Spirulina (Arthrospira platensis) has been consumed by humans since ancient times. It is rich in high added-value compounds such as chlorophylls, carotenoids and polyphenols. Pulsed electric fields (PEF) is an innovative non-thermal technique that improves the extraction of bioactive compounds from diverse sources. PEF pre-treatment (3 kV/cm, 100 kJ/kg) combined with supplementary extraction with binary solvents at different times was evaluated to obtain the optimal conditions for extraction. In addition, the results obtained were compared with conventional treatment (without PEF pre-treatment and constant shaking) and different strains of Spirulina from diverse geographical locations. The optimal extraction conditions for recovering the bioactive compounds were obtained after applying PEF treatment combined with the binary mixture EtOH/H2O for 180 min. The recovery of total phenolic content (TPC) (19.76 ± 0.50 mg/g DM (dry matter) and carotenoids (0.50 ± 0.01 mg/g DM) was more efficient in the Spirulina from Spain. On the other hand, there was a higher recovery of chlorophylls in the Spirulina from China. The highest extraction of total antioxidant compounds was in Spirulina from Costa Rica. These results show that PEF, solvents and the condition of growing affect the extraction of antioxidant bioactive compounds from Spirulina. The combination of PEF and EtOH/H2O is a promising technology due to its environmental sustainability.