Advertisement

Heparin composition: calculation based on elemental analysis and NMR data

  • Ivan ŠimkovicEmail author
  • Michal Raab
  • Raniero Mendichi
  • Alena Manová
  • Alberto Giacometti Schieroni
  • Miloš Hricovíni
Original Paper
  • 27 Downloads

Abstract

The analysis of many polysaccharides is often complicated due to the heterogeneity in their structures. A computer approach is presented exploiting the experimental data from elemental analysis and high-resolution NMR experiments to estimate the individual anhydro-unit ratio. Our approach is based on the fact that elemental composition of a particular type of saccharide is the same for saccharide units, connected by glycosidic bonds forming the polysaccharide chain, or for the individual saccharide anhydro structures complemented by the relevant amount of water molecules. From the possible structures generated, the structure is chosen by the computation with the best fit (“score”) to the elemental composition. The method was tested on a heparin pentasaccharide using the 1% deviation of HSQC-integrated limits at 5% water content. The optimal limits for evaluation of heparin in various ionic forms (Na+, Li+, or NH4+) for the computation input were found by the HSQC integration of the anomeric signals and anhydro unit ratios calculated based on elemental analysis, AAS, and SEC-MALS analysis. The results of heparin from different sources, and in various ionic environments, indicate that the presented methodology could be used as the complementary approach to other experimental analytical techniques of heparin and heparin sulfate.

Graphic abstract

Keywords

Heparin Computer program Elemental analysis NMR AAS SEC-MALS 

Notes

Acknowledgements

We gratefully acknowledge the support provided by the Slovak Granting Agency VEGA (Project # 2/0022/18 and 1/0489/16).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11696_2019_957_MOESM1_ESM.docx (142 kb)
Supplementary material 1 (DOCX 141 kb)

References

  1. Heikkinen S, Toikka MM, Karhunen PT, Kilpeläinen IA (2003) Quantitative 2D HSQC (Q-HSQC) via suppression of J-dependence of polarization transfer in NMR spectroscopy: application to wood lignin. J Amer Chem Soc 125:4363–4367CrossRefGoogle Scholar
  2. Horton D, Pardoe WD (1970) Calculation of degree of substitution of polysaccharides: a FORTRAN computer program. Carbohydr Res 12:269–272CrossRefGoogle Scholar
  3. Hricovíni M (2015) Solution structure of heparin pentasaccharide: NMR and DFT analysis. J Phys Chem 119:12397–12409.  https://doi.org/10.1021/acs.jpcb.5b07046 CrossRefGoogle Scholar
  4. Hricovíni M, Torri G (1995) Dynamics in aqueous solution of the pentasaccharide corresponding to the binding site of heparin for antithrombin III studied by NMR relaxation measurements. Carbohydr Res 268:159–175CrossRefGoogle Scholar
  5. Manová A, Beinrohr E, Čacho F (2017) Atomic absorption spectrometry with electrothermal atomization to determine trace amounts of arsenic. Acta Chim Slov 10:175–179.  https://doi.org/10.1515/acs-2017-0029 CrossRefGoogle Scholar
  6. Mauri L, Boccacio G, Torri G, Karfunkle M, Macchi E, Muzi L, Keire D, Guerrini M (2017) Qualification of HSQC methods for quantitative composition of heparin and low molecular heparins. J Pharm Biomed Anal 136:92–105.  https://doi.org/10.1016/j.lpba.2016.12.031 CrossRefPubMedGoogle Scholar
  7. Mourier P, Anger P, Martinez C, Herman F, Viskov C (2015) Quantitative compositional analysis of heparin using exhaustive heparinase digestion and strong anion exchange chromatography. Anal Chem Res 3:46–53.  https://doi.org/10.1016/j.ancr.2014.12.001 CrossRefGoogle Scholar
  8. Nguyen K, Rabenstein DL (2011) Determination of the primary structure and carboxyl PkAs of heparin-derived oligosaccharides by band-selective homonuclear-decoupled two-dimensional 1H NMR. Anal Bioanal Chem 399:663–671.  https://doi.org/10.1007/s00216-010-4224-4 CrossRefPubMedGoogle Scholar
  9. Petitou M, Duchaussoy P, Lederman I, Choay J, Jacquinet JC, Sinay P, Torri G (1987) Synthesis of heparin fragments: a methyl α-pentaoside with high affinity for antithrombin III. Carbohydr Res 167:67–75CrossRefGoogle Scholar
  10. Šimkovic I (2013) Unexplored possibilities of all-polysaccharide composites. Carbohydr Polym 95:697–715.  https://doi.org/10.1016/j.carbpol.2013.03.040 CrossRefPubMedGoogle Scholar
  11. Šimkovic I, Mendichi R, Kelnar I, Filip J, Hricovíni M (2015) Cationization of heparin for film applications. Carbohyd Polym 115:551–558.  https://doi.org/10.1016/j.carbpol.2014.09.021 CrossRefGoogle Scholar
  12. Šimkovic I, Kelnar I, Mendichi R, Bertok T, Filip J (2017) Composite films prepared from agricultural by-products. Carbohydr Polym 156:77–85.  https://doi.org/10.1016/j.carbpol.2016.09.014 CrossRefPubMedGoogle Scholar
  13. Šimkovic I, Kelnar I, Mendichi R, Tracz A, Filip J, Bertok T, Kasak P (2018) Interfaces study of all-polysaccharide composite films. Chem Pap 72:711–718CrossRefGoogle Scholar
  14. Wang B, Buhse LF, Al-Hakim A, Boyne Ii MT, Keire DA (2012) Characterization of currently marked heparin products: analysis of heparin digests by RPIP-UHPLC-QTOF-MS. J Pharm Biomed Anal 67–68:42–50.  https://doi.org/10.1016/j.jpba.2012.04.033 CrossRefPubMedGoogle Scholar
  15. Yang X, Du H, Liu J, Zhai G (2015) Advanced nanocarriers based on heparin and its derivatives for cancer management. Biomacromol 16:423–434.  https://doi.org/10.1021/bm501532e CrossRefGoogle Scholar

Copyright information

© Institute of Chemistry, Slovak Academy of Sciences 2019

Authors and Affiliations

  1. 1.Institute of ChemistrySlovak Academy of SciencesBratislavaSlovakia
  2. 2.HighChem Ltd.BratislavaSlovakia
  3. 3.Istituto per lo Studio delle Macromolecole (CNR)MilanItaly
  4. 4.Institute of Analytical ChemistrySlovak Technical UniversityBratislavaSlovakia

Personalised recommendations