Defining virtual elimination has created considerable debate. A traditional approach has been to use chemically defined detection limits or levels of quantification that are determined using the best currently available methodologies. Ever increasing improvements in analytical techniques could lead to corresponding pressure to reduce the targets for virtual elimination. The current Toxic Substances Management Policy in Canada recognizes this and clearly states that it is not the intent of virtual elimination to have a moving target or to chase down the last molecule of the chemical of concern. Although it may be possible to reduce a chemical to less than some extremely sensitive detection limit, the chemical may or may not exert biological effects at that level. The chemically defined detection limits may be much lower than background levels in the environment, making it an unrealistic target. Conversely biological responses may result from trace levels of a compound that are not detectable in effluents or selected compartments of the environment (i.e., water) using current chemical techniques. Alternatively, an effect-based approach can establish biologically meaningful endpoints to defining virtual elimination. Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are used in this study as an example to evaluate the advantages and limitations of several possible approaches of using biological endpoints to determine the presence of these compounds in the environment and ultimately define virtual elimination. A review of the biological responses to PCDD/PCDFs is included to demonstrate the importance of selecting appropriate biological endpoints. Mixed function oxygenase (MFO) induction, although not recommended at this point, is used as an example of a possible sensitive endpoint that could potentially be used to detect exposure of biota to these chemicals. Three different approaches are explored: (1) measuring MFO induction in a sentinel species in the environment; (2) testing environmental extracts for MFO induction in cell lines; and (3) using biological endpoints (MFO induction) to define chemical targets for virtual elimination. While the use of biological end-points is the most desirable approach to defining virtual elimination, there are significant knowledge gaps which limit our selection and application of this approach.