The effects of high-intensity pulsed electric field (HIPEF) treatment variables (frequency, pulse width, and pulse polarity) on the lycopene, vitamin C, and antioxidant capacities of tomato juice were evaluated using a response surface methodology. An optimization of the HIPEF treatment conditions was carried out to obtain tomato juice with the highest content of bioactive compounds possible. Samples were subjected to an electric field intensity set at 35 kV/cm for 1000 micros using squared wave pulses, frequencies from 50 to 250 Hz, and a pulse width from 1 to 7 micros, in monopolar or bipolar mode. Data significantly fit (P < 0.001) the proposed second-order response functions. Pulse frequency, width, and polarity significantly affected the lycopene, vitamin C, and antioxidant capacities of HIPEF-treated tomato juice. Maximal relative lycopene content (131.8%), vitamin C content (90.2%), and antioxidant capacity retention (89.4%) were attained with HIPEF treatments of a 1 micros pulse duration applied at 250 Hz in bipolar mode. Therefore, the application of HIPEF may be appropriate to achieve nutritious tomato juice.