Advertisement

Analytical and Bioanalytical Chemistry

, Volume 410, Issue 18, pp 4259–4273 | Cite as

Separation of hydrophobically modified hyaluronic acid according to the degree of substitution by gradient elution high performance liquid chromatography

  • Carlo Botha
  • Zanelle Viktor
  • Claudine Moire
  • Céline Farcet
  • Fabien Brothier
  • Helen Pfukwa
  • Harald Pasch
Paper in Forefront

Abstract

Amphiphilic hyaluronic acid (HA), synthesised by modifying HA to varying extents with acrylate groups, was successfully separated according to degree of substitution (DS) using solvent gradient high performance liquid chromatography (HPLC). Two HPLC methods based on the amphiphilic structure of the HA were developed. In the first approach, normal phase gradient HPLC was explored, and separation was based on the interactions of HA’s polar hydroxyl groups with a polar cyano stationary phase. In the second approach, separation was based on the interaction of the hydrophobic acrylate moieties with a non-polar C8 stationary phase (reversed phase gradient HPLC). The separation was optimised by using an electrolyte in the sample solvent to suppress non-covalent interactions and improve the selectivity of the developed method. The photolytic stability of the modified and unmodified HA was also investigated in order to optimise the sample preparation procedure. Furthermore, an alternative method to NMR spectroscopy was developed for determining the DS of HA.

Graphical abstract

Keywords

Amphiphilic biopolymers Hyaluronic acid Solvent gradient HPLC Degree of substitution Glycosaminoglycans 

Notes

Acknowledgements

The financial support of this work by L’Oréal Research & Innovation (Aulnay-Sous-Bois, France) is gratefully acknowledged. Bertrand Lion, Julien Portal and Franck Hernandez from L’Oréal laboratories are acknowledged for providing the modified HAs and information related to their synthesis process.

Compliance with ethical standards

The authors declare no conflicts of interest.

Supplementary material

216_2018_1123_MOESM1_ESM.pdf (350 kb)
ESM 1 (PDF 349 kb)

