The effects of various pulsed electric field (PEF) parameters on the extraction of polyacetylenes from carrot slices were investigated. Optimised conditions with regard to electric field strength (1–4 kV/cm), number of pulses (100–1500), pulse frequency (10–200 Hz) and pulse width (10–30 μs) were identified using response surface methodology (RSM) to maximise the extraction of falcarinol (FaOH), falcarindiol (FaDOH) and falcarindiol-3-acetate (FaDOAc) from carrot slices. Data obtained from RSM and experiments fitted significantly (p < 0.0001) the proposed second-order response functions with high regression coefficients (R2 ) ranging from 0.82 to 0.75. Maximal FaOH (188%), FaDOH (164.9%) and FaDOAc (166.8%) levels relative to untreated samples were obtained from carrot slices after applying PEF treatments at 4 kV/cm with 100 number of pulses of 10 μs at 10 Hz. The predicted values from the developed quadratic polynomial equation were in close agreement with the actual experimental values with low average mean deviations (E%) ranging from 0.68% to 3.58%.