Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) analysis of ionic liquid matrices (ILMs) prepared using pyridine and dihydroxybenzoic acid (DHB), such as 2,3-DHB and 2,5-DHB, displayed an unconven-tional peak at m/z 232.0, which was regarded as [DHB+pyridine-H] + . The peak at m/z 232.0 was not observed from other ILMs prepared using other DHB isomers, such as 2,4-DHB, 2,6-DHB, 3,4-DHB, and 3,5-DHB. Two requirements to observe the peak at m/z 232.0 in a DHB/pyridine ILM are suggested. First, carboxyl and hydroxyl groups must be located ortho to each other. Sec- ond, the secondary hydroxyl group must be located at a carbon with a high electron density. Based on these two requirements, a potential mechanism for the generation of the peak at m/z 232.0 is suggested.