This study presents a novel sensor for the detection of levodopa (LD) utilizing a glassy carbon electrode modified with a TiO2/MWCNT nanocomposite. Structural analysis confirmed the suitability of the nanocomposite for sensor fabrication, revealing enhancements in effective surface area and sensitivity. Characterization studies employing SEM, EDX, and FT-IR analyses provided insights into the composition and morphology of the modified electrode. The sensor exhibited exceptional performance metrics, including a wide linear detection range (19.6–545 µM), low detection limit (2.51 µM), high repeatability (78.1%), and remarkable average recovery rates in real samples (99.86%). Minimal interference from interfering species fur- ther demonstrated its practical utility. Moreover, the sensor’s direct applicability in diverse sample matrices, without the need for sample separation, highlighted its versatility and convenience. Comparative analysis revealed the sensor’s performance to be comparable to established methods, offering a cost-effective and streamlined approach to LD measurement. Overall, the modified glassy carbon electrode with TiO2/MWCNT nanocomposite presents a clear, suitable, and stable electrocatalytic response, promising significant advancements in biosensing technology for LD detection.