The reported work discussed the simpler and sensitive strategy for the electrochemical determination of furosemide by employed tranexamic acid derived gold nanoparticles modified glassy carbon electrode (GCE). The synthesis of tranexamic acid derived gold nanoparticles (Tr-AuNps) was carried out using single step approach. The synthesized Tr-AuNps were characterized by using atomic force microscopy (AFM), illustrated that the particles are spherical in shape with an average size of 35 nm. The synthesized AuNps have modified the sensing surface of GCE. The modified GCE demonstrated highly catalytic behavior for the oxidation of loop diuretic drug furosemide. The influence of pH and supporting electrolyte was examined and the working conditions were optimized. The amperometric determination of furosemide was also carried out at the Tr-AuNps modified GCE under stirred conditions using Britton Robinson buffer (BR buffer) as supporting electrolyte at pH(5). The linear calibration plot showed the dependence of the peak current on increasing concentrations of furosemide in the range of 50 mu M to 500 mu M furosemide with the detection limit of 5 mu M. The proposed sensing plan has been successfully employed for the quantification of furosemide in human urine samples with satisfactory recoveries.