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In this research, a novel collector cetyl trimethyl ammonium chloride (CTAC) was used to separate hematite from quartz via reverse flotation for the first time. Micro-flotation tests showed that CTAC had a strong ability to selectively collect quartz and that a separation of hematite from quartz could be accomplished with a concentration of 0.00263 mmol/L CTAC. Zeta-potential measurements indicated that the positive CTAC⁺ species could selectively increase the surface potential of quartz, but that it had rather a weak effect on the hematite. X-ray photoelectron spectroscopy (XPS) detection indicated that CTAC had a stronger binding affinity to oxygen sites on the surface of quartz than on hematite, resulting in a large amount of CTAC being predominantly adsorbed onto quartz. This was supported by the atomic concentration of C1s and N1s of quartz after CTAC treatments were 4.25 and 2.84 times higher than hematite, respectively.