The interactions between sodium hyaluronate, an anionic polysaccharide, with surfactants (anionic and nonionic) were investigated using pyrene fluorescence measurement methods. The change of micropolarity produced by the interaction was monitored by the measurement of emission intensity ratio between the first and third bands (I 1 /I 3 ), and the intensity ratio of the excimer and the third vibration monomer band (I E /I M ). Because the hydrophilic heads on the SDS were attracted by the domains formed by the hydroxyl groups of hyaluronate, the I 1 /I 3 ratio was reduced by the addition of hyaluronate at lower than 0.06% of sodium dodecyl sulfate (SDS) concentration. No aggregation was observed between hyaluronate and nonionic surfactants (Tween-80 and Cremophor EL) in the whole concentration range studied. At a higher concentration of surfactant, the I 1 /I 3 ratio of hyaluronate/surfactant was influenced by the addition of saccharide (glucose, lactose, or mannitol). However, the effect of saccharide could be reduced by the addition of salt.