In this study, chitosan polymer was tested as a potential selective green depressant of pyrite in the bulk flotation of galena (PbS) and chalcopyrite (CuFeS2) from sphalerite (ZnS) and pyrite (FeS2) using sodium isopropyl xanthate as a collector and 4-methyl-2-pentanol (MIBC) as a frother. Flotation tests were carried out in a D12-Denver flotation laboratory cell in the presence and absence of chitosan and/or sodium cyanide depressant which is commercially used as pyrite depressant in sulfide mineral flotation process. Flotation recoveries and concentrate grades (assay) were studied as a function of polymer concentration and flotation time. It was found that at 50 g/ton, chitosan depressed 5.6% more pyrite as compared to conventional depressant NaCN at its optimum dosage. Furthermore, the measured assay values of pyrite in concentrates dropped by ~1.2% when NaCN depressant was replaced with chitosan polymer. Zeta potential measurements of galena, chalcopyrite, sphalerite, and pyrite suspensions before and after chitosan’s addition revealed that the polymer has preferential adsorption on pyrite minerals as compared to other sulfide minerals specially galena. Results obtained from this work show that chitosan polymer has a promising future as a biodegradable alternative to sodium cyanide for the purpose of depressing pyrite in sulfide minerals flotation.