Enantioselective analysis of some pharmaceuticals during wastewater treatment has the potential to reveal significant insights regarding the effectiveness of biotransformation processes. Furthermore, enantioselective analysis of chiral pharmaceuticals in the aquatic environment may provide a useful historical record revealing the dominant source of (treated or untreated) wastewater contamination. This review of the recent scientific literature has identified only a handful of studies that have directly investigated these promising applications. However, a range of enantioselective analytical techniques are likely to be adaptable from those which have been developed within the pharmaceutical industry. These include direct enantioseparations of enantiomers on chiral stationary phases as well as indirect separations by achiral stationary phases after chiral derivatization to form pairs of physically distinguishable diastereomers. Further investigations of the patterns of enantiomeric fractionation of pharmaceuticals in wastewater and environmental samples will provide an increasingly solid understanding of the relationship between biotransformation processes and the often overlooked parameter of enantiomeric fraction.