Polycyclic aromatic hydrocarbons (PAHs) adsorbed on the surface of particulates collected in a working environment have been analyzed using fluorescence techniques. In particular, fluorescence measurements were carried out directly on the sampling filters, in solid phase, and for comparison, on the extract samples in solution. Fluorescence synchronous acquisitions allowed the detection of signals from different PAH compounds and, on the basis of the 0-0 transition energy, the assignment of the emission signals to chemical structures. In particular, the contribution of benzo-a-pyrene was easily detected and used to evaluate the sensitivity of the measurements, which is a few nanograms per milliliter, comparable with other analytical tools. The results of the fluorimetric investigation were validated through the comparison with the data obtained by gas chromatography (GC) analysis on the extracted samples, which allowed identification of the PAHs and the quantification of their distribution on the filters with different cut sizes. The agreement between the two series of data led to the conclusion that fluorimetric analysis directly on the sampling filter could be a new and cost-effective approach for the analysis of PAHs.