This article describes the highly sensitive and selective determination of epinephrine (EP) using self-assembled monomolecular film (SAMF) of 1,8,15,22-tetraamino-phthalocyanatonickel(II) (4a-Ni II TAPc) on Au electrode. The 4a-Ni II TAPc SAMF modified electrode was prepared by spontaneous adsorption of 4a-Ni II TAPc from dimethylformamide solution. The modified electrode oxidizes EP at less over potential with enhanced current response in contrast to the bare Au electrode. The standard heterogeneous rate constant (k8) for the oxidation of EP at 4a-Ni II TAPc SAMF modified electrode was found to be 1.94 Â 10 À2 cm s À1 which was much higher than that at the bare Au electrode. Further, it was found that 4a-Ni II TAPc SAMF modified electrode separates the voltammetric signals of ascorbic acid (AA) and EP with a peak separation of 250 mV. Using amperometric method the lowest detection limit of 50 nM of EP was achieved at SAMF modified electrode. Simultaneous amperometric determination of AA and EP was also achieved at the SAMF modified electrode. Common physiological interferents such as uric acid, glucose, urea and NaCl do not interfere within the potential window of EP oxidation. The present 4a-Ni II TAPc SAMF modified electrode was also successfully applied to determine the concentration of EP in commercially available injection.