The development of electrochemical sensors with high sensitivity for the simultaneous detection of ascorbic acid (AA), dopamine (DA) and uric acid (UA) is urgently desirable in clinical medicine. However, the challenge lies in achieving simultaneous detection due to their closely oxidation potentials. In this work, we present the synthesis of a composite material comprised of in-situ grown TiO₂ nanowires (NWs) on a Ti₃C₂T_x substrate (Ti₃C₂T_x/TiO₂ NWs) through a facile alkali process. After modifying a glassy carbon electrode (GCE) with Ti₃C₂T_x/TiO₂ NWs (Ti₃C₂T_x/TiO₂ NWs/GCE), it showed excellent electrocatalytic activity for the simultaneous detection of AA/DA/UA by regulating the surface functional groups of Ti₃C₂T_x. Remarkably, the Ti₃C₂T_x/TiO₂ NWs/GCE enabled simultaneous detection of AA in the range of 300-1800 μM, DA in the range of 2-33 μM, and UA in the range of 2-33 μM. The limits of detection (LODs) for AA, DA, and UA were estimated as 66.07 μM, 0.023 μM, and 0.011 μM, respectively. The proposed Ti₃C₂T_x/TiO₂NWs/GCE demonstrated good stability, high selectivity, and reliable reproducibility, making it a promising electrochemical sensor for the detection of AA, DA, and UA. This work offers a new perspective for human health monitoring, paving the way for advancements in this field.