Calcium carbonate is a naturally occurring mineral that is found in rocks and minerals such as calcite, aragonite, and limestone. It is commonly used in industries such as paint, paper, plastics, and rubber as a filler and pigment. The manufacturing process of calcium carbonate involves extracting the mineral from quarries or mines, crushing and grinding it into a fine powder, and then purifying it through various chemical processes.

Lithopone, C.I. Pigment White 5, is a mixture of inorganic compounds, widely used as a white pigment powder. It is composed of a mixture of barium sulfate and zinc sulfide. These insoluble compounds blend well with organic compounds and confer opacity. It was made popular by the cheap production costs, greater coverage. Related white pigments include titanium dioxide, zinc oxide (zinc white), zinc sulfide, and white lead.^[1]

This constant high rate of ROS production leads rapidly to extreme macromolecular oxidation, here it is observed in the AOPP and MDA detected after 3 h in samples treated with bare P25TiO₂NPs (Fig. 6, Fig. 7). Macromolecular oxidation includes, among others, both protein and lipid oxidation. The ROS causes protein oxidation by direct reaction or indirect reactions with secondary by-products of oxidative stress. Protein fragmentation or cross-linkages could be produced after the oxidation of amino acid side chains and protein backbones. These and later dityrosine-containing protein products formed during excessive production of oxidants are known as advanced oxidation protein products (AOPP). They absorb at 340 nm and are used to estimate the damage to structural cell amino acids. Lipid oxidation is detected by the conjugation of oxidized polyunsaturated lipids with thiobarbituric acid, forming a molecule that absorbs light at 532 nm. Polyunsaturated lipids are oxidized as a result of a free-radical-mediated chain of reactions. The most exposed targets are usually membrane lipids. The macromolecular damage could represent a deadly danger if it is too extensive, and this might be the case. Moreover, it could be observed that cellular damage continues further and becomes irrevocable after 6 h and MDA could not be detected. This may be due to the fact that the lipids were completely degraded and cells were no longer viable. Lipids from the cell membrane are the most prone to oxidation. In fact, lipid peroxidation biomarkers are used to screen the oxidative body balance [51]. At the same time, AOPP values are up to 30 times higher for bare nanoparticles in comparison to the functionalized ones.