Abstract Background The analytical performances of the Ammonia Ultra assay for in vitro quantitative enzymatic determination of Ammonia in human plasma have been evaluated on Beckman AU/DxC AU Systems. Ammonia, derived from the amino acids catabolism and from the action of intestinal bacteria on dietary proteins, is converted to non-toxic urea by the liver. An excess of ammonia can have toxic effects on the Central Nervous System with possible neurological disturbances. Elevated ammonia levels can also be observed in severe liver failure as may occur in Reye’s syndrome, viral hepatitis or cirrhosis. Methods Ammonia, in the presence of glutamate dehydrogenase (GLDH), combines with alpha-ketoglutarate and NADH to yield glutamate and NAD+. The decrease in 340 nm absorbance (NADH → NAD+) is proportional to the ammonia level in the examined plasma. The reagent contains excess of lactate dehydrogenase (LDH) to rapidly reduce endogenous pyruvate and avoid its interference with the assay system. Performances evaluation have been done following current CLSI guidelines protocols. Results 3 calibrator lots and 2 reagent lots have been used for testing. For each test and among different combinations of reagents and calibrators, worst case has been reported in the table. Conclusions Analytical performances of Ammonia Ultra assay on Beckman AU/DxC AU Systems meet the requirements for its use as quantitative determination of Ammonia in human plasma and make this assay very suitable for the routine measurement of this analyte.