All color-difference formulas are developed to evaluate color differences for pairs of stimuli with hair-line separation. In printing applications, however, color differences are frequently judged between a pair of samples with no-separation because they are printed adjacent on the same piece of paper. A new formula, dENS has been developed for pairs of stimuli with no-separation (NS). An experiment was conducted to investigate the effect of different color-difference magnitudes using sample pairs with NS. 1,012 printed pairs with NS were prepared around 11 CIE recommended color centers. The pairs, representing four color-difference magnitudes of 1, 2, 4 and 8 CIELAB units were visually evaluated by a panel of 19 observers using the gray-scale method. Comparison of the present data based on pairs with NS, and previously generated data using pairs with hair-line separation, showed a clear separation effect. A new color-difference equation for the NS viewing condition (dENS) is proposed by modifying the CIEDE2000 formula. The separation effect can be well described by the new formula. For a sample pair with NS, when the CIEDE2000 color difference is less than 9.1, a larger color difference leads to a larger lightness difference, and thus the total color difference increases. When the CIEDE2000 color difference is greater than 9.1, the effect is opposite, i.e. the lightness difference decreases, and thus the total color difference also decreases. The new formula is recommended for future research to evaluate its performance in appropriate applications.