Ti–6Al–4V alloys undergo a multiple phase transformation sequence during electron beam powder bed fusion (EB-PBF) additive manufacturing, forming unique dislocation substructures. Thus, determining the dislocation density is crucial for comprehensively understanding the strengthening mechanisms and deformation behavior. This study performed time-of-flight neutron diffraction (TOF-ND) measurements of Ti–6Al–4V alloys prepared via EB-PBF and examined the dislocation density in the as-built and post-processed states using convolutional multiple whole profile (CMWP) fitting. The present TOF-ND/CMWP approach successfully determined the bulk-averaged dislocation density (6.8 × 1013 m−2) in the as-built state for the α-matrix, suggesting a non-negligible contribution of dislocation hardening. The obtained dislocation density values were comparable to those obtained by conventional and synchrotron X-ray diffraction (XRD) measurements, confirming the reliability of the analysis, and indicating that the dislocations in the α-matrix were homogeneously distributed throughout the as-built specimen. However, the negative and positive neutron scattering lengths of Ti and Al, respectively, lowered the diffraction intensity for the Ti−6Al−4V alloys, thereby decreasing the lower limit of the measurable dislocation density and making the analysis difficult.