Polymyxin　B　(PMB)　was　an　antibiotic　with　highly　effective　antibacterial　effect　but　narrow　safety　interval,　and　its　residues　in　food　had　attracted　widespread　attention.　It　was　important　to　develop　an　accurate　method　for　the　rapid　detection　of　PMB　in　animal　foods.　In　this　work,　we　had　established　a　ratiometric　sensing　system　based　on　the　formation　of　supramolecular　assemblies　of　pyrenyl　probes,　which　were　driven　by　the　synergy　of　noncovalent　forces　such　as　multiple-electrostatic　and　7C-7C　stacking　interactions.　Compared　with　the　traditional　fluorescence　detection　based　on　the　single　wavelength　change,　the　present　approach　showing　two-wavelength　fluorescence　response　could　reduce　the　interference　of　other　factors　making　the　experimental　results　more　accurate.　The　sensor　exhibited　high　sensitivity　and　selectivity　with　a　low　detection　limit　(28.3　nM).　This　method　could　be　used　to　realize　visual　detection　and　had　a　visual　detection　limit　of　1　mu　M.　As　we　had　learned　yet,　this　was　the　first　ratiometric　sensor　for　PMB　detection　in　aqueous　solution.　We　believed　all　our　preliminary　would　not　only　provide　a　complementary　strategy　for　the　detection　of　PMB,　but　also　develop　some　new　ideas　for　the　construction　of　sensors　for　rapid　antibiotic　detection.