title: Development of hydrophilic interaction chromatography with quadruple time-of-flight mass spectrometry for heparin and low molecular weight heparin disaccharide analysis.
abstract: BACKGROUND
  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.
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, 215021, China.
authors.forename: Yilan
authors.initials: Y
authors.lastname: Ouyang
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, 215021, China.
authors.forename: Chengling
authors.initials: C
authors.lastname: Wu
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, 215021, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Sun
authors.affiliation: Xiehe Pharmaceutical Co. Ltd, Shijiazhuang, Hebei Province, 050083, China.
authors.forename: Jianfen
authors.initials: J
authors.lastname: Liu
authors.affiliation: Departments of Chemistry and Chemical Biology, Chemical and Biological Engineering, Biomedical Engineering, Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, 215021, China.
authors.forename: Zhenqing
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000925:Anticoagulants / Q000032:analysis / Q000737:chemistry; D002845:Chromatography / Q000379:methods*; D004187:Disaccharides / Q000032:analysis / Q000737:chemistry*; D006493:Heparin / Q000032:analysis* / Q000737:chemistry; D006495:Heparin, Low-Molecular-Weight / Q000032:analysis / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D013058:Mass Spectrometry / Q000379:methods*
chemical_list: D000925:Anticoagulants; D004187:Disaccharides; D006495:Heparin, Low-Molecular-Weight; D006493:Heparin