Purpose: Opuntia Humifusa has been used in the food and beauty industry after removing spines and glochids clearly. This study compared the methods used in removing spines and analyzed the design factors for developing a spine removal device. Method: This study compared the spine removal ratios in accordance with the length of brush, water spray pressure, the number of water spray, and the size of Opuntia Humifusa in a rotating brush device and a water spray device. In addition, this study compared the reversal ratios according to the inclination angle of a conveyor, the drop height of Opuntia Humifusa, and the speed of the conveyor to analyze the reversal factors. Results: The spines were not removed clearly in the rotating brush method, and the glochids were nailed deeply. The spine removal ratio was 96.9% with the water spray pressure of 20 $kgf/cm^2$ and the conveyor speed of 10 cm/s in the water spray method. The number of water spray was correlated with the spine removal ratio, and the average spine removal ratio was 95.6% with three cycles of water spray. The reversal ratio was 97% with the inclination angle of the conveyor $20^{circ}$, the drop height of 380 mm, and the conveyor speed of 10 cm/s. Conclusions: In order to develop a Opuntia humifusa spine removing device, this study compared the rotating brush and water spray methods. As a result, each spine removal performance of the rotating brush and water spray methods was 96.9% and 95.6%, respectively. Although the performance of the rotating brush method was slightly higher than that of the water spray method, the water spray method was suitable for removing spines from stem because the epidermis of stem was damaged and the glochids were nail deeply in the rotating brush method. Further studies on appropriate arrangement of spray nozzles, maintaining the optimal water spray pressure, the speed and angle control of the feeding conveyor, and devices for inducing the stem to the center will be needed in combining the water spray device and the reversal device.