AbstractThe purpose of this study is to verify whether the spectrophotometer makes similar color readings when subjected to the use of biosafety barriers. Eighty specimens were made in four colors of the Llis composite resin (EA1, DA1, EA3, DA3) and four of the Filtek Z350 XT (A1E, A1D, A3E e A3D) (n = 10). The specimens were subjected to color analysis with a VITA Easyshade device according to the groups regarding the physical barrier: the protector supplied with the VITA device, polyvinyl chloride plastic film, high‐density polyethylene bag, and without protection as a control. The color difference (ΔE00) between control and groups was evaluated and compared with the perceptibility and acceptability thresholds. In the Filtek Z350 XT, there was a significant ΔE00 in color A3E, in the polyethylene bag (P < .001), and in color A1D, in the supplied protector (P = .01) in relation to the other groups. For Llis, in color EA1 for all protective barriers (P = .014), and in color DA1 only for polyethylene bag in relation to the other barriers (P = .02). ΔE00 was higher than the perceptible threshold for all groups, colors, and brands. The results below the threshold of acceptability in relation to the ΔE00 did not demonstrate consistency between barriers, colors, and resins. It was not possible to identify a statistically significant pattern of ΔE00 between the barriers. The ΔE00 were above the perceptibility threshold, and few were below the acceptability threshold. It was not possible to suggest that one biosafety barrier was superior to another or could be indicated.