Porous polymer monolithic columns have been prepared by the direct free radical copolymerization of methacrylic acid and ethylene glycol dimethacrylate within the confines of a chromatographic column in the presence of toluene-dodecanol as a porogenic solvent. Recognition mechanism and effects of the chromatographic condition such as mobile phase composition, flow rate and temperature on the retention and separation were discussed. The results showed hydrogen-bonding interaction and hydrophobic interaction play an important role in retention and separation in this kind of monolithic column. Compared with traditional particle columns, the monolithic columns showed attractive significant interest because of their ease of preparation, high separation efficiency, low backpressure and fast analysis.