In this research work, a straightforward strategy for the development of carbon-based electrodes is presented using nanoclay as a modifier, and a room temperature ionic liquid was employed as a binder for the simultaneous determination of dopamine (DP) and uric acid (UA) in the presence of ascorbic acid (AA) in real samples in the form of serum and urine. The synergistic effect between nanoclay and the ionic liquid results in an increase in the activity of the prepared carbon ionic liquid/nanoclay (NC-CILE) nanocomposites, promoting the electron transfer and electrocatalytic activity. Considerable stability, electrocatalytic properties, and large surface area make it possible to exploit low detection limit sensors for long term applications in real samples and wide linear dynamic ranges. The electrochemical behavior of the prepared NC‑CILE electrode was diagnostically studied by cyclic voltammetry and differential pulse voltammetry analyses. The response currents were linear in the DP and UA concentration ranges of 1.0–100.0 and 0.1–10.0 μM, respectively. In addition, at 50.0 µM of AA, the detection limits for DP and UA were estimated to be around 0.45 and 0.025 μM (S/N = 3), respectively.