We have designed three amphiphilic substituted urea compounds with varying chain lengths. These hydrophobic amphiphiles displayed aggregation-induced emission on solvent switching. The aggregates were further detailed by microscopy. The hydrophobicity of these AIEgens has been used to create water-repelling fluorescent surfaces. The AIEgen via the photoinduced electron transfer-mediated mechanism has been applied in the detection of nitroantibiotics. The analytical utility of the AIEgen is being demonstrated concerning the detection of nitroantibiotics in biofluids.