The aims of the present study were to examine whether collagen exists in egg, particularly in egg yolk; to establish a Fourier transform-near infrared (FT-NIR) measurement method for collagen in egg and to assess the possibility of increasing the collagen content by feeding hens a diet containing wood charcoal powder containing wood vinegar liquid (WCV). The collagen in eggs from 67-week-old hens fed on the dietary 0 and 9.9 g/kg WCV diets was investigated using a combination of histochemical, matrix-assisted laser desorption ionisation with time-of-flight mass spectrometry (MALDI-TOF MS), Fourier transform infrared (FT-IR) and FT-NIR approaches. All approaches used to identify collagen in egg yolk yielded positive results. The collagen in egg yolk measured using colorimetry, collagen in egg yolk, egg white and eggshell membrane using FT-NIR and collagen in egg yolk determined by treating the egg yolk with collagenase were abundant after feeding a dietary WCV (p<0.05). These results suggest that egg yolk contains collagen, that the collagen in egg can be measured using FT-NIR, and that the collagen content of egg yolk can be increased by feeding dietary WCV diets.