Statement of problem: Environmental conditions play a major role for accurate determination of tooth color. However, often underestimated is the impact like intensity of illumination, individual adaptation time as well as the influence of environmental conditions on color determination? Purpose: The aim of this research is to analyze the influence of background color on the visual determination of tooth color. Material and methods: Dental students (N=37) at the Universities in Dresden, Leipzig, Berlin, Olomuc and Greifswald participated in this research. A standardized lecture, followed by the web-based Toothguide-Trainer tool (www.toothguide.de) prepares the students for templates matching with the computer aided "Tooth guide-Training-Box" (TTB, VITA Zahnfabrik).After color matching in neutral grey surrounding (reference), the participants repeated the matching on modified TTBs (red, yellow and blue). Color distance to the selected templates (∆E) was calculated confounded by gender and age. The mean and standard deviation (SD), median and the 95 % confidence interval (95% CI) were calculated using Mann-WhitneyU test (α=.05). Results: Compared with the reference (neutral grey) a color distance (mean ΔE) of 2.45 and standard deviation (SD) ± 2.6, ΔE for red background was 3.35 (± 2.8), blue had a ∆E value of 2.98 (± 2.6). U test showed highest significant differences (P < 001) to the reference. Yellow background showed a mean of ΔE 2.94 (± 2.7) with P <.01. 95% CI for reference was 2.16-2.49, red 3.07-3.44, blue 2.71-3.05 and yellow 2.67-3.02.Odds ratio of misassessment (∆E > 3) for red environment was 2.14 (P < 001), for blue background with 1.73 (P < 001) and yellow with 1.53 (P =.001) significant higher compared with reference (neutral grey).For all background colors ("yellow", "blue", "red"), men needed a significantly longer evaluation time (P < 05, respectively) in color differentiation than for the grey background (reference). Conclusion: The environmental conditions play a major role and influence the visual determination of tooth color significantly. Standardized conditions are required to get exact results in the tooth color differentiation. Clinical implications: Color determination requires "dazzle-free" standard environment conditions (neutral grey environment, day light illumination) to improve the visual color differentiations procedures. Eliminable factors, like colorful outerwear, make-up, colorful walls and ceilings as well as colorful dental instruments have to be avoided.