title: Analysis of 3-O-sulfo group-containing heparin tetrasaccharides in heparin by liquid chromatography-mass spectrometry.
abstract: Complete heparin digestion with heparin lyase 2 affords a mixture of disaccharides and resistant tetrasaccharides with 3-O-sulfo group-containing glucosamine residues at their reducing ends. Quantitative online liquid chromatography-mass spectrometric analysis of these resistant tetrasaccharides is described in this article. The disaccharide and tetrasaccharide compositions of seven porcine intestinal heparins and five low-molecular-weight heparins were analyzed by this method. These resistant tetrasaccharides account for from 5.3 to 7.3wt% of heparin and from 6.2 to 8.3wt% of low-molecular-weight heparin. Because these tetrasaccharides are derived from heparin's antithrombin III-binding sites, we examined whether this method could be applied to estimate the anticoagulant activity of heparin. The content of 3-O-sulfo group-containing tetrasaccharides in a heparin correlated positively (r=0.8294) to heparin's anticoagulant activity.
authors.affiliation: College of Food Science and Technology, Ocean University of China, Qingdao, Shandong 266003, China; Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Guoyun
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Yang
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Lingyun
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Fuming
authors.initials: F
authors.lastname: Zhang
authors.affiliation: College of Food Science and Technology, Ocean University of China, Qingdao, Shandong 266003, China.
authors.forename: Changhu
authors.initials: C
authors.lastname: Xue
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: linhar@rpi.edu.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000494:pharmacology; D000990:Antithrombin III / Q000378:metabolism; D001665:Binding Sites; D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid / Q000379:methods*; D019763:Heparin Lyase / Q000378:metabolism; D006495:Heparin, Low-Molecular-Weight / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D008969:Molecular Sequence Data; D013329:Structure-Activity Relationship; D013552:Swine
chemical_list: D000925:Anticoagulants; D006495:Heparin, Low-Molecular-Weight; D000990:Antithrombin III; D019763:Heparin Lyase
keywords: 3-O-Sulfo group; Anticoagulant activity; Heparin lyase 2; Porcine intestinal heparin; RPIP–LC–MS; Tetrasaccharides