The use of supercritical carbon dioxide (Sc-CO2) extraction at 10 and 14 MPa and 40 °C and on-line fractionation using two separators (Sep 1: 7 MPa/25 °C; Sep2: 5 MPa/15 °C) to recovery volatile compounds from the inflorescences of fiber type Cannabis sativa L. was investigated by HS-SPME/GC–MS and direct GC–MS and compared with hydrodistillation. The best results were obtained by Sc-CO2 extraction carried out at 10 MPa and 40 °C. Under these operating conditions, cuticular waxes covering the surface of flowers were collected in the first separator and volatile compounds (100%) in the second. The superior quality of this last extract was proved by the perfect overlapping of its HS-SPME/GC–MS volatile profile to that of inflorescences. The recovery of fractions with different composition and biological properties, made the inflorescences of fiber type Cannabis sativa L suitable for cosmetic and/or food industry.