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In this work,a novel kind of heparin-functionalized polyurethane microspheres (Hep-PU MS) was synthesized by a single-step phase separation method.The blood compatibility and anticoagulant effect of the Hep-PU MS were investigated.Then the Hep-PU MS were used to establish an amperometric glucose biosensor that can be applied in whole blood directly,which was based on the antibiofouling property of Hep-PU MS.