Photopic electroretinograms (ERGs) elicited by light flashes were recorded for three normal human subjects who were exposed, throughout, to natural ambient light cycles over 24-h day–night periods. ERGs were recorded either with the adaptation state of the eyes maintained at the level set by the natural ambient lighting, or after 10 min dark adaptation. The amplitudes and implicit times of both the a- and b-wave components of the ERG were examined and of these, only the b-wave implicit time exhibited significant diurnal variation, such that the nighttime values were 20–40% greater than those recorded during daytime. Such diurnal variations were observed under both recording conditions and cannot, therefore, be attributed to diurnal changes in the adaptation state of the cone photoreceptors. ERGs were recorded at midday and midnight during 24-h exposure to the natural light cycle, but during the recording period, the short-term adaptation state of the eye was controlled by exposure to rod saturating background field, so that visual sensitivity was the same at both recording times. The b-wave implicit times recorded at midnight were, nonetheless, greater than those recorded at midday. This difference is not, therefore, determined by the short-term state of retinal adaptation, but reflects long-term light history. Measurements performed under 24-h continuous light exposure showed no variation in the b-wave implicit time, whereas some measurements made during extended dark adaptation provided limited evidence for implicit time changes. By controlling the wavelengths to which the eye was exposed during the daylight phase of the diurnal cycle, it was shown that the shifts in b-wave implicit time associated with the change from dark to light are triggered by the rod system, although they are most clearly observed in the cone-dominant responses to long-wavelength light. The results demonstrate a diurnal variation in the temporal responses of the post-photoreceptoral cone pathways of the human retina, which is triggered by activation of the rod photoreceptors.