The Holographic Particle Velocimetry system can be a promising optical tool leer the measurements of three dimensional particle velocities. In this research, validation experiments for the development of holographic particle velocimetry system for spray droplets were conducted with measuring the velocities of glass beads on a rotating disk. Uncertainty analysis was performed to identify the sources of all relevant errors and to evaluate their magnitude. The measurement results of distance between glass beads, size, and velocities obtained with holographic method are compared reasonably well with the known values within acceptable range of errors.