NMR Analysis of Heparin.

Results shown

The standard heparin spectra were obtained using different magnetic fields (400MHz and 500MHz) show that there isn’t any loss of signal nor any significant interference that avoid identification of heparin.

The spectra refer to water suppression techniques. The technique used in the samples’ analysis was pre-saturation, because this was the one that caused the lowest interference in the intensity of the peaks.

The spectrum is an overlap of the standard compounds of Heparin, Sulfated Dermatan, OSCS and Benzoic Alcohol. The region of the proton spectrum between 2.00 p.p.m. and 2.20 p.p.m. determines the identification of these compounds, because N-acetyl group signals is distinct.

It was observed an absence of displacement in the OSCS from these heparin samples, which means that their intake is acceptable. However, it was also observed an absence of DS in some commercial heparin samples.

Literature cited

The following literature references are used for this poster:

  1. Liu, H.; Zhanga, Z.; Linhardt R.J. Lessons learned from the contamination of heparin. Natural Product Reports. 26 (2009) 313–321.
  2. FDA (2008) FDA’s ongoing heparin investigation. http://www.fda.gov/NewsEvents/Testimony/ucm115242.htm
  3. PHARM WHO EXPERT COMMITTEE ON SPECIFICATIONS. http://www.who.int/medicines/publications/pharmprep/pdf_trs953.pdf?ua=1
  4. Guerrini, M.; Beccati, D.; Shriver, Z.; Naggi, A.; Viswanathan. K.; Bisio, A.; Capila, I.; Lansing, J.C.; Guglieri, S.; Fraser, B.; Al-Hakim, A.; Gunay, N. S.; Zhang, Z.; Robinson, L.; Torri, G.; Sasisekharan, R. Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical events. Nature Biotechnology. 26 (2008) 669-675.
  5. Zang, Q.; Keire, D.A.; Wood, R. D.; Buhse, L. F.; Moore, C. M.V.; Nasr, M.; Al-Hakim, A.; Trehyd, M.L.; Welsh, W. J. Combining 1H NMR spectroscopy and chemometrics to identify heparin samples that may possess dermatan sulfate (DS) impurities or oversulfated chondroitin sulfate (OSCS) contaminants. Journal of Pharmaceutical and Biomedical Analysis. 54 (2011) 1020–1029.
  6. Beni, S.; Limtiaco, J. F. K.; Larive, C. K. Analysis and characterization of heparin impurities. Anal Bioanal Chem. 399 (2011) 527–539.
  7. Beyer, T; Diehl, B.; Randel, G.; Humpfer, E.; Schafer, H.; Spraul, M.; Schollmayer, C.; Holzgrabe, U. Quality assessment of unfractionated heparin using 1H nuclear magnetic resonance spectroscopy.
The Poster

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