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Characterization of Danaparoid Complex Extractive Drug by an Orthogonal Analytical Approach
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Characterization of Danaparoid Complex Extractive Drug by an Orthogonal Analytical Approach
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Journal Article
Characterization of Danaparoid Complex Extractive Drug by an Orthogonal Analytical Approach
2017
Overview
Danaparoid sodium salt, is the active component of ORGARAN, an anticoagulant and antithrombotic drug constituted of three glycosaminoglycans (GAGs) obtained from porcine intestinal mucosa extracts. Heparan sulfate is the major component, dermatan sulfate and chondroitin sulfate being the minor ones. Currently dermatan sulfate and chondroitin sulfate are quantified by UV detection of their unsaturated disaccharides obtained by enzymatic depolymerization. Due to the complexity of danaparoid biopolymers and the presence of shared components, an orthogonal approach has been applied using more advanced tools and methods. To integrate the analytical profile, 2D heteronuclear single quantum coherence (HSQC) NMR spectroscopy was applied and found effective to identify and quantify GAG component signals as well as those of some process signatures of danaparoid active pharmaceutical ingredient (API) batches. Analyses of components of both API samples and size separated fractions proceeded through the determination and distribution of the molecular weight (Mw) by high performance size exclusion chromatographic triple detector array (HP-SEC-TDA), chain mapping by LC/MS, and mono- (1H and 13C) and bi-dimensional (HSQC) NMR spectroscopy. Finally, large scale chromatographic isolation and depolymerization of each GAG followed by LC/MS and 2D-NMR analysis, allowed the sequences to be defined and components to be evaluated of each GAG including oxidized residues of hexosamines and uronic acids at the reducing ends.
Publisher
MDPI AG,MDPI
Subject
/ Chondroitin Sulfates - analysis
/ Chondroitin Sulfates - chemistry
/ Chromatography, High Pressure Liquid
/ component quantitative analysis
/ Dermatan Sulfate - chemistry
/ Heparitin Sulfate - analysis
/ Heparitin Sulfate - chemistry
/ Intestinal Mucosa - chemistry
/ low molecular weight glycosaminoglycans
/ Magnetic Resonance Spectroscopy
/ orthogonal multi-analytical methods
/ sequence and compositional investigations
/ Swine
