For many tasks in failure and damage analysis of surfaces deteriorated in heavy tribological contact, the detailed characterization of used lubricants and their additives is essential. The objective of the presented work is to establish accessibility of tribostressed surfaces for direct characterization via modified atmospheric pressure-matrix assisted laser desorption/ionization-mass spectrometry (m-AP-MALDI-MS). Special target holders were constructed to allow target samples of differing shape and form to fit into the desorption/ionization chamber. The best results of desorption and ionization on different target materials and varying roughnesses were achieved on smooth surfaces with low matrix/substrate interaction. M-AP-MALDI characterization of tribologically stressed steel surfaces after pin-on-disc sliding wear tests (SRV-tribotests) yielded positive identification of used friction modifier additives. Further structure elucidation by electrospray ionization mass spectrometry (ESI-MS) and measurements of worn surfaces by time-of-flight-secondary ion mass spectrometry (TOF-SIMS) accompanied findings about additive behavior and deterioration during tribological contact. Using m-AP-MALDI for direct offline examinations of worn surfaces may set up a quick method for determination of additives used for lubrication and general characterization of a tribological system.