Fiber color in cotton is a genetically inherited trait resulting from the presence of pigments intermingled with cellulose. Electronic shade-matching instruments can be used to determine objectively the color of cotton fiber. The objectives of this study were to investigate brown to white fiber color segregation in the F2 population of ‘Carmen’ (white fiber) x ‘Devetüyü-176’ (brown fiber) using an electronic shade-matching instrument (chromameter) and compare those results to visual color analysis. The results demonstrated that transgressive segregations were identified for all fiber color parameters in the F2 population. The fiber color parameters (L, ∆L, a, b, ∆a, ∆b, and ∆E) and the visual analysis were considered separately. Each parameter appeared to be controlled by a different single gene displaying complete or partial dominance of the brown color over white except L and ∆L. In summary, the results obtained from the use of the chromameter will aid in the reproducible and quantifiable determination of cotton fiber color and reduce observation errors inherent in visual analysis. This would be most useful in colored cotton breeding programs and in selection and in the monitoring of color drift.