The sensitive monitoring of dopamine levels in the human body is of utmost importance since its abnormal levels can cause a variety of medical and behavioral problems. In this regard, we report the synthesis of nitrogen-doped graphene quantum dots (N-GQDs) from polyindole (PIN) via a facile single-step hydrothermal synthetic strategy that can act as an efficient electrochemical catalyst for the detection of dopamine (DA). The average diameter of N-GQDs was ∼5.2 nm and showed a C/N atomic ratio of ∼2.75%. These N-GQDs exhibit a cyan fluorescence color under irradiation from a 365 nm lamp, while PIN has no characteristic PL. The presence of richly N-doped graphitic lattices in the N-GQDs possibly accounts for the improved catalytic activity of N-GQDs/GCE towards electrocatalytic DA detection. Under optimum conditions, this novel N-GQDs-modified electrode exhibits superior selectivity and sensitivity. Moreover, it could detect as low as 0.15 nM of DA with a linear range of 0.001–1000 µM. In addition, the outstanding sensing attributes of the detector were extended to the real samples as well. Overall, our findings evidence that N-GQDs-based DA electrochemical sensors can be synthesized from PIN precursor and could act as promising EC sensors in medical diagnostic applications.