV-detector. Commercially available unsaturated disaccharide standards were then used for structural identification. Furthermore, unusual disaccharides present in Nadroparin, Dalteparin and Fondaparinux were confirmed by reversed-phase ion-pair HPLC coupled with mass spectrometry analysis. The developed method produced detailed structural information, which should be useful for quality control of heparin and heparin-based drugs.

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