With the ever-increasing use of pharmaceuticals and the subsequent release into the environment comes the need for reliable detection methods. While the emphasis continues to be on chromatographic methods, every time, there are more and more biologically based assays being developed for the detection of pharmaceuticals in the environment. This chapter affords an overview of the currently available bioassays, biochemical assays, and biosensors focusing on the most commonly used pharmaceuticals-antibiotics, hormones, and analgesics. Bioassays rely on biological indicators, whether it be cells, tissues, or whole organisms, to evaluate an analyte according to its biological activity, whereas biochemical assays and biosensors incorporate biorecognition elements such as enzymes, proteins, or antibodies but the signal is enhanced by physical or chemical means. There has been more emphasis given on biosensors, especially immunologic sensors, for as the reader will appreciate, they show greater promise as alternatives to chromatography as they are fast, robust, specific, and quantifiable and may be performed on-site.