To study the number of ethylene oxide (EO) groups effect on the flotation of rutile, three collectors with different EO number were evaluated: AC1203 (with three EO number), AC1210 (with ten EO number), AC1215 (with fifteen EO number). In addition to the flotation experiments, zeta potential measurements, collector adsorption experiments and XPS analysis were conducted to elucidate the adsorption mechanism. The results of flotation demonstrated that dodecylamine polyoxyethylene ether exhibited a profound collecting ability towards rutile mineral, it was worth mentioned that the recovery of rutile decreased with the increase of EO number. On the basis of zeta potential tests and XPS analysis, both protonated and neutral tertiary amine groups could act with rutile surface through electrostatic effect and hydrogen bond interaction. Furthermore, the results of collector adsorption experiments showed that with the increase of EO number, a lower adsorption density of collectors on rutile surface could be caused due to the steric hinder effects.