Synthesis and characterization of chemically modified hyaluronan and chondroitin sulfate

Mini-Reviews in Organic Chemistry

Volumn 7, Issue 4, 2010, Pages 290-299

  Schiller, J ^a   Becher, J ^b   Moller S ^b   Nimptsch, K ^a   Riemer, T ^a   Schnabelrauch, M ^b  

^a UNIVERSITY HOSPITAL LEIPZIG   (Germany)

^b BIOMATERIALS DEPARTMENT   (Germany)

Author keywords

Chemical functionalization; Chondroitin sulfate; Chromatography; Dermatan sulfate; Glycosaminoglycans; Hyaluronan; Mass spectrometry; NMR spectroscopy

Indexed keywords

CHROMATOGRAPHY; HYALURONIC ACID; NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY; SULFUR COMPOUNDS;

CHEMICAL FUNCTIONALIZATION; CHEMICALLY MODIFIED; CHONDROITINSULFATE; DERMATAN SULPHATE; GLYCOSAMINOGLYCANS; HYALURONAN; NMR-SPECTROSCOPY; PHYSIOLOGICAL FUNCTIONS; SULPHATION; SYNTHESIS AND CHARACTERIZATIONS;

MASS SPECTROMETRY;

EID: 77958181823     PISSN: 1570193X     EISSN: None     Source Type: Journal    
DOI: 10.2174/157019310792246427     Document Type: Article

References (100)

