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The effect of polyether on the separation of pentlandite from serpentine has been studied. In addition to flotation and sedimentation tests, electrophoresis and adsorption tests have been conducted. The flotation and sedimentation results show that serpentine impairs flotation performance of pentlandite, by adhering to the pentlandite particles. Addition of the polyether could promote the dispersion of the mixed sample of pentlandite and serpentine in alkaline conditions and significantly reduce adverse effects of serpentine on the pentlandite flotation. The electrophoresis and adsorption tests show that polyether can selectively adsorb onto pentlandite surface through hydrophobic reaction and remove serpentine slime particles from pentlandite surfaces by steric hindrance effect.