Heparin Isomeric Oligosaccharide Separation Using Volatile Salt Strong Anion Exchange Chromatography



Miller, RL, Guimond, SE, Shivkumar, M, Blocksidge, J, Austin, JA, Leary, JA and Turnbull, JE ORCID: 0000-0002-1791-754X
(2016) Heparin Isomeric Oligosaccharide Separation Using Volatile Salt Strong Anion Exchange Chromatography. Analytical Chemistry, 88 (23). 11542 - 11550.

This is the latest version of this item.

[img] Text
Miller et al Analyt Chem 2016 VS-CTA SAX .pdf - Accepted Version

Download (866kB)
[img] Text
Miller et al 2016 Analyt Chem VSCTA-SAX Author accepted.pdf - Accepted Version

Download (999kB)

Abstract

The complexity of heparin and heparan sulfate saccharides makes their purification, including many isomeric structures, very challenging and is a bottleneck for structure–activity studies. High-resolution separations have been achieved by strong anion exchange (SAX) chromatography on Propac PA1 and cetyltrimethylammonium (CTA)-C18 silica columns; however, these entail subsequent desalting methodologies and consequent sample losses and are incompatible with orthogonal chromatography methodologies and, in particular, mass spectrometry. Here, we present the CTA-SAX purification of heparin oligosaccharides using volatile salt (VS) buffer. In VSCTA-SAX, the use of ammonium bicarbonate buffer for elution improves resolution through both weaker dissociation and conformational coordination of the ammonium across the sulfate groups. Using ion mobility mass spectrometry, we demonstrate that isomeric structures have different structural conformations, which makes chromatographic separation achievable. Resolution of such structures is improved compared to standard SAX methods, and in addition, VSCTA-SAX provides an orthogonal method to isolate saccharides with higher purity. Because ammonium bicarbonate is used, the samples can be evaporated rather than desalted, preventing substantial sample loss and allowing more effective subsequent analysis by electrospray mass spectrometry. We conclude that VSCTA-SAX is a powerful new tool that helps address the difficult challenge of heparin/heparan sulfate saccharide separation and will enhance structure–activity studies.

Item Type: Article
Uncontrolled Keywords: Chemical structure, Cations, Heparin, Oligosaccharides Ions
Depositing User: Symplectic Admin
Date Deposited: 20 Mar 2017 10:07
Last Modified: 26 Nov 2020 08:11
DOI: 10.1021/acs.analchem.6b02801
Related URLs:
URI: http://livrepository.liverpool.ac.uk/id/eprint/3006520

Available Versions of this Item