Four hyaluronan based hydrogels with different cross-linking agents were synthesised. Compounds with an increasing hydrophobic character and length as spacer arms were chosen, i.e. O,O'-bis(2 aminopropyl)polyethylen glicol, 1.3 diaminopropane, 1.6 diaminohexane, and 1.12 diaminedodecane. The cross-linking reaction involved 50% of the carboxylate groups present in the hyaluronan macromolecule chain. Hydrogels were characterised by water-uptake measurements and SEM analysis. The adsorption of two plasma proteins (HSA and Fbg) was analysed onto the four hydrogels in flow conditions by ATR-FTIR, evaluating the adsorption kinetics and the eventual protein conformational changes. The increasing hydrophobic character of the surface imposes a clear trend to the protein adsorption kinetics. The influence of the spacer arms on the protein conformation is evident on HSA, whereas Fbg does not seem to be significantly influenced by the change of the substrate hydrophobic properties.