Li Fu1, Lingyun Li2, Chao Cai2, Guoyun Li3, Fuming Zhang4, Robert J Linhardt5. 1. 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. 2. Department of Chemistry and Chemical, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. 3. 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. 4. Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. 5. 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.
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.
The thermal instability of the anticoagulant heparin is associn class="Gene">ated, 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: T Toida; H Yoshida; H Toyoda; I Koshiishi; T Imanari; R E Hileman; J R Fromm; R J Linhardt Journal: Biochem J Date: 1997-03-01 Impact factor: 3.857
Authors: Ujjwal Bhaskar; Anne M Hickey; Guoyun Li; Ruchir V Mundra; Fuming Zhang; Li Fu; Chao Cai; Zhimin Ou; Jonathan S Dordick; Robert J Linhardt Journal: Biotechnol Prog Date: 2015-07-16
Authors: Barbara Mulloy; Nian Wu; Frederick Gyapon-Quast; Lei Lin; Fuming Zhang; Matthew C Pickering; Robert J Linhardt; Ten Feizi; Wengang Chai Journal: Anal Chem Date: 2016-06-15 Impact factor: 6.986