Beetroot is a root vegetable rich in different bioactive components, such as vitamins, minerals, phenolics, carotenoids, nitrate, ascorbic acids, and betalains, that can have a positive effect on human health. The aim of this work was to study the influence of the pulsed electric field (PEF) at different electric field strengths (4.38 and 6.25 kV/cm), pulse number 10–30, and energy input 0–12.5 kJ/kg as a pretreatment method on the extraction of betalains from beetroot. The obtained results showed that the application of PEF pre-treatment significantly (p < 0.05) influenced the efficiency of extraction of bioactive compounds from beetroot. The highest increase in the content of betalain compounds in the red beet’s extract (betanin by 329%, vulgaxanthin by 244%, compared to the control sample), was noted for 20 pulses of electric field at 4.38 kV/cm of strength. Treatment of the plant material with a PEF also resulted in an increase in the electrical conductivity compared to the non-treated sample due to the increase in cell membrane permeability, which was associated with leakage of substances able to conduct electricity, including mineral salts, into the intercellular space.