In this paper, we report the use of glassy carbon electrode for electrochemical oxidation of glucose in basic medium. The effect of glucose concentration, scan rate, and point potential was studied. Heterogeneous electron transfer parameters such as standard electron transfer rate constant (k0), transfer coefficient (α), and E0 were calculated using cyclic voltammetry (CV), semi integration of cyclic voltammetry (SICV), semi differentiation of cyclic voltammetry (SDCV), and digital simulation techniques. Amperometric experiments were performed for determination of glucose concentration in basic media. These experiments show a broad linear concentration range, up to 11 mM with a sensitivity of 927.96 μAmM−1cm−2. The lower detection limit (LOD) and Lower Quantification limit (LOQ) were found to be 50 μM and 0.5 mM, respectively. Effect of interfering species such as uric acid, ascorbic acid, acetaminophen, and chloride ion on the electrocatalytic behavior of glassy carbon electrode (GCE) was also studied.