References

  1. 1.
    Xu C, Arancon RAD, Labidid J, Luque R. Lignin depolymerisation strategies: towards valuable chemicals and fuels. Chem Soc Rev. 2014;43:7485–500.CrossRefPubMedGoogle Scholar
  2. 2.
    Azadi P, Inderwildi OR, Farnood R, King DA. Liquid fuels, hydrogen and chemicals from lignin: a critical review. Renew Sust Energ Rev. 2013;21:506–23.CrossRefGoogle Scholar
  3. 3.
    Miller SA. Sustainable polymers: opportunities for the next decade. ACS Macro Lett. 2013;2:550–4.CrossRefGoogle Scholar
  4. 4.
    Delidovich I, Hausoul PJC, Deng L, Pfützenreuter R, Rose M, Palkovits R. Alternative monomers based on lignocellulose and their use for polymer production. Chem Rev. 2016;116:1540–99.CrossRefPubMedGoogle Scholar
  5. 5.
    Roy D, Semsarilar M, Guthrie JT, Perrier S. Cellulose modification by polymer grafting: a review. Chem Soc Rev. 2009;38:2046–64.CrossRefPubMedGoogle Scholar
  6. 6.
    Kuo JW, Swann DA, Prestwich GD. Chemical modification of hyaluronic acid by carbodiimides. Bioconjug Chem. 1991;2:232–41.CrossRefPubMedGoogle Scholar
  7. 7.
    Schanté CE, Zuber G, Herlin C, Vandamme TF. Chemical modifications of hyaluronic acid for the synthesis of derivatives for a broad range of biomedical applications. Carbohyd Polym. 2011;85:469–89.CrossRefGoogle Scholar
  8. 8.
    Singh J, Kaur L, McCarthy OJ. Factors influencing the physico-chemical, morphological, thermal and rheological properties of some chemically modified starches for food applications—a review. Food Hydrocoll. 2007;21:1–22.CrossRefGoogle Scholar
  9. 9.
    Li L, Ni R, Shao Y, Mao S. Carrageenan and its applications in drug delivery. Carbohyd Polym. 2014;103:1–11.CrossRefGoogle Scholar
  10. 10.
    Pelletier S, Hubert P, Lapicque F, Payan E, Dellacherie E. Amphiphilic derivatives of sodium alginate and hyaluronate: synthesis and physico-chemical properties of aqueous dilute solutions. Carbohyd Polym. 2000;43:343–9.CrossRefGoogle Scholar
  11. 11.
    Mayol L, Biondi M, Russo L, Malle BM, Schwach-Abdellaoui K, Borzacchiello A. Amphiphilic hyaluronic acid derivatives toward the design of micelles for the sustained delivery of hydrophobic drugs. Carbohyd Polym. 2014;102:110–6.CrossRefGoogle Scholar
  12. 12.
    Tømmeraas K, Mellergaard M, Malle BM, Skagerlind P. New amphiphilic hyaluronan derivatives based on modification with alkenyl and aryl succinic anhydrides. Carbohyd Polym. 2011;85:173–9.CrossRefGoogle Scholar
  13. 13.
    Pravata L, Braud C, Boustta M, El Ghzaoui A, Tømmeraas K, Guillaumie F, et al. New amphiphilic lactic acid oligomer–hyaluronan conjugates: synthesis and physicochemical characterization. Biomacromolecules. 2008;9:340–8.CrossRefPubMedGoogle Scholar
  14. 14.
    Vasi A-M, Popa MI, Butnaru M, Dodi G, Verestiuc L. Chemical functionalization of hyaluronic acid for drug delivery applications. Mat Sci Eng C. 2014;38:177–85.CrossRefGoogle Scholar
  15. 15.
    Lapčík LJ, Lapčík L, De Smedt S, Demeester J, Chabreček P. Hyaluronan: Preparation, structure, properties, and applications. Chem Rev. 1998;98:2663–84.CrossRefPubMedGoogle Scholar
  16. 16.
    Bulpitt P, Aeschlimann D. New strategy for chemical modification of hyaluronic acid: preparation of functionalized derivatives and their use in the formation of novel biocompatible hydrogels. J Biomed Mater Res. 1999;47:152–69.CrossRefPubMedGoogle Scholar
  17. 17.
    Eenschooten C, Guillaumie F, Kontogeorgis GM, Stenby EH, Schwach-Abdellaoui K. Preparation and structural characterisation of novel and versatile amphiphilic octenyl succinic anhydride–modified hyaluronic acid derivatives. Carbohyd Polym. 2010;79:597–605.CrossRefGoogle Scholar
  18. 18.
    Prestwich GD, Marecak DM, Marecek JF, Vercruysse KP, Ziebell MR. Controlled chemical modification of hyaluronic acid: synthesis, applications, and biodegradation of hydrazide derivatives. J Control Release. 1998;53:93–103.CrossRefPubMedGoogle Scholar
  19. 19.
    Christner JE, Brown ML, Dziewiatkowski DD. Interaction of cartilage proteoglycans with hyaluronic acid. The role of the hyaluronic acid carboxyl groups. Biochem J. 1977;167:711–6.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Jia X, Colombo G, Padera R, Langer R, Kohane DS. Prolongation of sciatic nerve blockade by in situ cross-linked hyaluronic acid. Biomaterials. 2004;25:4797–804.CrossRefPubMedGoogle Scholar
  21. 21.
    Oh EJ, Kang S-W, Kim B-S, Jiang G, Cho IH, Hahn SK. Control of the molecular degradation of hyaluronic acid hydrogels for tissue augmentation. J Biomed Mater Res A. 2008;86A:685–93.CrossRefGoogle Scholar
