In this paper, soluble starch was studied as a depressant of hematite during flotation separation of apatite using sodium oleate as a collector. Surface charge measurement, soluble starch adsorptions, Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) were used to understand the interaction mechanisms between minerals (hematite and apatite) and soluble starch. The results indicated that chemical interaction between hematite and soluble starch was present, and supported the bonding of hydroxyl, while physical adsorption of soluble starch molecules with apatite occurred. Results of micro-flotation studies suggested that soluble starch was considered as a selective depressant for hematite. The maximum recovery difference between hematite and apatite of 77.5% was obtained with 40 mg/dm3 soluble starch. The flotation experiment results of natural iron ore showed that flotation indexes with 59.73% Fe, iron recovery of 81.5% and 75.68% of dephosphorization ratio were achieved at a soluble starch dosage of 60 mg/dm3. However, a higher dosage of soluble starch addition caused the difficulty for flotation separation of apatite from hematite. Our results provided theoretical basis for the flotation separation of apatite from iron oxide ores.