Steroidal estrogens in the environment exert toxicological effects at very low concentrations. Furthermore, dissolved and colloidal fractions of soil and manure play an important role in the environmental fate and transport of steroidal estrogens. One objective of this study was to quantify the association of the natural estrogen, 17β-estradiol (E2), with the dissolved fraction and colloidal fraction isolated from liquid swine manure (LSM), soil, and soil+LSM mixtures. The second objective of this study was to evaluate whether the E2 associated with the dissolved fraction/colloidal fraction, dissolved fraction and colloidal fraction of the various media could induce an estrogenic response. Estrogenicity was assessed using an E2 receptor (ER) competitor assay, which provided E2 equivalent concentration (EEQ) of dissolved fraction/colloidal fraction, dissolved fraction and colloidal fraction solutions created from the Soil, Soil+LSM and LSM.