The purposes of this study were to assess the extent of the inflow effect on signal intensity (SI) for fast gradient-recalled-echo (GRE) sequences used to observe first-pass perfusion, and to develop and validate a correction method for this effect. A phantom experiment with a flow apparatus was performed to determine SI as a function of Gd-DTPA concentration for various velocities. Subsequently a flow-sensitive calibration method was developed, and validated on bolus injections into an open-circuit flow apparatus and in vivo. It is shown that calibration methods based on static phantoms are not appropriate for accurate signal-to-concentration conversion in images affected by high flow. The flow-corrected calibration method presented here can be used to improve the accuracy and robustness of the arterial input function (AIF) determination for tissue perfusion quantification using MRI and contrast media.