  22. 22.
    Ghosh P, Guidolin D. Potential mechanism of action of intra-articular hyaluronan therapy in osteoarthritis: are the effects molecular weight dependent? Semin Arthritis Rheum. 2002;32:10–37.CrossRefPubMedGoogle Scholar
  23. 23.
    Stern R, Asari AA, Sugahara KN. Hyaluronan fragments: an information-rich system. Eur J Cell Biol. 2006;85:699–715.CrossRefPubMedGoogle Scholar
  24. 24.
    Oudshoorn MHM, Rissmann R, Bouwstra JA, Hennink WE. Synthesis of methacrylated hyaluronic acid with tailored degree of substitution. Polymer. 2007;48:1915–20.CrossRefGoogle Scholar
  25. 25.
    Wende FJ, Gohil S, Mojarradi H, Gerfaud T, Nord LI, Karlsson A, et al. Determination of substitution positions in hyaluronic acid hydrogels using NMR and MS based methods. Carbohyd Polym. 2016;136:1348–57.CrossRefGoogle Scholar
  26. 26.
    Falkenhagen J, Much H, Stauf W, Müller AHE. Characterization of block copolymers by liquid adsorption chromatography at critical conditions. 1. Diblock copolymers. Macromolecules. 2000;33:3687–93.CrossRefGoogle Scholar
  27. 27.
    Gorshkov AV, Much H, Becker H, Pasch H, Evreinov VV, Entelis SG. Chromatographic investigations of macromolecules in the critical range of liquid chromatography. J Chromatogr A. 1990;523:91–102.CrossRefGoogle Scholar
  28. 28.
    Glöckner G. Quantitative aspects of gradient HPLC of copolymers from styrene and ethyl methacrylate. Chromatographia. 1987;23:517–24.CrossRefGoogle Scholar
  29. 29.
    Barth HG, Boyes BE, Jackson C. Size exclusion chromatography. Anal Chem. 1996;68:445–66.CrossRefGoogle Scholar
  30. 30.
    Trathnigg B. Size-exclusion chromatography of polymers. In: Meyers RA, editor. Encyclopedia of analytical chemistry. Chichester: John Wiley & Sons, Ltd; 2000. pp. 8008–34.Google Scholar
  31. 31.
    Stenekes RJH, Hennink WE. Polymerization kinetics of dextran-bound methacrylate in an aqueous two phase system. Polymer. 2000;41:5563–9.CrossRefGoogle Scholar
  32. 32.
    Smeds KA, Pfister-Serres A, Miki D, Dastgheib K, Inoue M, Hatchell DL, et al. Photocrosslinkable polysaccharides for in situ hydrogel formation. J Biomed Mater Res A. 2001;54:115–21.CrossRefGoogle Scholar
  33. 33.
    Guiochon G, Moysan A, Holley C. Influence of various parameters on the response factors of the evaporative light scattering detector for a number of non-volatile compounds. J Liq Chromatogr. 1988;11:2547–70.CrossRefGoogle Scholar
  34. 34.
    Mathews BT, Higginson PD, Lyons R, Mitchell JC, Sach NW, Snowden MJ, et al. Improving quantitative measurements for the evaporative light scattering detector. Chromatographia. 2004;60:625–33.CrossRefGoogle Scholar
  35. 35.
    Arndt JH, Macko T, Brüll R. Application of the evaporative light scattering detector to analytical problems in polymer science. J Chromatogr A. 2013;1310:1–14.CrossRefPubMedGoogle Scholar
  36. 36.
    Bashir MA, Brüll A, Radke W. Fast determination of critical eluent composition for polymers by gradient chromatography. Polymer. 2005;46:3223–9.CrossRefGoogle Scholar
  37. 37.
    Brun P, de Galateo A, Camporese A, Cortivo R, Abatangelo G. Analysis of hyaluronic acid in synovial fluid by reversed-phase liquid chromatography. J Chromatogr B Biomed Sci Appl. 1990;526:530–4.CrossRefGoogle Scholar
  38. 38.
    Jiang X, Horst A, Schoenmakers PJ. Breakthrough of polymers in interactive liquid chromatography. J Chromatogr A. 2002;982:55–68.CrossRefPubMedGoogle Scholar
  39. 39.
    Reingruber E, Bedani F, Buchberger W, Schoenmakers P. Alternative sample-introduction technique to avoid breakthrough in gradient-elution liquid chromatography of polymers. J Chromatogr A. 2010;1217:6595–8.CrossRefPubMedGoogle Scholar
  40. 40.
    Reháková M, Bakoš D, Soldán M, Vizárová K. Depolymerization reactions of hyaluronic acid in solution. Int J Biol Macromol. 1994;16:121–4.CrossRefPubMedGoogle Scholar
  41. 41.
    Adrian J, Esser E, Hellmann G, Pasch H. Two-dimensional chromatography of complex polymers part 1. Analysis of a graft copolymer by two-dimensional chromatography with on-line FTIR detection. Polymer. 2000;41:2439–49.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Carlo Botha
    • 1
  • Zanelle Viktor
    • 1
  • Claudine Moire
    • 2
  • Céline Farcet
    • 2
  • Fabien Brothier
    • 2
  • Helen Pfukwa
    • 1
  • Harald Pasch
    • 1
  1. 1.Department of Chemistry and Polymer ScienceStellenbosch UniversityMatielandSouth Africa
  2. 2.L’Oréal Research and InnovationAulnay-Sous-BoisFrance

Personalised recommendations