The accurate detection and quantification of Dopamine and Serotonin, two crucial neurotransmitters, play an essential role in understanding and treating a range of mental and neurological disorders. This research focused on the development of a novel method for the precise and straightforward identification of Dopamine and Serotonin, by integrating specific aptamers with gold nanoparticles (AuNPs). With the optimal conditions that were investigated, the standard curve for determining Dopamine and Serotonin was constructed with a high correlation coefficient (>
0.99), and the limits of detection (LOD) for Dopamine and Serotonin were 1.7 nM and 1.4 nM, respectively. Simultaneously, the selectivity of the method was evaluated in the presence of various interfering substances. The results indicated a relatively high selectivity of the method when interfering substances were present. These findings not only provided a robust foundation for the development of rapid, accurate, and sensitive analytical methods for Dopamine and Serotonin but also opened up new research directions in the field of healthcare and pharmaceuticals.