Isoproterenol (ISO) is a synthetic amine that used to treat heart attacks, allergic emergencies, bronchitis, and cardiac chock. But on the other hand, excessive use of ISO can cause a significant decrease in blood pressure, as well as cause heart failure and/or cardiac arrhythmia, or in sometimes cause vasodilation of skeletal muscle arteries. Therefore, determination of ISO in various specimens is very important. The present attempt was made to introduce a sensitive electrochemical sensor by using carbon paste electrode (CPE) modified with multiwall carbon nanotubes/ZnCo-Zeolite imidazole frameworks and ionic liquid for determination of ISO at the presence of acetaminophen (AC). The characteristics of the as-fabricated sensor were explored via scanning electron microscope, Fourier transfer infra-red spectroscopy and X-ray powder diffraction. The electro-oxidation current of ISO onto the modified electrode surface was linear in the range of 0.04–580.0 μM and a limit of detection 9.8 nM was calculated according to 3Sb/m. For assesses the practical applicability of the modified electrode, the values of ISO and AC in serum and urine samples were determined using the calibration plot and as a result, appropriate relative standard deviations (2.6–3.5 %) and recoveries (97.2–103.9 %) were obtained.