Electrochemical determination of synthetic opioids such as fentanyl is meaningful but still challenging no matter from a social or academic perspective. Herein, we report a series of novel electrocatalysts based on Fe-containing metal-organic frameworks and multi-walled carbon nanotubes (Fe-MOF/MWCNTs). The obtained Fe-MOF/MWCNT electrode materials all show ultrasensitivity on fentanyl determination. In particular, MOF-235/MWCNTs even exhibit an ultra-low limit of detection (LOD) of 0.03 μM with a wide linear range from 0.1 to 50 μM. Besides, this series of Fe-MOF/MWCNTs also displays excellent repeatability, selectivity, and stability. Moreover, they show good performance in real sample detection and achieve good recovery of 95.47%-102.41% and 96.62%-103.15% in blood and urine samples, respectively. This high performance in fentanyl determination is mainly contributed by the Fenton-like process and the adsorption function of the Fe-MOF. Therefore, these novel Fe-MOF/MWCNTs are promising electrocatalysts for point-of-care device fabrication and also have potential applications in fentanyl rapid test technology.