To assess whether phantomless calcium-hydroxyapatite (HA) specific bone mineral density (BMD) measurements with dual-layer spectral computed tomography are accurate in phantoms and vertebral specimens. Ex-vivo human vertebrae (n = 13) and a phantom containing different known HA concentrations were placed in a semi-anthropomorphic abdomen phantom with different extension rings simulating different degrees of obesity. Phantomless dual-layer spectral CT was performed at different tube current settings (500, 250, 125 and 50 mAs). HA-specific BMD was derived from spectral-based virtual monoenergetic images at 50 keV and 200 keV. Values were compared to the HA concentrations of the phantoms and conventional qCT measurements using a reference phantom, respectively. Above 125 mAs, errors for phantom measurements ranged between -1.3% to 4.8%, based on spectral information. In vertebral specimens, high correlations were found between BMD values assessed with spectral CT and conventional qCT (r ranging between 0.96 and 0.99; p < 0.001 for all) with different extension rings, and a high agreement was found in Bland Altman plots. Different degrees of obesity did not have a significant influence on measurements (P > 0.05 for all). These results suggest a high validity of HA-specific BMD measurements based on dual-layer spectral CT examinations in setups simulating different degrees of obesity without the need for a reference phantom, thus demonstrating their feasibility in clinical routine.