Although the estrogenic hormones 17 beta-estradiol and 17 alpha-ethinyl estradiol can be quantified in polluted waters by gas chromatography/mass spectrometry (GC/MS) and gas chromatography/tandem mass spectrometry (GC/MS/MS), the compounds often are present at concentrations below detection limits. Enzyme-linked immunosorbent assays (ELISAs) provide a sensitive and robust means of quantifying estrogenic hormones in wastewater effluents and surface waters. Results from ELISA analysis of estrogenic hormones in secondary wastewater effluent indicate concentrations comparable to those that cause vitellogenesis in fish. Confirmatory analyses by GC/MS/MS are consistent with ELISA results. Effluent filtration, using sand filtration or microfiltration, removes approx. 70% of the hormones from secondary effluent, while advanced treatment, using reverse osmosis, removes more than 95% of hormones. The detection limits for estrogenic hormones are approx. 0.1 ng/L in wastewater effluent and 0.05 ng/L in surface water. The ELISA technique provides a relatively simple and practical method of assessing the fate of estrogenic hormones in engineered and natural systems.