Reverse flotation is an effective method for separating apatite from gangue minerals (such as dolomite) in collophanite ore. However, collophanite ore flotation using traditional fatty acid collectors is uneconomical because of the relatively high cost of the raw materials. In this work, saponified gutter oil fatty acid (GOFA) was used as the collector, and its effect on collophanite ore flotation and the mechanism of adsorption were studied. The results showed that Ca2+ is the active site for reaction with GOFA and initiates apatite flotation. The dissolution of Ca2+ from dolomite was found to be thermodynamically more favorable than that from apatite, and the relative abundance of Ca2+ enhanced the reaction of dolomite with GOFA. XPS analysis revealed that the interaction between GOFA and magnesium on the dolomite surface strengthened chemisorption. As magnesium is a major constituent of dolomite, but not of apatite, GOFA was selectively chemisorbed onto the dolomite surface. Closed-circuit reverse flotation tests using GOFA as the collector resulted in an excellent concentrate with 28.46wt% P2O5 and 0.65wt% MgO, corresponding to a P2O5 recovery of 87.2%. GOFA was demonstrated to exhibit outstanding selectivity for dolomite in the reverse flotation of collophanite ore. The use of GOFA as an environment-friendly, cost-effective, and selective collector is expected to provide new insights into low-cost collophanite ore flotation.