Abstract: A composition, device and method useful as an enzymatic urea assay based on the use of urea amidolyase comprising urea amidolyase, pyruvate kinase, pyruvate oxidase, mono and divalent cations, phosphoenolpyruvate, thyamine pyrophosphate, ATP, bicarbonate, phosphate, a color indicator system and optionally inorganic phosphate, a buffer having a pH range of from about 6.5 to 9.5, and sodium or potassium ferrocyanide. After contacting the test solution or body fluid sample with the assay composition, reading can be accomplished visually or instrumentally.