The high hydrophobicity and chemical inertness of poly(styrene‐divinylbenzene) (PS‐DVB) microspheres make their surface hydrophilic modification difficult. Here we describe a facile way to convert PS‐DVB microspheres to hydrophilic, then can be used as polar stationary phase for hydrophilic interaction chromatography. This approach utilizes the grafting of an acrylamide‐terminated lysine zwitterionic monomer onto PS‐DVB microspheres via free radical polymerization. The obtained stationary phase shows good hydrophilicity and a typical retention mechanism of hydrophilic interaction chromatography toward several model polar analytes. It also exhibits obvious zwitterionic properties and is capable of separating cationic and anionic analytes simultaneously. The column shows negligible bleeding level, much superior to silica‐based ones.