The activity and kinetics of N-acetyltransferase (NAT) in the quail duodenum were studied by radioenzymatic assay. NAT activity was highest when incubated under 37 degrees C, at pH 5.8 for 15 sec. Of the four substrates tested (tryptamine, 5,6-dihydroxytryptamine, serotonin, and N-acetylserotonin at concentrations of 0.08-4 mM), only tryptamine showed the substrate saturation phenomenon when incubated with the duodenal enzyme and acetyl-14C-coenzyme A. The saturation concentration of tryptamine was about 4 mM. Using the double reciprocal plot and regression equation, the Michaelis constant (Km) and maximal rate (Vmax) of NAT activity were found to be 0.204 mM and 0.917 nmol.mg protein-1.min-1, respectively. Diurnal study demonstrated higher NAT activity at middark (3.7 nmol.mg protein-1.min-1) and lower activity at midlight (2.4 nmol.mg protein-1.min-1), suggesting a circadian rhythm of the enzyme activities in the quail duodenum.