Heparinized whole-blood samples from 22 adult African lions (Panthera leo) fed diets considered nutritionally adequate in 10 American Zoo and Aquarium Association member zoos in North America were provided for this study. Blood thiamine values were estimated using a standard microbiological assay method. The mean ± standard deviation for blood thiamine values was 249.3 ± 43.5 nmol/L with a range in values from 160 to 350 nmol/L after exclusion of one outlier. There were no differences (P > 0.05) in the mean blood thiamine values of male and female lions, or of lions that were over and under 10 yr of age. This range (160 to 350 nmol/L) is proposed as a reasonable estimate of the expected range in blood thiamine values for captive adult African lions as currently fed in North American zoos.