In this study, bowl‐shaped N‐doped hollow carbon sphere‐containing mesoporous nanomaterials (BNHC) were used for the ultrasensitive electrochemical detection of isoprenaline (ISOP). The linear ranges of the BNHC‐based sensor were as follows: 0.05 – 15 μM; 15 – 70 μM, with the limit of detection (LOD) of 6.8 nM. And the BNHC‐based electrochemical sensor showed a good current response to ISOP and maintained 95.6 % of the initial current after 30 days of storage. Notably, when the BNHC‐modified sensor was used to detect the actual ISOP injection and urine samples, the sensor exhibited satisfactory recovery.