The interfacial diffusive contaminant (phosphorus) release from permeable sediment layer into overlying water column under a unidirectional unsteady (periodic) flow condition was experimentally measured and analyzed. The experimental results indicate that the gross diffusive contaminant release rate is substantially enhanced as compared to that under a steady flow condition, and this enhancement trend is much more pronounced in an immediate release stage. The interfacial diffusive contaminant release rate tends to increase with the increasing flow velocity, decreasing period and augmenting amplitude for the case of the unsteady flow. The additional interfacial diffusive contaminant release under the unsteady flow condition may be related to the hydrodynamic response of the diffusive boundary layer to the flow unsteadiness of the overlying water, depending upon not only the periodic thickness variation of the diffusive boundary layer immediately above the sediment-water interface modulated by the temporal flow velocity of the overlying water column but also the intensified turbulent mixing between the overlying water and the pore-water within the superficial sediment layer induced by an alternate acceleration/deceleration fluctuation during each period.