Details

Autor(en) / Beteiligte

• Li, Lingyun
• Zhang, Fuming
• Zaia, Joseph
• Linhardt, Robert J

Titel

Top-Down Approach for the Direct Characterization of Low Molecular Weight Heparins Using LC-FT-MS

Ist Teil von

• Analytical chemistry (Washington), 2012-10, Vol.84 (20), p.8822-8829

Ort / Verlag

Washington, DC: American Chemical Society

Erscheinungsjahr

2012

Quelle

MEDLINE

Beschreibungen/Notizen

• Low molecular heparins (LMWHs) are structurally complex, heterogeneous, polydisperse, and highly negatively charged mixtures of polysaccharides. The direct characterization of LMWH is a major challenge for currently available analytical technologies. Electrospray ionization (ESI) liquid chromatography-mass spectrometry (LC-MS) is a powerful tool for the characterization complex biological samples in the fields of proteomics, metabolomics, and glycomics. LC-MS has been applied to the analysis of heparin oligosaccharides, separated by size exclusion, reversed phase ion-pairing chromatography, and chip-based amide hydrophilic interaction chromatography (HILIC). However, there have been limited applications of ESI-LC-MS for the direct characterization of intact LMWHs (top-down analysis) due to their structural complexity, low ionization efficiency, and sulfate loss. Here we present a simple and reliable HILIC-Fourier transform (FT)-ESI-MS platform to characterize and compare two currently marketed LMWH products using the top-down approach requiring no special sample preparation steps. This HILIC system relies on cross-linked diol rather than amide chemistry, affording highly resolved chromatographic separations using a relatively high percentage of acetonitrile in the mobile phase, resulting in stable and high efficiency ionization. Bioinformatics software (GlycReSoft 1.0) was used to automatically assign structures within 5-ppm mass accuracy.