Opaque white ink is highly important when printing on transparent substrates. The purpose of the white ink is to completely or partially cover the content of the packaging as well as to ensure the printing of other colors over it according to the appropriate printing specifications. The main goal of this study was to research the impact of anilox roller volume of opaque white on its opacity, and, consequently, on opacity, as well as the CIELAB values of colors that are printed over it. The research was conducted in three printing stages, during which the volume of the anilox roller was increased in linear steps of 3 cm3/m2 while other parameters remained constant. The analysis of white ink covering properties was carried out on microscopic print images using ImageJ software. The results showed that a linear increase in the anilox roller volume resulted in an exponential change in white ink opacity as well as the opacity of color samples. It was also determined that a volume increase of 6 cm3/m2 was required to visually notice color difference. Namely, medium lightness colors more strongly reacted to changes in white ink opacity compared to light and dark colors.