Here, an ultrasensitive and low-cost electrochemical aptasensing assay is developed based on the applicability of a fabricated nanocomposite from nitrogen-doped graphene quantum dots (N-GQDs) and gold nanoparticles (AuNPs). A modified glassy carbon electrode (GCE) with the AuNPs@N-GQDs nanocomposite (AuNPs@N-GQDs/GCE) as an efficient platform has some unique properties such as high surface area and electrical conductivity. Furthermore, the prepared platform is capable of more loading of aptamer (Apt) molecules as a biological recognition element of Ibuprofen (IBP) on the modified electrode surface. It is noteworthy that in this study, riboflavin (RF) as a universal green probe is used for the first time for electrochemical detection of IBP. According to the proposed strategy and under the optimum condition, the unprecedented detection limit (LOD) of this assay (33.33 aM) is lower than previously reported analytical methods. The results demonstrate the ability of the nanocomposite for designing of the aptasensor, integrated within the electrode format, to cheaper and simpler detection of the IBP with a specificity and sensitivity sufficient for analysis in real samples. It seems that the proposed strategy based on the AuNPs@N-GQDs nanocomposite can be expanded to other nanomaterials. So, this is expected to have promising implications in the design of electrochemical sensors or biosensors for the detection of various targets.