A　pressurized　capillary　electrochromatography　(pCEC)　method　combined　with　an　online　concentration　was　developed　for　the　separation　and　determination　of　five　β2-blockers　(terbutaline,　salbutamol,　formoterol,　procaterol　and　salmeterol)　in　human　urine.　A　stearyl　methacrylate-based　monolithic　column　was　prepared　as　the　separation　column.　The　various　separation　parameters　including　acetontrile　concentration,　applied　voltage,　pH　and　concentration　of　the　running　buffer　were　investigated.　On-line　concentration　methods　in　combination　of　the　chromatographic　zone-sharpening　effect　and　field-enhanced　sample-stacking　effect　were　utilized　to　improve　detection　sensitivity.　The　on-line　concentration　parameters　including　injection　voltage,　injection　time,　as　well　as　sample　matrix　were　systematically　studied.　Compared　with　the　conventional　sample　injection,　the　on-line　concentration　technique　appeared　to　improve　the　corresponding　sensitivities　20{reversed　tilde}50-fold.　Furthermore,　good　precision　was　obtained　with　relative　standard　deviations　(RSDs)　for　retention　times　within　1.09%　and　for　peak　areas　less　than　3.55%　(n　=　5).　The　established　method　was　successfully　applied　to　the　determination　of　above-mentioned　β2-agonists　in　urine　samples.　The　recoveries　of　spiked　urine　samples　were　between　85.0　and　113.3%　with　RSDs　less　than　5.39%.　©　2015　WILEY-VCH　Verlag　GmbH　&　Co.　KGaA,　Weinheim.