Hydrophilic aromatic polyurethane (HAPU) microspheres were prepared through dispersion polymerization of a rosin-based polyurethane dispersion with C=C and styrene (St). The effects of the monomer ratio (i.e., waterborne rosin-based aromatic polyurethane (WRPU) to St), dispersant level, and reaction temperature on the properties of the microspheres were investigated; the effects of pH and adsorption temperature on the adsorption capacity of Orange II were also studied. The microspheres were characterized using Fourier transform infrared spectroscopy, energy-dispersive spectrometry, thermogravimetric analysis, laser particle size analysis, and scanning electron microscopy. The results showed that HAPU microspheres have been successfully synthesized and the produced microspheres exhibited good thermal stability and monodispersion. The optimum reaction conditions for the preparation of the microspheres were determined as a monomer ratio (m WRPU/m St) of 6:4 with 8 wt % poly(vinyl pyrrolidine) (on the basis of the mixed monomer) at 80 °C for 8 h. Under these conditions, the average particle size of the synthetic microspheres was 120 nm and the particle size distribution index was 0.442. The microspheres' adsorption capacity for Orange II reached 17.53 mg·g-1 when the solid-liquid ratio was 1 g·L-1, with an initial concentration of 100 mg·L-1 at pH 5, and the adsorption was conducted at 313 K for 3 h.