Hops have been used for centuries to impart aroma and bitterness to beer. The cones of the female hop plant contain both essential oils, which include many of the fragrant components of hops, and a collection of compounds known as α- and β-acids that are the precursors to bittering agents. In order for brewers to predict the ultimate bitterness of a brewed product, it is important to have an accurate assessment of the α- and β-acid content of the hops. A high-performance liquid chromatography method is described here for the quantitation of the predominant α-acids (humulone, cohumulone, and adhumulone) and β-acids (lupulone, colupulone, and adlupulone) present in hops. Additional laboratory projects are suggested that allow students to explore various aspects of method assessment and validation.