Amygdalin is widely distributed in loquat plant which belongs to Rosaceae and used as an important pharmaceutical compound. In this research, High Performance Liquid Chromatography (HPLC) was used to determine the content of amygdalin in different tissues of loquat fruit cv. ‘Dahongpao’, as well as leaves, roots and flowers. The results demonstrated that the kernel had the highest content of amygdalin among different fruit tissues, while the fruits flesh had the lowest content. Moreover, results showed significant differences in the content of amygdalin between kernel and leaves, while there were no significant differences between the content of kernel compared to roots and flowers. On the basis of above study, the content of amygdalin was determined in the kernel during different developmental stages and different cultivars. Results showed significant difference of the content of amygdalin in the kernel during different developmental stages and different cultivars. The results of this study can provide a theoretical basis for the comprehensive utilization of loquat fruit processing waste in the future.