The effect of ethyl thioglycolate organic small molecule inhibitors on chalcopyrite molybdenite flotation behaviour is investigated via single mineral micro-flotation tests, zeta potential tests, and X-ray photoelectron spectroscopy (XPS) analysis. Results of the flotation test indicate that ethyl thioglycolate organic small-molecule inhibitors can effectively separate Cu and Mo and selectively inhibit chalcopyrite under weak alkaline conditions. Infrared spectroscopy and XPS analysis show that hydrophilic functional groups C=O and -COOH in the ethyl thioglycolate organic small molecules can chemically adsorb onto the chalcopyrite surface. Moreover, ethyl thioglycolate has no obvious effect on zeta potential of molybdenite. Therefore, ethyl thioglycolate can effectively separate chalcopyrite and molybdenite.