A novel method for studying the ionization step of matrix-assisted laser desorption/ionization (MALDI) is demonstrated. A 193-nm pulse from an ArF excimer laser is used to photodissociate a portion of a plume of insulin ions generated by MALDI. Laser photodissociation (LPD) creates a "hole", i.e., a negative spike in the insulin peak in the time-of-flight (TOF) mass spectrum. The position of the hole in the mass spectrum provides useful measurements of the characteristics (position, time, and velocity) of insulin ions shortly after their creation. Although the performance of the method can be further improved, the data obtained could be used to refine our current understanding of MALDI and to improve the resolution of MALDI-TOFMS.