In this study, the properties of (CH3)2CHOC(S)NHC2H5(Dow: Z-200) after the ultrasonic pretreatment was characterized by employing surface tension, viscosity, and Fourier transform infrared (FTIR) spectroscopy, and its influence on chalcopyrite rough selecting was investigated. The results indicate that the pretreated Z-200 can improve the index of chalcopyrite roughing. And, under the same reagent system, the recovery of copper reached 82.84% which was an increase of 24.44% compared with the untreated when Z-200 after ultrasonic pretreatment was applied to the rough separation of chalcopyrite. The reason why ultrasonic can strengthen the flotation effect of Z-200 on chalcopyrite is that ultrasonic pretreatment can decrease the surface tension and viscosity of Z-200 and enhance its foaming performance. Meanwhile, the ultrasonic cavitation destroys the molecular structure of Z-200, so that the relative proportion of methyl absorption peak and amine absorption peak in Z-200 increases. This also further improves the collection performance and foaming performance of Z-200 and strengthens its separation effect on chalcopyrite. The research provides a new idea for Z-200 to act on chalcopyrite and improve its flotation efficiency, reduce the amount of flotation reagent, and its pollution to the environment. It also provides a theoretical basis for expanding the application of ultrasonic technology in the field of flotation.