In this paper, monodisperse polystyrene microspheres were prepared in a dispersion polymerization process. Then polystyrene microspheres were used as seeds, butyl phthalate as swelling agent, styrene as monomer, divinylbenzene as crosslinker, toluene as porogen, by using the method of seed swelling polymerization successfully made a narrow particle size distribution of porous high crosslinked polystyrene - divinylbenzene microspheres (PS - DVB). And porous microspheres were used as reversed-phase chromatographic packing matrix, and study the influence of the amount of crosslinker and porogen on the morphology of microspheres, particle size and pore structure parameters. The results show that the microspheres ball roundness, narrow particle size distribution, controllable pore structure, suitable for high performance liquid chromatography (HPLC) packings.