1. Sasisekharan, R.; Raman, R.; Prabhakar, V. Glycomics approach to structure-function relationships of glycosaminoglycans. Annu. Rev. Biomed. Eng., 2006, 8, 181-231.
2. Linhardt, R.J. Analysis of glycosaminoglycans with polysaccharide lyases. Curr. Protoc. Mol. Biol., 2001, Chp. 17: Unit17.13B.
3. Hardingham, T. In: Chemistry and Biology of Hyaluronan; Garg, H.G.; Hales, C.A., Eds.; Elsevier; Amsterdam, 2004; pp. 1-19.
4. Fongmoon, D.; Shetty, A.K.; Basappa, Y.S.; Sugiura, M.; Kongtawelert, P.; Sugahara, K. Chondroitinase-mediated degradation of rare 3-O-sulfated glucuronic acid in functional oversulfated chondroitin sulfate K and E. J. Biol. Chem., 2007, 282, 36895-36904.
5. Sun, D.D.; Guo, X.E.; Likhitpanichkul, M.; Lai, W.M.; Mow, V.C. The influence of the fixed negative charges on mechanical and electrical behaviors of articular cartilage under unconfined compression. J. Biomech. Eng., 2004, 126, 6-16.
6. Fraser, J.R.; Laurent, T.C.; Laurent, U.B. Hyaluronan: its nature, distribution, functions and turnover. J. Intern. Med., 1997, 242, 27-33.
7. Volpi, N.; Schiller, J.; Stern, R.; Soltés, L. Role, metabolism, chemical modifications and applications of hyaluronan. Curr. Med. Chem., 2009, 16, 1718-1745.
8. Kogan G.; Šoltés L.; Stern R.; Schiller J.; Mendichi R. In: Studies in Natural Products Chemistry; Atta-ur-Rahman, Ed.; Elsevier: Amsterdam, 2008; Vol. 34, pp. 789-882.
9. Volpi, N. Chondroitin sulfate: structure, role and pharmacological activity, Academic Press: Amsterdam, 2006.
10. Gama, C.I.; Hsieh-Wilson, L.C. Chemical approaches to deciphering the glycosaminoglycan code. Curr. Opin. Chem. Biol., 2005, 9, 609-619.
11. Lamanna, W.C.; Kalus, I.; Padva, M.; Baldwin, R.J.; Merry, C.L.; Dierks, T. The heparanome - the enigma of encoding and decoding heparan sulfate sulfation. J. Biotechnol., 2007, 129, 290-307.
12. Sugahara, K.; Mikami, T.; Uyama, T.; Mizuguchi, S.; Nomura, K.; Kitagawa, H. Recent advances in the structural biology of chondroitin sulfate and dermatan sulfate. Curr. Opin. Struct. Biol., 2003, 13, 612-620.
13. Linhardt, R.J.; Avci, F.Y.; Toida, T.; Kim, Y.S.; Cygler, M. CS lyases: structure, activity, and applications in analysis and the treatment of diseases. Adv. Pharmacol., 2006, 53, 187-215.
14. Wolfrom, M.L.; Juliano, B.O. Chondroitin sulfate modifications. II. 1 Sulfated and N-deacetylated preparations. J. Am. Chem. Soc., 1960, 82, 2588-2592.
15. Casu, B.; Naggi, A.; Torri, G. Chemical derivatization as a strategy to study structure-activity relationships of glycosaminoglycans. Semin. Thromb. Hemost., 2002, 28, 335-342.
16. Nagasawa, K.; Uchiyama, H.; Wajima, N. Chemical sulfation of preparations of chondroitin 4- and 6-sulfate, and dermatan sulfate. Preparation of chondroitin sulfate alike materials from chondroitin 4-sulfate. Carbohydr. Res., 1986, 158, 183-190.
17. Maruyama, T.; Toida, T.; Imanari, T.; Yu, G.; Linhardt, R.J. Conformational changes and anticoagulant activity of chondroitin sulfate following its Osulfonation. Carbohydr. Res., 1998, 306, 35-43.
18. Barbucci, R.; Leone, G.; Chiumiento, A.; Di Cocco, M.E.; D'Orazio, G.; Gianferri, R.; Delfini, M. Low- and high-resolution nuclear magnetic resonance (NMR) characterisation of hyaluronan-based native and sulfated hydrogels. Carbohydr. Res., 2006, 341, 1848-1858.
19. Magnani, A.; Albanese, A.; Lamponi, S.; Barbucci, R. Blood-interaction performance of differently sulphated hyaluronic acids. Thromb. Res., 1996, 81, 383-395.
20. Hintze, V., Moeller, S., Schnabelrauch, M., Bierbaum, S. Viola, M., Worch, H., Scharnweber, D. Modifications of hyaluronan influence the interaction with human bone morphogenetic protein-4 (hBMP-4). Biomacromolecules, 2009, 10, 3290-3297.
