In this work, a nanocomposite containing gold (Au) nanofibers decorated iron-metal–organic framework (Fe-MOF) was successfully synthesized for electrochemical detection of acetaminophen (AAP). The as-synthesized Au@Fe-MOF nanocomposite was confirmed by various characterization techniques. Morphological analysis showed that the Au nanofibers with an average size of less than 10 nm were dispersed on the Fe-MOF. Cyclic voltammetric analysis showed that the Au@Fe-MOF nanocomposite showed well-defined redox peaks with higher current than that of GCE and Fe-MOF. The Au@Fe-MOF/ GCE exhibited a linear range, sensitivity, and detection limit of 0.5–18 μM, 4.95 μM/μA/cm2, and 0.12 μM, respectively. The Au@Fe-MOF/GCE showed a very low response for the interference materials. The real sample analysis revealed that the Au@Fe-MOF/GCE showed good recovery towards the AAP in urine and paracetamol. Therefore, the developed sensor can be used for quality control of AAP.