Glyphosate (N-(phosphonomethyl)glycine) is the most widely used herbicide on earth. A simple assay to quantify glyphosate concentrations in environmental samples was developed as part of an interdisciplinary effort linking introductory laboratory courses in chemistry, biology, and microbiology. In this 3 h laboratory experiment, students used UV-vis spectroscopy to quantify glyphosate in prepared unknowns and supernatants from glyphosate-treated soil samples. Regression analysis indicated that the assay is linear up to 20.0 ppm, making it particularly useful for detection of low levels of glyphosate in environmental samples. The assay can be used to quantify the activity of glyphosate-degrading soil microorganisms by comparing glyphosate levels between autoclaved and nonautoclaved soil slurries.