Abstract: In order to improve the efficiency of fishbone calcium extraction, the high intensity pulsed electric fields (PEF) technique was used for assisted hydrolyzing the fishbone. The single factor experiment and second general revolving combination design results show that the pulse number, the electrical field strength, the mass ratio of citric acid and malic acid are the significant factors on the extraction efficiency of fishbone. With analysis of the regression mathematical model of extraction efficiency and the impact factor; the optimal combination of parameters was found that pulse number was 8, electrical field strength was 25 kV/cm, mass ratio of citric acid and malic acid in mix-acid was 1:1 g/g, acid charge proportion was 1:1 g/g, and water-powder ratio was 12:1 mL/g. Under the optimized condition, the extraction rate of 4 g fishbone power hydrolysate was up to 84.2%. The results show that comparing with ultrasonic technique, high intensity pulse electric fields is less time-consuming with higher extraction effeciency. The study can provide a reference for further studies of calcium fishbone extraction by PEF technique and the development of high calcium fishbone oral liquid.