Infrared analysis showed that the characteristics bands for the bare nanoparticles are still exhibited in the vitamins@P25TiO₂NPs spectra, such as a wide peak in 450–1028 cm⁻¹ related to the stretching vibration of Ti-O-Ti and other peaks in 1630 cm⁻¹ and 3400 cm⁻¹, which represent the surface OH groups stretching. The IR spectrum of vitaminB2@P25TiO₂NPs showed signs of binding between compounds. The OH bending peak (1634 cm⁻¹) corresponding to bare nanoparticles disappeared, and the NH₂ bending band characteristic of vitamin B2 appeared (1650 cm⁻¹). The IR spectrum of vitaminC@P25TiO₂NPs also showed signs of successful functionalization. Bands at 1075 cm⁻¹; 1120 cm⁻¹; 1141 cm⁻¹ were observed, which are originated by CO-C vibrations present in the vitamin C. The intense band at 1672 cm⁻¹ is attributed to the C = O stretching in the lactone ring while the peak at 1026 cm⁻¹ is ascribed to the stretching vibration Ti-O-C. Wide bands at 3880–3600 cm⁻¹ are related to stretching vibration OH groups, but those disappear in the modified nanoparticles spectrum. These observations confirm the interactions between the P25TiO₂NPs and the vitamins [35].

Nano-sized TiO₂ generally shows low or no acute toxicity in both invertebrates and vertebrates. However, exposure of Daphnia magna to 20 ppm TiO₂ for 8 consecutive days was found to cause 40 % mortality. Zhu et al. showed minimal toxicity to D. magna after 48 h exposure, while upon chronic exposure for 21 days, D. magna suffered severe growth retardation and mortality. A significant amount of nano-sized TiO₂ was found also accumulated in the body of the animals. Similar findings with coated nano-sized TiO₂ (T-Lite™ SF, T-Lite™ SF-S and T-Lite™ MAX; BASF SE) were reported by Wiench et al. Biochemical measurements showed that exposure to TiO₂ NPs induces significant concentration-dependent antioxidant enzyme activities in D. magna. Lee et al. showed that 7 and 20 nm-sized TiO₂ induced no genotoxic effect in D. magna and in the larva of the aquatic midge Chironomus riparius.