A sensitive colorimetric and electrochemical sensor for measuring of epinephrine (EP) was developed based on CuO nanorods (NRs), and applicability of the sensor for detection of release epinephrine (EP) from living cells was evaluated. The CuO NRs was prepared using a facile and efficient method in low temperature and characterized by Scanning electron microscopy (SEM), Transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR) and Energy-dispersive X-ray spectroscopy (EDX). The CuO NRs exhibited laccase-like activity and could oxidize epinephrine (EP) to a colored product. No interference from the common interfering agents such as dopamine, ascorbic acid and uric acid was observed. Colorimetric sensor demonstrated a linear range of 0.6−18 μM with detection limit of 0.31 μM. Furthermore, the electrochemical study showed CuO NRs exhibited excellent electrocatalytic activity towards epinephrine oxidation. Differential pulse voltammetry signals increase with increasing of EP concentration in the range 0.04−14 μM, with a detection limit of 20 nM. Finally, the proposed sensor applied to perform real-time monitoring of epinephrine released by PC12 cells, indicating that CuO NRs provide a new platform for developing high-performance sensors in biological applications.