title: Profiling glycol-split heparins by high-performance liquid chromatography/mass spectrometry analysis of their heparinase-generated oligosaccharides.
abstract: Glycol-split (gs) heparins, obtained by periodate oxidation/borohydride reduction of heparin currently used as an anticoagulant and antithrombotic drug, are arousing increasing interest in anticancer and anti-inflammation therapies. These new medical uses are favored by the loss of anticoagulant activity associated with glycol-splitting-induced inactivation of the antithrombin III (AT) binding site. The structure of gs heparins has not been studied yet in detail. In this work, ion pair reversed-phase high-performance liquid chromatography (IPRP-HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) widely used for unmodified heparin has been adapted to the analysis of oligosaccharides generated by digestion with heparinases of gs heparins usually prepared from porcine mucosal heparin. The method was also found to be very effective in analyzing gs derivatives obtained from heparins of different animal and tissue origins. Besides the major 2-O-sulfated disaccharides, heparinase digests of gs heparins contain mainly tetra- and hexasaccharides incorporating one or two gs residues, with distribution patterns typical for individual gs heparins. A heptasulfated, mono-N-acetylated hexasaccharide with two gs residues was shown to be a marker of the gs-modified AT binding site within heparin chains.
authors.affiliation: Ronzoni Institute for Chemical and Biochemical Research, 20133 Milan, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Alekseeva
authors.forename: Benito
authors.initials: B
authors.lastname: Casu
authors.forename: Giangiacomo
authors.initials: G
authors.lastname: Torri
authors.forename: Sabrina
authors.initials: S
authors.lastname: Pierro
authors.forename: Annamaria
authors.initials: A
authors.lastname: Naggi
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D006493:Heparin / Q000737:chemistry* / Q000378:metabolism; D019763:Heparin Lyase / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis*; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013552:Swine
chemical_list: D009844:Oligosaccharides; D010504:Periodic Acid; D006493:Heparin; C009288:metaperiodate; D019763:Heparin Lyase