In this paper, the effect of sodium alginate (SA) on the flotation separation of molybdenite (MoS 2) from chalcopyrite using kerosene as collector was systematically investigated. The results of single-mineral micro-flotation tests indicated that SA exhibited strong depression on chalcopyrite flotation while it imposed no impact on the floatability of molybdenite. However, in the chalcopyrite-molybdenite mixed-mineral flotation system, the presence of chalcopyrite significantly increased the depressing effect of SA on molybdenite flotation, leading to a considerable reduction in the flotation selectivity. The negative impact of chalcopyrite on the performance of SA in molybdenite flotation was eliminated by adding a certain dosage of kerosene prior to SA. A concentrate containing 53.43% of molybdenum (Mo) was obtained at 76.90% of recovery using 19 mg/L kerosene and 40 mg/L SA at pH 5.4. Zeta potential measurements indicated that the adsorption of SA on chalcopyrite surfaces was stronger than that on molybdenite surfaces, which agreed with the single-mineral flotation test results. The adsorption of SA on chalcopyrite was further confirmed to be chemisorption by Fourier-transform infrared spectroscopy (FTIR) spectra analyses. When Cu 2+ appeared in solution, the flotation of molybdenite was strongly depressed by SA. Mechanism analyses indicated that more active sites were generated on molybdenite surfaces after the addition of Cu 2+ , thus promoting the adsorption of SA.