For clinical biology, the ability to detect neurotransmitters in the human serum environment quickly, highly sensitively, and selectively is essential. Dopamine system dysfunction has been linked to a variety of nervous system diseases. As a result, its sensitive and selective detection is critical for the early diagnosis of diseases associated with excessive dopamine levels. So, platforms for sensitive and selective monitoring of dopamine concentrations are critically needed. In this study, we synthesized a facile flower-like nanocomposite material that is cost-effective and involves a rapid single-step synthesis of an Ag-Cu decorated ZnO nanoflower-like composite (Ag-Cu@ZnO-NFLC) with a high active surface area for comparison. Ag decorated ZnO (Ag@ZnO) and Cu decorated ZnO (Cu@ZnO) were synthesized and characterized by XRD, FTIR, and Raman spectroscopy analyses. By using methods such as cyclic voltammetry, differential pulse voltammetry (DPV), and amperometry, the electrochemical behavior of a GCE modified with a single-step synthesized Ag-Cu decorated ZnO nanoflower-like composite electrode material showed promising results with an ultra-low detection limit of 0.21 mu M and a high sensitivity of 0.68 mu A mM(-1) cm(-2) by employing differential pulse voltammetry (DPV). The dopamine sensor was also tested for current densities at different pH levels, and it exhibited good stability and reproducibility. Finally, the prepared sensor was used to monitor real-time human urine samples, yielding excellent results. The findings revealed that in actual sample analysis, the as-prepared sensor showed substantial promising results in detecting dopamine.