The invention provides an experimental system and method for quantifying a preference hydraulic factor interval of fish. The experiment system comprises a water groove device and a data analysis system, wherein the water groove device comprises a water groove, a water inlet box and a tail water box which are arranged at two ends of the water groove and communicate with the water groove, and a rectification grid, a turbulence perturbator, a blocking net, a separation net, a tail door and a water circulation system which are arranged in the water groove; and the data analysis system comprises aCCD camera, a telescopic T-shaped support, a video recorder and a data processor. According to the invention, on the basis of an open water groove, a constant turbulence field is generated through theturbulence perturbator, and the preference hydraulic factor interval range of target experimental fish is obtained by analysis according to the real-time recorded upstream movement track change of the fish. The experimental system provided by the invention is reasonable in design and reliable in data for determining the preference hydraulic factor interval range of the fish, can provide help forconstruction of the hydrodynamic environment of a fish habitat and design and optimization of fishway hydraulic parameters; meanwhile, the water groove is convenient to be disassembled, assembled andtransported, and facilitates indoor experiments and field experiments.