Our objective was to compare harmonic-to-fundamental frequency ratio peak (HFRp) analysis with conventional harmonic gray-scale densitometric analysis on the basis of ability to eliminate heterogeneity in ultrasound signals. Broadband radio frequency and harmonic data were obtained by using intermittent short-axis scans in 10 open-chest pigs before and during infusion of contrast microbubbles. HFRp and gray-scale intensity values were measured in 6 segments of left ventricular myocardium. In baseline images, the influence of anisotropy on HFRp values was significantly less than that in gray-scale intensities. In perfusion assessment with subtraction, contrast heterogeneity in HFRp values was significantly smaller than that in gray-scale intensities. The increase in HFRp values after subtraction was significantly greater than that in gray-scale intensities in lateral (P <.001), posterior (P <.0001), and inferior (P <.01) myocardium. HFRp analysis can compensate for baseline myocardial anisotropy and regional contrast heterogeneity. With background subtraction, HFRp analysis allows better quantification of myocardial perfusion.