We analyzed localization of methyl 2-octynate (2-OAm) in sections of liver by nano particle assisted laser desorption/ionization (nano-PALDI) imaging mass spectrometry (IMS). 2-OAm was applied to mice skin and it permeated through the skin and accumulated in the liver. In livers of single dose mice, 2-OAm was delivered to the liver for 6 hours and excreted from the liver for 24 hours. On the other hand, in livers of long apply mice, 2-OAm was retained in the liver. Furthermore, we could be revealed that 2- OAm was accumulated in bile ducts by analyzing at a high resolution. In addition, CD8 staining indicated that an inflamed bile duct was observed. 2- OAm triggered the inflammation due to a coincident localization of 2-OAm and bile duct. This imaging approach is a promising technique for rapid quality evaluation of xenobiotics.