A novel styrene-divinylbenzene copolymer monolithic capillary column was prepared using dimethylsulfoxide/dodecanol as binary porogenic agent.The polymer was prepared with styrene as monomer,divinylbenzene as crosslinker,benzoperoxide as initiator.In order to optimize the properties,the contents of the polymerized mixtures were investigated.Key variables such as composition of porogenic agent and crosslinking degree and content of porogenic agent in the mixtures could be controlled easily during the on-column polymerization.The PS-DVB monolithic column was characterized by scan electric microscopy(SEM),inverse size exclusion chromatography(ISEC),and measured under back pressure at different flow rates to evaluate the porous properties.Homologous compounds of benzene and some proteins were used as objective probes in reverse phase chromatography mode for evaluating the separation performance of the synthesized column.The optimum preparation conditions were as follows:the volume ratio of styrene and divinylbenzene and dimethylsulfoxide and dodecanol was 18.7∶15.3∶13.2∶52.8,that is,the content of dimethylsulfoxide in binary porogenic agent occupied 20%,the degree of crosslinking tooked up to 45% and the content of binary porogenic agent reached to 66%.The flow rate reached to 104 μL?min-1 in the experiment for the separation of protein,and the line speed was about 12 mm?s-1,while the conventional speed was about 1-2 mm?s-1.With the advantage of excellent chromatographic separation efficiency for biological macromolecules,the monolithic capillary column could be available in other separation modes if it is chemically modified.