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Structural analysis of heparin by raman spectroscopy
D H Atha1, A K Gaigalas, V Reipa
1Biotechnology Division, National Institute of Standards and Technology, Gaithesburg, MD 20899, USA.
Journal of Pharmaceutical Sciences
|January 1, 1996
Summary
Raman spectroscopy identifies specific sulfate groups in heparin, crucial for its biological activity and quality control of pharmaceutical preparations. This technique aids in determining the proportion of N-sulfate, 6-O-sulfate, and 2-O-sulfate groups.
Area of Science:
- Biochemistry
- Spectroscopy
- Pharmaceutical Science
Background:
- Heparin's biological activity is linked to its specific sulfation patterns.
- Accurate characterization of heparin's structure is vital for quality assurance.
Purpose of the Study:
- To utilize Raman spectroscopy for identifying and quantifying specific sulfate groups in heparin.
- To explore the utility of Raman spectroscopy in heparin quality control.
Main Methods:
- Analysis of Raman spectra of heparin disaccharide standards and porcine mucosal heparin.
- Assignment of characteristic Raman bands to N-sulfate, 6-O-sulfate, and 2-O-sulfate groups.
- Support of spectral assignments using semiempirical quantum mechanical calculations.
Main Results:
- Identified distinct Raman bands for N-sulfate (1039 cm-1), 6-O-sulfate (1055 cm-1), and 2-O-sulfate (1065 cm-1) groups.
- Determined the relative proportions of these sulfate groups in commercial heparin.
- Demonstrated the complementary nature of Raman spectroscopy to NMR for heparin structural analysis.
Conclusions:
- Raman spectroscopy is effective for characterizing heparin's sulfation patterns.
- This method provides crucial structural information for assessing heparin's biological activity.
- Raman spectroscopy enhances the quality assurance of pharmaceutical heparin preparations.