This study investigates the efficacy of high-voltage electrical field (PEF) treatment for sterilizing sugarcane juice. The system comprises a direct current (DC) pulsed high-voltage power supply, a treatment chamber, and a fluid flow system. Using Escherichia coli and Candida albicans as test organisms, we explored the impact of various voltage levels (10, 20, 30, and 40 kV) and flow rates (1, 2, 3, and 4 L/min) over 60-minute intervals. Results indicate that E.coli was effectively inactivated, with higher voltage levels leading to greater microbial reduction, particularly at 30 and 40 kV. Conversely, C. albicans exhibited lower susceptibility, possibly due to its robust cell wall. Adjustments in flow rate revealed that higher voltages could mitigate the negative effects of increased flow rates, achieving optimal sterilization. Sweetness (°Brix) and juice temperature remained relatively stable, demonstrating that PEF treatment preserves juice quality better than traditional thermal pasteurization.