Degradation products of V-type nerve agents are important markers of these toxic chemical warfare agents, hence their detection and identification is of high importance from verification point of view of Chemical Weapons Convention (CWC). We focused on detailed investigation of fragmentation of N, N-dialkylaminoethanesulfonic acids, degradation products of V-type nerve agents using liquid chromatography-tandem mass spectrometry (LC−MS/MS), a LCQ Deca XP max ion trap mass spectrometer via an electrospray ionization in ESI mode. Capillary and tube lens voltages were optimized to give maximum response to m/z 210 [M + H]+ ion of N,N-diisopropylaminoethanesulfonic acid. The main fragmentation pathways are loss of alkyl groups attached to the nitrogen atoms. Within the aims of CWC, LC−ESI-MSn represents an important tool to control the production of prohibited compounds among others due to a possibility of easy monitoring of their content in environmental matrices. The knowledge of the fragmentation pathway will be useful for the unambiguous identification of degradation products of V-type nerve agents in human urine matrices as well as environmental matrices in the Organization for the Prohibition of Chemical Weapons proficiency test.