The　widespread　use　of　phenylurea　herbicides　(PHs)　is　a　serious　threat　to　human　health　and　environmental　safety.　These　pollutants　usually　have　similar　molecular　structures　but　distinct　toxicities.　Therefore,　developing　rapid　detection　methods　to　analyze　PHs　quantitatively　and　qualitatively　is　highly　desirable.　This　study　fabricated　a　microfluidic　glass　liquid　chromatography　(LC)　coupled　with　electrochemical　detection　(ECD)　and　Surface-Enhanced　Raman　spectroscopy　(SERS)　synchronous　system,　which　demonstrated　excellent　separation　and　analysis　performance　for　PHs.　The　complete　separation　of　three　PHs　could　be　achieved　using　the　microfluidic　chip　LC　with　a　theoretical　plate　number　of　342525　plates　m–1.　The　limit　of　detection　(LOD)　of　the　PHs　for　optimized　electrochemical　analysis　reached　0.0099–0.1388　mmol/L.　More　importantly,　the　detection　system　integrated　with　ECD　and　SERS　had　high　sensitivity　and　molecular　recognition　ability,　with　intra　and　inter-day　precision　of　less　than　6.7630%　and　7.5601%,　respectively.　The　recovery　of　the　PHs　could　reach　102.9861%　when　this　detection　system　was　applied　to　the　analysis　of　soil　extracts　in　the　small　potted　plant　and　actual　water　samples　in　the　environment.　It　is　expected　that　the　microfluidic　chip　with　the　LC-ECD-SERS　synchronous　detection　system　has　significant　potential　in　various　environmental,　food　safety,　drug,　and　biomedical　analyses.　©　2024　Elsevier　B.V.