Quantifying the correlation length of the tissue microstructure has shown potential for differentiating between benign and malignant tumors. To implement these advances in the clinic, the total frequency-dependent attenuation along the propagation path must be determined on a patient specific basis. Previously, an algorithm was developed to estimate this attenuation using echoes from multiple sources. In this study, the developed algorithm was extended to echoes from a single source by filtering the echoed signal into multiple frequency bands. This step was needed because it would be challenging to scan exactly the same tissue region using multiple sources in the clinic. Computer simulations and phantom experiments were conducted to verify the attenuation could be determined by filtering the echoes from a single source. The simulations utilized a spherically focused single-element source (5 cm focal length, f/4, 14 MHz center frequency, 50% bandwidth) exposing a homogeneous tissue region (Gaussian scattering structures with effective radii of 5 to 55 mum at a density of 250/mm(3), attenuation of 0.1 to 0.9 dB/cm.MHz). The phantom experiments utilized a spherically focused single-element source (5.08 cm focal length, f/4, 7.5 MHz center frequency) exposing a 0.5 dB/cm.MHz homogeneous glass bead phantom. The computer simulations and phantom experiment confirmed that the total attenuation along the propagation path can be determined by appropriately applying multiple filters to the backscattered echoes from a single source.