A monolithic column with the structure of three-dimensional bicontinuous skeleton was prepared by in-situ atom transfer radical polymerization（ATRP） with ethylene glycol dimethacrylate（EDMA） as monomer,methanol and hexane as binary porogenic agents,ethy bromopropionate（EBP） as crosslinker and copper bromide and N，N，N′，N′，N″-pentamethyldiethylenetriamine as the catalystical system.Effects of ratio of crosslinker and catalyst on the structure of the monolithic column were studied in details.The surface and micron through-pore and micro skeleton structure of the monolithic column were characterized by scanning electron microscope（SEM） and mercury intrusion method.Then the monolithic column was further modified through grafting copolymerization of butyl methacrylate initiated by —Br group on the surface of monolith.The modified monolithic column with hydrophobic surface was successfully applied in the separation of three steroid drugs（e.g.hydrocortisone,testosterone and progesterone） and four proteins（e.g.cytochrome C,myoglobin,ribonuclease A and ovalbumin） by reversed phase high performance liquid chromatography（RP-HPLC）.