【Objective】In order to study sugarcane chloroplasts proteomics， the methods for isolation and purification of intact chloroplast and its protein extraction from leaves of sugarcane seedling were compared. 【Method】The leaves of young seedling of sugarcane variety ROC22 were sampled to isolate the chloroplasts by differential centrifugation. The obtained intact chloroplasts were isolated and purified using sucrose density gradient centrifugation and Percoll gradient centrifugation. The proteins in chloroplast were extracted and analyzed by SDS-PAGE electrophoretic. 【Result】The results indicated that both density gradient centrifugation methods could obtain high rate and high purity of intact chloroplasts， but sucrose density gradient centrifugation saved the time and cost， and resulted in high purity and integrity of chloroplasts. The SDS-PAGE profiles of proteins， separated from the intact chloroplasts， were rich and clear， with high quantity and quality of proteins.【Conclusion】Compared with Percoll gradient centrifugation method， sucrose density gradient centrifugation method was suitable for proteomics studies of sugarcane at subcellular level.