MALDI-TOF MS profiling has proved to be efficient for arthropod identification at the species level. However, prior to entomological monitoring, the reference spectra database should cover relevant species. Here, 74 specimens were field-collected from 11 mosquito species captured in two distinct European areas and used either to increment our database or for blind tests. Misidentification was not noted, underlining the power of this approach. Nevertheless, three out of the 26 specimens used for the blind test did not reach the significant identification threshold value set, attributed to lower spectral quality. In the future, the quality control spectra parameters need to be defined to avoid not achieving significant threshold identification.