Introduction The influence of dehydration on tooth shade constitutes a significant aspect, making it a pivotal area of study, particularly within the field of restorative dentistry. This prospective study was done to analyze, using a spectrophotometer, how dehydration affects the shade of the teeth. Materials and methods Twenty-five participants were recruited in this prospective study that took place in the Dental Care Center of Saint Joseph University of Beirut (Lebanon) between October 2022 and January 2023. The inclusion criteria for this study primarily comprised four intact maxillary incisors. On the other hand, damage, cavities, restorations, and staining served as exclusion criteria. The VITA Easyshade Advance (Vident,Brea,CA) spectrophotometer was used to record the measurements. The measures of L (which represents darkness to lightness), a (which represents greenness to redness), and b (which represents blueness to yellowness)were registered at the center of each tooth's labial surface at baseline and then at 10-minute intervals for 30 minutes while the teeth were dehydrating due to the placement of a rubber dam. The color difference (ΔE) was later calculated using L*a*b* measures, with the perceptibility threshold set at ΔE00=0.8. Statistical analyses were performed using the Friedman test, the Bonferroni post hoc test, and the Wilcoxon signed-rank test. Results The different color changes represented byΔE(ΔE1, ΔE2, and ΔE3) were found to be significantly higher than the perceptibility threshold of 0.8 (p<0.001). A statistically significant difference was found between ΔE1 and ΔE3 (p<0.05). Moreover, a statistically significant difference was found between L0, L10, L20, and L30 (p<0.001). Statistically significant differences (p<0.05) exist between the means of L0 and L20, the means of L0 and L30, the means of L10 and L20, and the means of L10 and L30. Conclusion This study showed that dehydration affected the shade of the teeth: Lightness increased, and therefore, the teeth appeared whiter. Dentists should consider the hydration status of the teeth when evaluating color for treatments, as dehydration can significantly affect shade matching: the more the dehydration time elapses, the more the color difference compared to the baseline increases.