Improved impurity fingerprinting of heparin by high resolution (1)H NMR spectroscopy.
For improving the identification of potential heparin impurities such as oversulfated chondroitin sulfate (OSCS) the standard 2D (1)H-(1)H NMR NOESY was applied. Taking advantage of spin diffusion and adjusting the experimental parameters accordingly additional contaminant-specific signals of the corresponding sugar ring protons can easily be detected. These are usually hidden by the more intense heparin signals. Compared to the current 1D (1)H procedure proposed for screening commercial unfractionated heparin samples and focusing on the contaminants acetyl signals more informative and unique fingerprints may be obtained. Correspondingly measured (1)H fingerprints of a few potential impurities are given and their identification in two contaminated commercial heparin samples is demonstrated. The proposed 2D NOESY method is not intended to replace the current 1D method for detecting and quantifying heparin impurities but may be regarded as a valuable supplement for an improved and more reliable identification of these contaminants.
Bigler
Peter
P
University of Berne, Department of Chemistry and Biochemistry, Laboratory for NMR Spectroscopy, Bern, Switzerland. peter.bigler@ioc.unibe.ch
Brenneisen
Rudolf
R
D000215:Acylation; D000925:Anticoagulants / Q000032:analysis*; D003871:Dermatan Sulfate / Q000032:analysis; D004340:Drug Contamination; D006493:Heparin / Q000032:analysis*; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D013431:Sulfates / Q000032:analysis
D000925:Anticoagulants; D007202:Indicators and Reagents; D013431:Sulfates; D003871:Dermatan Sulfate; D006493:Heparin