Ultrasonic contrast agents are used to assess perfusion conditions based on evaluation of the time-intensity curve. Such a curve reflects the concentration of microbubbles in the perfused area and the indicator-dilution theory is used to derive the volumetric flow rate from the measured concentration. Previous results have shown that the technique is not reliable in some conditions due to the shadowing effect. To overcome this problem, a contrast-specific technique using both the input and output time-intensity relationships is proposed; this contrasts with conventional techniques that utilize only the relationship directly from the perfused area. The proposed technique is referred to as the input-output time-intensity curve (IOTIC) method. In this work, the shadowing effect was studied experimentally and the efficacy of the IOTIC technique was assessed and compared with conventional techniques. The results indicate that the IOTIC technique eliminates the shadowing effect and provides a good correlation between the actual flow rate and measured flow-related parameters; thus, making quantitative estimation of perfusion feasible. Note that the IOTIC is applicable, based on the assumption that both the input and the output can be positioned within the same image plane; its clinical applications include situations where the perfused area cannot be effectively imaged by ultrasound (US). One example is the assessment of brain perfusion, and it will be used as a target clinical application of the IOTIC technique.