In this study, the morphological features and the chemical composition characteristics of Opuntia ficus-indica and Opuntia humifusa fruits were investigated. The length of minor and major axis, and weight of O. ficus-indica and O. humifusa fruits were compared as the morphological features. The characteristics of components such as color, moisture, crude protein, ash, fat, dietary fiber, mineral, and amino acids were evaluated. The values of O. ficus-indica fruits were higher than those of O. humifusa fruits in each morphological feature such as minor axis, major axis, and weight. According to the Hunter color index results, O. humifusa fruits showed higher values of L (lightness) and b (yellowness), whereas O. ficus-indica fruits showed higher value of a (redness). The ΔE value between two samples was 19.80. The O. ficus-indica fruits had higher carbohydrate content and lower content of crude ash than those of O. humifusa fruits. Both samples showed very high dietary fiber contents, and the major minerals were K and Ca. Glutamic acid was the major amino acids in both samples. In free amino acids contents, O. ficus-indica had higher tyrosine and lower glutamic acid level than those of O. humifusa fruits. γ-aminobutyric acid was found in both samples, whereas taurine was found in O. ficus-indica fruits only. Taken together, the morphological features and chemical quality of O. ficus-indica and O. humifusa fruits showed difference although both of them were originated from the same genus.