Journal
ANALYTICAL BIOCHEMISTRY
Volume 461, Issue -, Pages 46-48Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2014.05.028
Keywords
Heparin; Sulfate; Stability assay; Amino group; Mass spectrometry
Funding
- NHLBI NIH HHS [RC2 HL101721, R01 HL094463, R01 HL096972, R01 HL062244] Funding Source: Medline
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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-d(6) 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. (C) 2014 Elsevier Inc. All rights reserved.
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