Preparation and characterisation of copper(II) hyaluronate

Amorphous copper complexes of the general composition Cu(C14H20O11N)2 x xH2O have been prepared with high- and low-molecular-weight hyaluronic acid (HA). Optimal conditions for preparation are obtained at pH values from 5.0 to 5.5, with a molar ratio of HA versus Cu2+ of 1:1, and at a mass concentration of 5 and 10 mg/mL for high- (Mw = 1.8 x 10(6) Da) and low-molecular-weight sodium hyaluronate (Mw = 2 x 10(5) Da), respectively. The coordination polyhedron of the copper ion has been elucidated by EXAFS and XANES spectroscopy. Copper atoms are octahedrally coordinated in both cases with four equatorial Cu-O bond lengths of 1.95 A, and two axial Cu-O bonds of 2.46 A. Visible spectra of acidic aqueous solution suggest that substitution of axial oxygens by NH groups occurs at pH 6.5 or higher. If the pH value of the copper(II) hyaluronate solution increases above 6.5, the coordination of copper(II) changes. It is very likely that the N atom coming from the acetamido group enters into the coordination sphere of the copper(II) ion.