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Article

Characterization of Danaparoid Complex Extractive Drug by an Orthogonal Analytical Approach

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Centro Alta Tecnologia Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni S.r.l., via G. Colombo 81, 20133 Milan, Italy
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Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni S.r.l., via G. Colombo 81, 20133 Milan, Italy
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Aspen Oss B.V., Kloosterstraat 6, 5349 AB Oss, The Netherlands
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Author to whom correspondence should be addressed.
Molecules 2017, 22(7), 1116; https://doi.org/10.3390/molecules22071116
Received: 31 May 2017 / Accepted: 2 July 2017 / Published: 5 July 2017
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. View Full-Text
Keywords: danaparoid sodium; low molecular weight glycosaminoglycans; orthogonal multi-analytical methods; sequence and compositional investigations; component quantitative analysis danaparoid sodium; low molecular weight glycosaminoglycans; orthogonal multi-analytical methods; sequence and compositional investigations; component quantitative analysis
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MDPI and ACS Style

Gardini, C.; Urso, E.; Guerrini, M.; Van Herpen, R.; De Wit, P.; Naggi, A. Characterization of Danaparoid Complex Extractive Drug by an Orthogonal Analytical Approach. Molecules 2017, 22, 1116. https://doi.org/10.3390/molecules22071116

AMA Style

Gardini C, Urso E, Guerrini M, Van Herpen R, De Wit P, Naggi A. Characterization of Danaparoid Complex Extractive Drug by an Orthogonal Analytical Approach. Molecules. 2017; 22(7):1116. https://doi.org/10.3390/molecules22071116

Chicago/Turabian Style

Gardini, Cristina, Elena Urso, Marco Guerrini, René Van Herpen, Pauline De Wit, and Annamaria Naggi. 2017. "Characterization of Danaparoid Complex Extractive Drug by an Orthogonal Analytical Approach" Molecules 22, no. 7: 1116. https://doi.org/10.3390/molecules22071116

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