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In this study we discriminate three freshwater fish species (the shad Alosa agone Scopoli 1786, the whitefish Coregonus macrophthalmus Nüsslin1882 and the roach Rutilus rutilus Linnaeus 1758) by Matrix-Assisted Laser Desorption/Ionization- Time Of Flight Mass Spectrometry (MALDI-TOF MS) using both muscle and liver tissues. The technology enables to analyze tissues after a simple single-step extraction procedure without any further purification. The molecular profile of muscle tissues showed the most intense peaks at m/z range of 11,354.0 (±2.0 SD) Da, 3508.5 (±1.5 SD) Da and 8567.2 (±1.1 SD) Da for the shad, the whitefish and the roach respectively. The molecular profiles of liver tissues exhibit most of the highest peak intensities in the range between 2000 and 6000 m/z values. The roach shows the clearest pattern with high intensities detected at mass ranges between 3000 and 3550 Da with maxima at m/z 3035.2 (±0.2) Da and 3468.7 (±0.3) Da. The shad shows a shared high peak at m/z 3429.0 (±0.3) Da. The whitefish shows a group of major peaks in the m/z range of 3000-3700 Da with the highest being at 3635 (±0.3) Da. The overall signal pattern generated is highly specific for each species and, according to cluster analyses based on the total number of peaks, we could discriminate the three species.
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