Hyaluronidase　(HAase)　is　a　potential　tumor　biomarker　for　diseases　of　the　digestive　tract　and　nervous　system,　the　development　of　simple　and　sensitive　techniques　for　HAase　determination　is　urgent　needed.　Gold　nanorods　(Au　NRs)　can　be　etched　by　H2O2　with　high　efficiency　and　display　color　changing.　In　this　work,　a　HAase-responsive　hydrogel　system　had　been　designed　and　the　amount　of　H2O2　spilled　from　the　system　had　a　close　relationship　with　the　amount　of　HAase,　then　the　spilled　H2O2　had　been　applied　to　etch　Au　NRs.　The　color　change　of　the　solution　was　used　to　realize　semi-quantitative　determination　of　HAase.　Furthermore,　the　longitudinal　peak　shift　of　Au　NRs　had　a　linear　correlation　with　the　concentration　of　HAase　in　the　range　of　10-60　U/mL　(within　40　min)　and　the　limit　of　detection　(LOD)　was　3.8　U/mL　(S/N　=　3),　which　can　be　used　to　realize　accurate　quantitative　analysis　of　HAase.　The　proposed　method　has　been　applied　to　monitor　HAase　in　serum　of　pancreatic　cancer　patients　with　satisfied　results.