title: Heparin stability by determining unsubstituted amino groups using hydrophilic interaction chromatography mass spectrometry.
abstract: The thermal instability of the anticoagulant heparin is associated, in part, with the solvolytic loss of N-sulfo groups. This study describes a new method to assess the increased content of unsubstituted amino groups present in thermally stressed and autoclave-sterilized heparin formulations. N-Acetylation of heparin samples with acetic anhydride-d6 is followed by exhaustive heparinase treatment and disaccharide analysis by hydrophilic interaction chromatography mass spectrometry (HILIC-MS). The introduction of a stable isotopic label provides a sensitive probe for the detection and localization of the lost N-sulfo groups, potentially providing valuable insights into the degradation mechanism and the reasons for anticoagulant potency loss.
authors.affiliation: Department of Chemistry and Chemical, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Fu
authors.affiliation: Department of Chemistry and Chemical, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Lingyun
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Chemistry and Chemical, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Chao
authors.initials: C
authors.lastname: Cai
authors.affiliation: Department of Chemistry and Chemical, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; College of Food Science and Technology, Ocean University of China, Qingdao, Shandong 266003, China.
authors.forename: Guoyun
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Fuming
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Chemistry and Chemical, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Biomedical Engineering, 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: D000588:Amines / Q000737:chemistry*; D000925:Anticoagulants / Q000737:chemistry*; D002626:Chemistry, Pharmaceutical; D002845:Chromatography / Q000379:methods*; D004187:Disaccharides / Q000032:analysis; D004355:Drug Stability; D006493:Heparin / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D013058:Mass Spectrometry / Q000379:methods*
chemical_list: D000588:Amines; D000925:Anticoagulants; D004187:Disaccharides; D006493:Heparin
keywords: Amino group; Heparin; Mass spectrometry; Stability assay; Sulfate