21. Matsuo, M.; Takano, R.; Kamei-Hayashi, K.; Hara, S. A novel regioselective desulfation of polysaccharide sulfates: Specific 6-O-desulfation with N,Obis(trimethylsilyl)acetamide. Carbohydr. Res., 1993, 241, 209-215.
22. Toida, T.; Suzuki, A.; Nakajima, K.; Chaidedgumjorn, A.; Imanari, T. Effect of 6-O-sulfonate hexosamine residue on anticoagulant activity of fully Osulfonated glycosaminoglycans. Glycoconjug. J., 2000, 17, 393-399.
23. McCarthy, M.M.U.; Baker, J.R. Isolation and desulphation of keratan sulphates. Carbohydr. Res., 1979, 69, 151-164.
24. Du, Y.; Taga, A.; Suzuki, S.; Liu, W.; Honda, S. Effect of structure modification of chondroitin sulfate C on its enantioselectivity to basic drugs in capillary electrophoresis. J. Chromatogr. A, 2002, 947, 287-299.
25. Nadkarni, V.D.; Toida, T.; VanGorp, C.L.; Schubert, R.L.; Weiler, J.M.; Hansen, K.P.; Caldwell, E.E.O.; Linhardt, R.J. Preparation and biological activity of N-sulfonated chondroitin and dermatan sulfate derivatives. Carbohydr. Res., 1996, 290, 87-96.
26. Crescenzi, V.; Francescangeli, A.; Renier, D.; Bellini, D. New cross-linked and sulfated derivatives of partially deacetylated hyaluronan: synthesis and preliminary characterization. Biopolymers, 2002, 64, 86-94.
27. Jeanloz, R.W., Forchielli, E. Studies on hyaluronic acid and related substances: I. Preparation of hyaluronic acid and derivatives from human umbilical cord. J. Biol. Chem. 1950, 186, 495-511.
28. Bârzu, T.; Level, M.; Petitou, M.; Lormeau, J.C.; Choay, J.; Schols, D.; Baba, M.; Pauwels, R.; Witvrouw, M.; De Clercq, E. Preparation and anti- HIV activity of O-acylated heparin and dermatan sulfate derivatives with low anticoagulant effect. J. Med. Chem., 1993, 36, 3546-3555.
29. Wada, T.; Chirachanchai, S.; Izawa, N.; Inaki, Y.; Takemoto, K. Synthesis and properties of hyaluronic acid conjugated nucleic acid analogues - 1: Synthesis of deacetylhyaluronan and introduction of nucleic acid bases. J. Bioact. Compat. Polym., 1994, 9, 429-447.
30. Burdick, J.A.; Chung, C.; Jia, X.; Randolph, M.A.; Langer, R. Controlled degradation and mechanical behavior of photopolymerized hyaluronic acid networks. Biomacromolecules, 2005, 6, 386-3891.
31. Möller, S.; Weisser, J.; Bischoff, S.; Schnabelrauch, M. Dextran and hyaluronan methacrylate based hydrogels as matrices for soft tissue reconstruction. Biomol. Eng., 2007, 24, 496-504.
32. Li, Q.; Wang, D.-A.; Elisseeff; H.-J. Heterogeneous-phase reaction of glycidyl methacrylate and chondroitin sulfate: Mechanism of ring-openingtransesterification competition. Macromolecules 2003, 36, 2556-2562.
33. Zhang, M.; James, S.P. Synthesis and properties of melt-processable hyaluronan esters. J. Mater. Sci. Mater. Med., 2005, 16, 587-593.
34. Pouyani, T.; Prestwich, G.D. Functionalized derivatives of hyaluronic acid oligosaccharides: drug carriers and novel biomaterials. Bioconjug. Chem., 1994, 5, 339-347.
35. Charlot, A.; Heyraud, A.; Guenot, P.; Rinaudo, M.; Auzely-Velty, R. Controlled synthesis and inclusion ability of a hyaluronic acid derivative bearing beta-cyclodextrin molecules. Biomacromolecules, 2006, 7, 907-913.
36. Creuzet, C.; Kadi, S.; Rinaudo, M.; Auzély-Velty, R. New associative systems based on alkylated hyaluronic acid. Synthesis and aqueous solution properties. Polymer, 2006, 47, 2706-2713.
37. Shu, X.Z.; Liu, Y.; Luo, Y.; Roberts, M.C.; Prestwich, G.D. Disulfide crosslinked hyaluronan hydrogels. Biomacromolecules, 2002, 3, 1304-1311.
38. Mlcochova, P.; Hajkova, V.; Steiner, B.; Bystricky, S.; Koos, M.; Medova, M.; Velebny, V. Preparation and characterization of biodegradable alkylether derivatives of hyaluronan. Carbohydr. Polym., 2007, 69, 344-352.
39. Serban, M.A.; Yang, G.; Prestwich, G.D. Synthesis, characterization and chondroprotective properties of a hyaluronan thioethyl ether derivative. Biomaterials, 2008, 29, 1388-1399.
