A　novel　capillary　electrochromatography　method　was　developed　by　using　a　polymethacrylate　ester　based　monolithic　column　for　the　simultaneous　separation　and　determination　of　five　nucleoside　and　purine　compounds　in　urine.　The　porous　monolithic　column　was　first　designed　by　mean　of　in　situ　copolymerizing　stearyl　methacrylate,　trihydroxymethylpropyl　trimethylacrylate,　and　2-acrylamido-2-methyl-1-propanesulfonic　acid　in　a　ternary　porogenic　solvent　including　cyclohexanol,　1,4-butanediol,　and　water.　The　performance　and　influence　factors　of　the　monolithic　columns　were　investigated　and　evaluated　by　a　CEC　method.　The　five　compounds　including　guanine,　adenine,　adenosine,　cytidine,　and　N6-methyladenosine　were　selected　as　analytes.　Under　the　optimized　condition　of　6.0　mM　phosphate　buffer　at　pH　5.0　without　ACN,　five　analytes　were　rapidly　separated　in　4　min　with　the　high　separation　efficiency.　The　recoveries　of　spiked　samples　were　ranged　from　89.0　to　109.0%　with　RSDs　less　than　4.83%.　Therefore,　the　proposed　method　could　possibly　provide　for　rapid　separation　and　detection　of　the　nucleoside　and　purine　compounds　in　biomedicine　sample.　©　2015　WILEY-VCH　Verlag　GmbH　&　Co.　KGaA,　Weinheim.