Both E-Nose and HS-SPME-GC-MS techniques were applied to identify volatile components from gonad of male Eriocheir sinensis cultured with four different eco-environment modes (pond, rice-field, lake, and Yangtze River). The results of E-Nose showed that the flavor profiles of gonad in male Eriocheir sinensis cultured in lake and rice-field were similar, while the flavor profiles of gonad from pond and Yangtze River were unique and could be distinguished well. GC-MS was used to analyze the type and content of volatile components in male gonad, and the main aroma compounds were selected according to their thresholds (OAV≥1). The results showed that the volatile components in the gonad of lake and rice-field male crab were similar in composition and content, but in the gonad of pond and Yangtze male crab, the composition of volatile components was more complicated, and the contents of hexanal (634.36 ng/g), benzaldehyde (1361.98 ng/g), and 2-pentylfuran (231.11 ng/g) in the gonad of male crab from the Yangtze River were significantly higher than those in the other three male gonad. In conclusion, this study showed that the flavors of gonad in male Eriocheir sinensis cultured in lake and rice-field were similar, while the flavors of gonad from pond and Yangtze River were unique, and among the four different modes, the gonad of Yangtze River has better flavor than others.