40. Möller, S.; Schmidtke, M.; Schnabelrauch, M.; Wutzler, P. Antivirale Kombinationen sowie ihre Verwendung. Patent 2005, DE 10 2005 2004 2643.
41. Jiang, B.; Drouet, E.; Milas, M.; Rinaudo, M. Study on TEMPO-mediated selective oxidation of hyaluronan and the effects of salt on the reaction kinetics. Carbohydr. Res., 2000, 327, 455-461.
42. Weng, L.; Pan, H.; Chen, W. Self-crosslinkable hydrogels composed of partially oxidized hyaluronan and gelatin: in vitro and in vivo responses. J. Biomed. Mater. Res. A, 2008, 85, 352-365.
43. Tomihata, K.; Ikada, Y. Crosslinking of hyaluronic acid with glutaraldehyde. J. Polym. Sci. Pol. Chem., 1997, 35, 3553-3559.
44. Lu, P.L.; Lai, J.Y.; Ma, D.H.K.; Hsiue, G.H. Carbodiimide cross-linked hyaluronic acid hydrogels as cell sheet delivery vehicles: characterization and interaction with corneal endothelial cells. J. Biomater. Sci. Polym. E., 2008, 19, 1-18.
45. 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. A, 1999, 47, 152-169.
46. Kirker, K.R.; Prestwich, G.D. Physical properties of glycosaminoglycan hydrogels. J. Polym. Sci. Pol. Phys., 2004, 42, 4344-4356.
47. Laurent, T.C.; Hellsing, K.; Gelotte, B. Cross-linked gels of hyaluronic acid. Acta Chem. Scand., 1964, 18, 274-275.
48. Segura, T.; Anderson, B.C.; Chung, P.H.; Webber, R.E.; Shull, K.R.; Shea, L.D. Crosslinked hyaluronic acid hydrogels: a strategy to functionalize and pattern. Biomaterials, 2005, 26, 359-371.
49. Wang, S.C.; Chen, B.H.; Wang, L.F.; Chen, J.S. Characterization of chondroitin sulfate and its interpenetrating polymer network hydrogels for sustaineddrug release. Int. J. Pharm., 2007, 329, 103-109.
50. Balazs, E.A.; Leshchiner, A. Cross-linked gels of hyaluronic acid and products containing such gels. Patent 1986, US 4582865.
51. Bodnar, M.; Daroczi, L.; Batta, G.; Bako, J.; Hartmann, J.F.; Borbely, J. Preparation and characterization of cross-linked hyaluronan nanoparticles. Colloid Polym. Sci., 2009, 287, 991-1000.
52. Vercruysse, K.P.; Marecak, D.M.; Marecek, J.F.; Prestwich, G.D. Synthesis and in vitro degradation of new polyvalent hydrazide cross-linked hydrogels of hyaluronic acid. Bioconjug. Chem., 1997, 8, 686-694.
53. Kuo, J.W.; Swann, D.A.; Prestwich, G.D. Chemical modification of hyaluronic acid by carbodiimides. Bioconjug. Chem., 1991, 2, 232-241.
54. Pouyani, T.; Harbison, G.S.; Prestwich, G.D. Novel hydrogels of hyaluronic acid: Synthesis, surface morphology, and solid-state NMR. J. Am. Chem. Soc., 1994, 116, 7515-7522.
55. Shu, X.Z.; Liu, Y.C.; Palumbo, F.S.; Lu, Y.; Prestwich, G.D. In situ crosslinkable hyaluronan hydrogels for tissue engineering. Biomaterials, 2004, 25, 1339-1348.
56. Cai, S.S.; Liu, Y.C.; Shu, X.Z.; Prestwich, G.D. Injectable glycosaminoglycan hydrogels for controlled release of human basic fibroblast growth factor. Biomaterials, 2005, 26, 6054-6067.
57. Huin-Amargier, C.; Marchal, P.; Payan, E.; Netter, P.; Dellacherie, E. New physically and chemically crosslinked hyaluronate (HA)-based hydrogels for cartilage repair. J. Biomed. Mater. Res. A, 2006, 76A, 416-424.
58. Dulong, V.; Lack, S.; Le Cerf, D.; Picton, L.; Vannier, J. P.; Muller, G. Hyaluronan-based hydrogels particles prepared by crosslinking with trisodium trimetaphosphate. Synthesis and characterization. Carbohydr. Polym., 2004, 57, 1-6.
59. Young, J.J.; Cheng, K.M.; Tsou, T.L.; Liu, H.W.; Wang, H.J. Preparation of cross-linked hyaluronic acid film using 2-chloro-1-methylpyridinium iodide or water-soluble 1-ethyl-(3,3-dimethylaminopropyl)carbodiimide. J. Biomater. Sci. Polym. Ed., 2004, 15, 767-780.
60. Ifkovits, J.L.; Burdick, J.A. Review: photopolymerizable and degradable biomaterials for tissue engineering applications. Tissue Eng., 2007, 13, 2369-2385.
