Characterization of epimerization and composition of heparin and dalteparin using a UHPLC-ESI-MS/MS method. (1st January 2019)

• Record Type: Journal Article
• Authors: Zhang, Tianji; Liu, Xin; Li, Hongmei; Wang, Zhangjie; Chi, Lianli; Li, Jin-Ping; Tan, Tianwei
• Is Part Of: Carbohydrate polymers. Volume 203(2018)
• Page Start: 87
• Page End: 94
• Publication Date: 2019-01-01
• Subjects: HONO nitrous acid -- IdoA iduronic acid -- GlcA glucuronic acid -- GlcN glucosamine -- SAX strong anion exchange -- RP-IP reverse phase ion pairing -- HILIC hydrophilic interaction chromatography -- WAX weak anion exchange -- LMWHs low molecular weight heparins -- TDA triple detector array -- ESI electrospray ionization -- MS/MS or MS/MS/MS tandem mass spectrometry -- FT-ICR Fourier transform ion cyclotron resonance -- CID collision induced dissociation -- EDD electron detachment dissociation -- NETD negative electron transfer dissociation -- HCD high energy collision dissociation -- Hsepi D-glucuronyl C5-epimerase -- 6-OST 6-O-sulfotransferase -- HS heparan sulfate -- MEF mouse embryonic fibroblast -- UHPLC ultra high performance LC -- GM GlcA-aManR -- IM IdoA-aManR -- OD O-desulfated -- CFDA China food and drug administration -- PGC porous graphitic carbon; Heparin -- Dalteparin -- Disaccharides -- UHPLC-HILIC/WAX-MS/MS -- Isotopic labelling; Polysaccharides -- Periodicals; Polysaccharides -- Periodicals; Polysaccharides -- Périodiques; Electronic journals; 547.78
• DOI: 10.1016/j.carbpol.2018.08.108
• Languages: English
• ISSNs: 0144-8617

Abstract

Graphical abstract: Highlights: A novel UHPLC-HILIC/WAX-MS/MS approach was applied to characterize HONO depolymerized heparin/dalteparin. Ambiguous disaccharide identification was achieved with unique standards. Isotopic labelling was successfully applied for the sameness evaluation of generic dalteparin. Hexuronic acid epimers were identified by MS/MS/MS (CID and HCD) with LTQ-Orbitrap. Abstract: Heparin is a significant anticoagulant that has been used in clinic over decades. Although numerous efforts have been made to characterize the molecular structure of heparin and its derivatives for safety of the medicine, technical barriers still exist because of their structural complexity. In this study, we have established a method capable to evaluate both the epimerization and composition of heparin and dalteparin by a UHPLC-HILIC/WAX-MS/MS approach. Ten major disaccharide building blocks retaining the epimerization configuration of parental heparin chains were generated and well separated, 9 of which were unambiguously identified. Isomer identifications were achieved through high-resolution tandem mass spectrometry analysis with reference to elaborately prepared standards. The method was successfully applied for the sameness study of generic dalteparins in combination with an isotopic labelling procedure. We believe this robust method maybe adapted to quality control in pharmaceutical production of heparin and LMWHs.