Polychromatic ultraviolet irradiation, such as from medium pressure (MP) Hg lamps may enhance the UV degradation of environmental pollutants as compared to low pressure (LP) Hg UV sources emitting monochromatic irradiation. Typically, studies involving destruction of environmental pollutants such as endocrine disrupting compounds (EDCs) are based on measurement of the parent compound decay using analytical chemistry, but such information is insufficient to determine an effective treatment endpoint because the identity and biological activity of many transformation products remain unknown. Bioanalytical methods to assess residual biological activity of a treated water offers one means to compare removal efficiency of EDC activity between MP- and LP-UV lamps under photolysis and UV/H2O2 oxidation. In this study, changes in estrogenic activity of bisphenol-A (BPA) as a function of UV treatment were evaluated using both an in vitro yeast estrogen screen and in vivo vitellogenin assay with Japanese medaka (Oryzias latipes) fish. Decay of BPA parent compound and formation of degradation products were followed using HPLC analysis. Results demonstrated that MP-UV direct photolysis more effectively removed BPA and associated estrogenic activity compared to LP-UV lamps. UV in combination with H2O2 significantly removed estrogenic activity in vitro and in vivo compared to direct photolysis; however, no significant difference in removal rates was found between the two lamps under UV/H2O2 oxidation. Furthermore, the UV/H2O2 process was effective for reducing embryo toxicity of BPA, but resulted in the production of acidic intermediates, causing acute toxicity and delayed hatching in some medaka embryos.