The production of ROS was studied on white blood cells as a model to screen the effect on eukaryotic cells after being exposed to samples and solar simulated irradiation (according to the level of penetration under the skin). For that purpose, the leukocytes were separated from anticoagulated fresh blood using the Ficoll-Hypaque reactive in a well-known technique [33]. Then, 50 μL of suspensions of P25TiO₂NPs (0.2 mg/mL and 0.02 mg/mL), vitaminB2@P25TiO₂NPs (0.2 mg/mL and 0.02 mg/mL) and vitamin B2 (0.2 mg/mL and 0.02 mg/mL) were prepared and mixed with 50 μL of white blood cells suspension. A solution of 3% H₂O₂ was used as positive control and PBS as negative control. Then, the samples were irradiated using the LED panel for 3 and 6 h to simulate the light penetration into the skin. Also, a set of samples was kept in the dark as control. Finally, the ROS were detected through the colorimetric assay employing the nitroblue tetrazolium salt (NBT salt) and the absorbance at 650 nm was measured. The experiment was reproduced twice; the standard deviation was calculated and p-value < 0.05 were considered significant.

Fig. 6. AOPP measured on samples of MSSA with: A) 0.2 mg/mL P25TiO₂NPs; B) 0.02 mg/mL P25TiO₂NPs; C) 0.2 mg/mL VitaminB2@P25TiO₂NPs; D) 0.02 mg/mL VitaminB2@P25TiO₂NPs after 3 h of irradiation (red) and 6 h (blue). SD <1 (error bars too small to be seen) and p < 0.05 between C-D and A-B.

Following six months of phasing out the additive, titanium dioxide will be completely banned in the European Union starting August 7. France had previously banned the use of titanium dioxide in food starting in January 2020.

Lithopone