The traditional separation process of pyrite and marmatite is carried out under highly alkaline conditions. Therefore, a large amount of lime is demanded and the zinc recovery cannot be guaranteed. However, under weakly alkaline conditions, copper-activated pyrite has good floatability, which is difficult to separate from marmatite. In this paper, ammonium chloride (NH4Cl) is used for depressing the flotation of copper-activated pyrite to achieve the separation of these two minerals under weakly alkaline environment. The flotation tests show that NH4Cl can significantly reduce the floatability of pyrite in weakly alkaline conditions. The results of adsorption tests and X-ray photoelectron spectroscopy (XPS) analyses indicate that NH4Cl can obviously change the composition of pyrite surface by increasing the content of iron/copper hydroxide and reducing the content of copper sulfides. Calculation of the solution composition demonstrates that the addition of NH4Cl results in the occurrence of Cu(NH3)n2+ and the pH buffering property. Based on these results, it can be concluded that the depression of NH4Cl on copper activated pyrite is mainly derived from two aspects: 1) the pH buffering property of the conjugated acid-base pair (NH4+/NH3) can impede the decline of OH- concentration, which results in more hydroxide adsorbed on pyrite; 2) NH3(aq) competes with the pyrite surface to consume Cu2+ through complexation, which causes a reduction in the amount of copper sulfides formed on the pyrite surface.