Glare is an unwanted optical phenomenon which affects imaging systems with optics. This paper presents for the first time a set of hyperspectral image (HSI) acquisitions and measurements to verify how glare affects acquired HSI data in standard conditions. We acquired two ColorCheckers (CCs) in three different lighting conditions, with different backgrounds, different exposure times, and different orientations. The reflectance spectra obtained from the imaging system have been compared to pointwise reference measures obtained with contact spectrophotometers. To assess and identify the influence of glare, we present the Glare Effect (GE) index, which compares the contrast of the grayscale patches of the CC in the hyperspectral images with the contrast of the reference spectra of the same patches. We evaluate, in both spatial and spectral domains, the amount of glare affecting every hyperspectral image in each acquisition scenario, clearly evidencing an unwanted light contribution to the reflectance spectra of each point, which increases especially for darker pixels and pixels close to light sources or bright patches.