Development of hydrophilic interaction chromatography with quadruple-time-of-flight mass spectrometry for heparin and low molecular weight heparin disaccharide analysis

Authors
Ouyang, Yilan
Wu, Chengling
Sun, Xue
Liu, Jianfen
Linhardt, Robert J.
Zhang, Zhenqing
ORCID
https://orcid.org/0000-0003-2219-5833
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Other Contributors
Issue Date
2016-01-30
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Full Citation
Development of hydrophilic interaction chromatography with quadruple-time-of-flight mass spectrometry for heparin and low molecular weight heparin disaccharide analysis, C. Wu, Y. Ouyang, X. Sun, N. Xu, R. J. Linhardt, Z. Zhang, Rapid Communications in Mass Spectrometry, 30, 277-284, 2015.
Abstract
Rationale: Heparin and low molecular weight heparin (LMWH) are widely used as clinical anticoagulants. The determination of their composition and structural heterogeneity still challenges analysts. Methods: Disaccharide compositional analysis, utilizing heparinase-catalyzed depolymerization, is one of the most important ways to evaluate the sequence, structural composition and quality of heparin and LMWH. Hydrophilic interaction chromatography coupled with quadruple time-of-flight mass spectrometry (HILIC/QTOFMS) has been developed to analyze the resulting digestion products. Results: HILIC shows good resolution and excellent MS compatibility. Digestion products of heparin and LMWHs afforded up to 16 compounds that were separated using HILIC and analyzed semi-quantitatively. These included eight common disaccharides, two disaccharides derived from chain termini, three 3-O-sulfo-group-containing tetrasaccharides, along with three linkage region tetrasaccharides and their derivatives. Structures of these digestion products were confirmed by mass spectral analysis. The disaccharide compositions of a heparin, two batches of the LMWH, enoxaparin, and two batches of the LMWH, nadroparin, were compared. In addition to identifying disaccharides, 3-O-sulfo-group-containing tetrasaccharides, linkage region tetrasaccharides were observed having slightly different compositions and contents in these heparin products suggesting that they had been prepared using different starting materials or production processes. Conclusions: Thus, compositional analysis using HILIC/QTOFMS offers a unique insight into different heparin products.
Description
Rapid Communications in Mass Spectrometry, 30, 277-284
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Department
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
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Relationships
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
Rapid Communications in Mass Spectrometry
https://harc.rpi.edu/
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https://login.libproxy.rpi.edu/login?url=https://doi.org/10.1002/rcm.7437