A screen printed gold electrode (SPGE) served as the foundation for directly depositing Vanadium pentoxide (V2O5), crafting an enzyme-free glucose sensor. Through cyclic voltammetry in an alkaline setting, the sensor's ability to drive glucose oxidation was explored. Utilizing V2O5 as an electrocatalyst, this non-enzymatic sensor exhibited an expansive linear detection range (1 mM–10 mM) and an impressively low detection limit of 0.9 μM. These results underscored V2O5's robust electrocatalytic process in facilitating glucose oxidation within alkaline solutions, unaffected notably by substances like ascorbic acid, fructose and maltose. This investigation highlights a direct and efficient method for glucose detection without reliance on enzymes.