61. Miyamoto, K., Sasaki, M., Minamisawa, Y., Kurahashi, Y., Kano, H., Ishikawa, S. Evaluation of in vitro biocompatibility and biodegradation of photocrosslinked hyaluronate hydrogels (HADgels). J. Biomed. Mater. Res. A., 2004, 70A, 550-559
62. Crescenzi, V.; Cornelio, L.; Di Meo, C.; Nardecchia, S.; Lamanna, R. Novel hydrogels via click chemistry: synthesis and potential biomedical applications. Biomacromolecules, 2007, 8, 1844-1850.
63. Shu, X.Z.; Ahmad, S.; Liu, Y.; Prestwich, G.D. Synthesis and evaluation of injectable, in situ crosslinkable synthetic extracellular matrices for tissue engineering. J. Biomed. Mater. Res. A., 2006, 79, 902-912.
64. Nagasawa, K.; Inoue, Y. Reaction between carbohydrates and sulfuric acid. 2. Depolymerization and sulfation of chondroitin sulfate by sulfuric acid. Chem. Pharm. Bull., 1971, 19, 2617-2621.
65. Bienkowski, M.J.; Conrad, H.E. Structural characterization of the oligosaccharides formed by depolymerization of heparin with nitrous acid. J. Biol. Chem., 1985, 260, 356-365.
66. Hassan, M.S.; Mileva, M.M.; Dweck, H.S.; Rosenfeld, L. Nitric oxide products degrade chondroitin sulfates. Nitric Oxide, 1998, 2, 360-365.
67. Tommeraas, K.; Melander, C. Kinetics of hyaluronan hydrolysis in acidic solution at various pH values. Biomacromolecules, 2008, 9, 1535-1540.
68. Toida, T.; Sato, K.; Sakamoto, N.; Sakai, S.; Hosoyama, S.; Linhardt, R.J. Solvolytic depolymerization of chondroitin and dermatan sulfates. Carbohydr. Res., 2009, 344, 888-893.
69. Al-Assaf, S.; Navaratnam, S.; Parsons, B.J.; Phillips, G.O. Chain scission of hyaluronan by peroxynitrite. Arch. Biochem. Biophys., 2003, 411, 73-82.
70. Rees, M.D.; Hawkins, C.L.; Davies, M.J. Hypochlorite-mediated fragmentation of hyaluronan, chondroitin sulfates, and related N-acetyl glycosamines: evidence for chloramide intermediates, free radical transfer reactions, and site-specific fragmentation. J. Am. Chem. Soc., 2003, 125, 13719-13733.
71. Ofman, D.; Slim, G.C.; Watt, D.K.; Yorke, S.C. Free radical induced oxidative depolymerisation of chondroitin sulphate and dermatan sulphate. Carbohydr. Polym., 1997, 33, 47-56.
72. Uchiyama, H.; Dobashi, Y.; Ohkouchi, K.; Nagasawa, K. Chemical change involved in the oxidative reductive depolymerization of hyaluronic acid. J. Biol. Chem., 1990, 265, 7753-7759.
73. Matsumura, G.; Pigman, W. Catalytic role of copper and iron ions in the depolymerization of hyaluronic acid by ascorbic acid. Arch. Biochem. Biophys., 1965, 110, 526-533.
74. Corsaro, M.M.; Pietraforte, D.; Di Lorenzo, A.S.; Minetti, M.; Marino, G. Reaction of peroxynitrite with hyaluronan and related saccharides. Free Radic. Res., 2004, 38, 343-353.
75. Schiller, J.; Fuchs, B.; Arnhold, J.; Arnold, K. Contribution of reactive oxygen species to cartilage degradation in rheumatic diseases: molecular pathways, diagnosis and potential therapeutic strategies. Curr. Med. Chem., 2003, 10, 2123-2145.
76. Mathur, A.M.; Scranton, A.B. Characterization of hydrogels using nuclear magnetic resonance spectroscopy. Biomaterials, 1996, 17, 547-557.
77. Schiller, J.; Fuchs, B.; Arnold, K. The molecular organization of polymers of cartilage: an overview of health and disease. Curr. Org. Chem., 2006, 10, 1771-1789.
78. Schiller, J.; Arnhold, J.; Sonntag, K.; Arnold, K. NMR studies on human, pathologically changed synovial fluids: role of hypochlorous acid. Magn. Reson. Med., 1996, 35, 848-853.
79. Bociek, S.M.; Darke, A.H.; Welti, D.; Rees, D.A. The 13C NMR spectra of hyaluronate and chondroitin sulphates. Further evidence on an alkali-induced conformation change. Eur. J. Biochem., 1980, 109, 447-456.
80. Stylianou, M.; Triantaphyllidou, I.E.; Vynios, D.H. Advances in the analysis of chondroitin/dermatan sulfate. Adv. Pharmacol., 2006, 53, 141-166.
