Samples of flowers of wild Lavandula stoechas L. spp. stoechas populations were collected in three areas of Sicily (Italy) and were characterized in agronomic and chemical terms. Essential oil (EO) was extracted by hydrodistillation and analyzed by GC-FID and GC-MS. GC-FID and GC-MS analyses permitted identification of 89 compounds from the EO. The samples were separated into 3 groups using PCA (Principal Component Analysis) statistical method, with reference to the chemical composition of the EO. All three Sicilian populations of lavender were identified as the fenchone chemotype with percentage content ranged between 52.8-71.1%. The population of Partinico showed the highest dry weight of flowers per plant (221.3 g), but the lowest EO yield (0.37%). The essential oils of the three wild Sicilian populations of L. stoechas L. spp. stoechas showed a greater chemical differentiation than those obtained from other Mediterranean areas.