Triclocarban　(TCC)　is　a　common　antimicrobial　agent　that　has　been　widely　used　in　medical　care.　Given　the　close　association　between　TCC　treatment　and　metabolic　disorders,　we　assessed　whether　long-term　treatment　to　TCC　at　a　human-relevant　concentration　could　induce　nephrotoxicity　by　disrupting　the　metabolic　levels　in　a　mouse　model.　Matrix-assisted　laser　desorption/ionization　mass　spectrometry　imaging　(MALDI-MSI)　was　applied　to　investigate　the　alterations　in　the　spatial　distributions　and　abundances　of　TCC,　endogenous　and　exogenous　metabolites　in　the　kidney　after　TCC　treatment.　The　results　showed　that　TCC　treatment　induced　the　changes　in　the　organ　weight,　organ　coefficient　and　histopathology　of　the　mouse　kidney.　MSI　data　revealed　that　TCC　accumulated　in　all　regions　of　the　kidney,　while　its　five　metabolites　mainly　distributed　in　the　cortex　regions.　The　abundances　of　79　biomolecules　associated　with　pathways　of　leukotriene　E4　metabolism,　biosynthesis　and　degradation　of　glycerophospholipids　and　glycerolipids,　ceramide-to-sphingomyelin　signaling　were　significantly　altered　in　the　kidney　after　TCC　treatment.　These　biomolecules　showed　distinctive　distributions　in　the　kidney　and　displayed　a　favorable　spatial　correlation　with　the　pathological　damage.　This　work　offers　new　insights　into　the　related　mechanisms　of　TCC-induced　nephrotocicity　and　exhibits　the　potential　of　MALDI-MSI-based　spatial　metabolomics　as　a　promising　approach　for　the　risk　assessment　of　agents　in　medical　care.　©　2024　The　Authors