The study of the role of carotenoids on the physiology and evolutionary ecology of birds demands methods for their quantification in the bloodstream. We compared color-chart scores of plasma hue with the actual concentration of plasma carotenoids obtained by spectrophotometry in 356 wild birds from 26 species. Repeatability of chart scores between three independent observers was high. However, color scores did not correlate with the spectrophotometric results in interspecific analyses. Within species (n = 3), one showed no relationship and two showed weak but significant positive correlations. Hemoglobin, and probably other substances, may mask the color of carotenoids, making the accurate use of color charts difficult. Spectrophotometry should be the method of choice as it permits precise quantifications of total plasma carotenoids and objective comparisons among studies.