81. Capila, I.; Sasisekharan, R. In: Chemistry and Biology of Hyaluronan; Garg, H.G.; Hales, C.A., Eds.; Elsevier: Amsterdam, 2004; pp. 21-40.
82. Cowman, M.K.; Mendichi, R. In: Chemistry and Biology of Hyaluronan; Garg, H.G.; Hales, C.A., Eds.; Elsevier: Amsterdam: 2004; pp. 41-69.
83. Volpi, N.; Maccari, F.; Linhardt, R.J. Capillary electrophoresis of complex natural polysaccharides. Electrophoresis, 2008, 29, 3095-3106.
84. Mendichi, R.; Soltés, L.; Giacometti Schieroni, A. Evaluation of radius of gyration and intrinsic viscosity molar mass dependence and stiffness of hyaluronan. Biomacromolecules, 2003, 4, 1805-1810.
85. Soltés, L.; Valachová, K.; Mendichi, R.; Kogan, G.; Arnhold, J.; Gemeiner, P. Solution properties of high-molar-mass hyaluronans: the biopolymer degradation by ascorbate. Carbohydr. Res., 2007, 342, 1071-1077.
86. Hascall, V.C.; Midura, R.J.; Sorrell, J.M.; Plaas, A.H. Immunology of chondroitin/dermatan sulfate. Adv. Exp. Med. Biol., 1995, 376, 205-216.
87. Sabiston, P.; Adams, M.E.; Ho, Y.A. Automation of 1,9-dimethylmethylene blue dye-binding assay for sulfated glycosaminoglycans with application to cartilage microcultures. Anal. Biochem., 1985, 149, 543-548.
88. Gold, E.W. The quantitative spectrophotometric estimation of total sulfated glycosaminoglycan levels. Formation of soluble alcian blue complexes. Biochim. Biophys. Acta, 1981, 673, 408-415.
89. Bitter, T.; Muir, H.M. A modified uronic acid carbazole reaction. Anal. Biochem., 1962, 4, 330-334.
90. Frazier, S.B.; Roodhouse, K.A.; Hourcade, D.E.; Zhang, L. The quantification of glycosaminoglycans: A comparison of HPLC, carbazole, and alcian blue methods. Open Glycosci., 2008, 1, 31-39.
91. Prabhakar, V.; Capila, I.; Sasisekharan, R. The structural elucidation of glycosaminoglycans. Methods Mol. Biol., 2009, 534, 147-156.
92. Kreil, G. Hyaluronidases - a group of neglected enzymes. Protein Sci., 1995, 4, 1666-1669.
93. Volpi, N.; Maccari, F. Structural characterization and antithrombin activity of dermatan sulfate purified from marine clam Scapharca inaequivalvis. Glycobiology, 2009, 19, 356-367.
94. Zaia, J.; Li, X.Q.; Chan, S.Y.; Costello, C.E. Tandem mass spectrometric strategies for determination of sulfation positions and uronic acid epimerization in chondroitin sulfate oligosaccharides. J. Am. Soc. Mass Spectrom., 2003, 14, 1270-1281.
95. Busse, K.; Averbeck, M.; Anderegg, U.; Arnold, K.; Simon, J.C.; Schiller, J. The signal-to-noise ratio as a measure of HA oligomer concentration: a MALDI-TOF MS study. Carbohydr. Res., 2006, 341, 1065-1070.
96. Nimptsch, A.; Schibur, S.; Schnabelrauch, M.; Fuchs, B.; Huster, D.; Schiller, J. Characterization of the quantitative relationship between signaltonoise (S/N) ratio and sample amount on-target by MALDI-TOF MS: Determination of chondroitin sulfate subsequent to enzymatic digestion. Anal. Chim. Acta, 2009, 635, 175-182.
97. Ambrosius, M.; Kleesiek, K.; Götting, C. Quantitative determination of the glycosaminoglycan delta-disaccharide composition of serum, platelets and granulocytes by reversed-phase high-performance liquid chromatography. J. Chromatogr. A, 2008, 1201, 54-60.
98. Deakin. J.A.; Yon, M. A simplified and sensitive fluorescent method for disaccharide analysis of both heparan sulfate and chondroitin/dermatan sulfates from biological samples. Glycobiology, 2008, 18, 483-491.
99. Zhang, Z.; Xie, J.; Zhang, F.; Linhardt, R.J. Thin-layer chromatography for the analysis of glycosaminoglycan oligosaccharides. Anal. Biochem., 2007, 371, 118-20.
100. Volpi, N. High-performance liquid chromatography and on-line mass spectrometry detection for the analysis of chondroitin sulfates/hyaluronan disaccharides derivatized with 2-aminoacridone. Anal. Biochem., 2010, 397, 12-23.