Hydrogel materials have good biomimetic properties and high potential for biomedical and bioanalytical applications. In this work, an hydrogel of serum albumin crosslinked with poly-(ethylene glycol) was prepared and characterized for its water content, protein structure and stability. The ability of the hydrogel to bind small molecule ligands with different hydrophobicity was evaluated using a homologous series of amphiphiles (NBD-Cn, n=4, 6 and 8) and the calculated binding affinities were similar to that of free protein in solution. Overall, the results indicate this type of hydrogel system as convenient tools for studying the binding of xenobiotics to tissue proteins.