Objective To evaluate the effect of layering strategy and substrate color on the masking ability of resin composites. Materials and Methods A1-shaded specimens from Charisma Diamond and Filtek Z350XT were produced using different layering strategies. Color measurements were made by a reflectance spectrophotometer over A2, C2, A3.5, C3, C4 substrates. Color differences were calculated and interpreted by the 50%:50% perceptibility and acceptability visual thresholds. Data was analyzed by Kruskal-Wallis and Dunn post hoc test. Chi-square test was used to determine the association between masking ability, and independent variables. Results Color differences were significantly lower on A2 and C2 in comparison with C4 for the majority of the layering strategies. Acceptable matches were observed on most of the combinations over A2. Moderately unacceptable mismatches were observed in most of the combinations over C2 and A3.5. Clearly unacceptable mismatches were observed on the C3 and C4. The Delta E-00 color shifts were predominantly influenced by Delta L-00 for all layering strategies and substrate colors. Conclusion Masking ability was affected by the layering strategy and substrate color. Acceptable masking was associated with A2 and C2, and with layering strategy composed of 0.5 mm enamel opacity and 1.0 mm dentin opacity thicknesses, using the Filtek Z350XT. Clinical Significance Resin composites-shade A1-applied by different layering strategies with a final thickness of 1.5 mm were able to mask mild and moderately discolored substrates. Severely discolored substrates were not masked effectively.