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The most significant uncertainty identified by the EU experts was the concern that TiO2 particles may have genotoxic effects. Genotoxicity refers to the ability of a chemical to directly damage genetic material within a cell (DNA), which may lead to cancer in certain situations. Although the experts did not conclude that TiO2 particles in E171 are genotoxic, they could not rule out the concern that they might be.

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The production process of lithopone involves a sophisticated synthesis of zinc sulfide and barium sulfate, resulting in a pigment that boasts high whiteness, opacity, and heat stability. The ratio of ZnS to BaSO4 can be adjusted to cater to specific applications, with some formulations containing up to 70% zinc sulfide and 30% barium sulfate. This flexibility makes lithopone an adaptable solution for manufacturers seeking optimal performance in their products.

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Mexican researchers sought to evaluate the effects of E171 across a span of conditions in mice, including its influence on behavior, along with the effects on the colon and liver. The research, published in 2020 in the journal Food and Chemical Toxicology, showed that E171 promoted anxiety and induced adenomas, or noncancerous tumors, in the colon. They also found that E171 heightened goblet cells hypertrophy and hyperplasia, which is typically seen in asthma patients and triggered by smoking or external pollutants and toxins. They also noted mucins overexpression in the mice, which can be linked to cancer cell formation. 

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