Heparinase Digestion-based Disaccharide Analysis of Clinical Heparin and Heparinoids Drug
10.11895/j.issn.0253-3820.150032
- VernacularTitle:临床肝素类药物酶解分析二糖组成
- Author:
Zhangrun HAN
;
Xinhui XING
;
Guangli YU
;
Yangyang ZENG
;
Lijuan ZHANG
- Publication Type:Journal Article
- Keywords:
Heparin;
Low molecular weight heparin;
Heparinase;
Disaccharide analysis;
Mass spectrometry
- From:
Chinese Journal of Analytical Chemistry
2015;(7):964-970
- CountryChina
- Language:Chinese
-
Abstract:
Heparin and low molecular weight heparin have been widely used 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 drugs prepared from heparin active pharmaceutical ingredient ( API ) that is prepared from crude heparin. Low molecular weight heparins are dominant heparin-based drugs used clinically, which are prepared by degrading heparin into smaller sizes. As a result, low molecular weight heparins are sharing the same major disaccharides but have different reducing and non-reducing ends. In current study, we focused on the disaccharide compositional analysis of clinically used heparin and heparin-based drugs. HeparinaseⅠ,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, Fondaparinux sodium into disaccharides. All the degraded products were analyzed by strong anion high perforance liquid chromatography ( SAX-HPLC) coupled with an UV-detector. Commercially available unsaturated disaccharide standards were then used for structral identification. Furthermore, unusual disaccharides present in Nadroparin, Dalteparin, and Fondaparinux were confirmed by reversed-phase ion pair HPLC coupled with mass spectrometry. The developed method produced detailed structural information, which should be useful for quality control of heparin and heparin-based drugs.
