We report the first direct evidence of detuning of stimulated Brillouin scattering (SBS) by a velocity gradient, which was achieved by directly measuring the frequency shift of the SBS-driven acoustic wave relative to the local resonant acoustic frequency. We show that in the expanding part of the plasma, ion-acoustic waves are driven off resonance which leads to the saturation of the SBS instability. These measurements are well reproduced by fluid simulations that include the measured flow.