Abstract Glutathione-chitosan conjugates are adequate carriers for anticancer treatment due to their ability for inhibition of efflux pumps, improved mucoadhesivity and in-situ gelling. These conjugates were obtained via carbodiimide at different reaction times in order to get different thiolation degrees ( i.e. , 4.4!, 5.1! and 7.0!) and their behavior in aqueous media at a pH of 4, 5 and 6 was assessed by measurements of hydrodynamic diameter, zeta potential and rheological analyses at a pH ranging from 4 to 6. Data examination was conducted by principal component analysis (PCA) which in turn explained 73.1! of data variability. All samples showed a Newtonian flow and thiolation rendered materials with a highly pronounced temperature-dependent behavior and a gel-like structure. In turn, the phase shift angle was the most prominent rheological change especially at a pH of 5.0 and 6.0 due to the formation of disulfide bonds. The thiolation degree was the most influential factor and it was inversely related to particle charge and consistency index.