Bisphenol A (BPA; C15H16O2, purity >99.5%) was purchased from the Chemical Agent Co. Protein Tyrosine Kinase inhibitor (Tianjin, China). A BPA stock solution was find more prepared in an aqueous solution that contained ethanol as a co-solvent. The final concentration of ethanol was less than 1%, and our previous

evaluations had demonstrated no adverse biological effect at this concentration. The stock solution was then diluted to specific test concentrations with the dilution solvent. Adsorption experiments of BPA on TiO2-NPs in vitro: analysis of BPA concentration before and after mixture To analyze if there were changes of BPA concentration after mixing with TiO2-NPs, different concentrations of BPA solutions (5 and 10 mg/L) and combined solutions (BPA 5 mg/L + TiO2 10 mg/L, BPA 5 mg/L + TiO2 50 mg/L, BPA 10 mg/L + TiO2 10 mg/L, and BPA 10 mg/L + TiO2 50 mg/L) were prepared. Then the combined solutions were added to 50-mL plastic centrifuge tubes (in a preliminary experiment, the plastic centrifuge tubes were proven to have no effect on the BPA concentration). Subsequently, the tubes were centrifuged for 10 min Ilomastat datasheet at 14,000 rpm using a high-speed centrifuge (HIMAC CR 22G II, Hitachi, Ltd., Chiyoda-ku, Japan), and the supernatant was collected and centrifuged again at 14,000 rpm. Before injection, the BPA solutions and the combined solutions were passed through 0.22-μm filter membranes, respectively. The samples were analyzed by high-performance

liquid chromatography (Waters 2695 HPLC, Waters Corp., Milford, MA, USA) with a photodiode array detector (Waters 2998 PDA) for BPA levels. The samples were separated on a CAPCELL PAK C18 column (4.6 × 150 mm, 5 μm, Waters Corp.) using a mobile phase of 60% acetonitrile and 40% water at a flow rate of 1.0 mL/min. The Calpain injection volume was 10 μL, and the column temperature was 20°C. The pH values for all of the samples before and after the adsorption experiments were similar and were approximately neutral. Adsorption experiments of BPA on TiO2-NPs in vitro: adsorption kinetics of BPA on TiO2-NPs A solution of BPA 5 mg/L + TiO2

10 mg/L was used to measure adsorption kinetics. The combined treatment solution was shaken at 250 rpm in a reciprocating shaker at 20°C ± 1°C. The kinetic data were collected with an initial BPA concentration of 5 mg/L. Subsequently, 10 mL of the treatment solution was collected and centrifuged twice at 14,000 rpm at 0, 5, 10, 30, 60, 90, 120, and 180 min, and the BPA concentrations were analyzed. Zebrafish maintenance and embryo collection Juvenile zebrafish were purchased from a local aquarium in Tianjin, China. The fish were kept at aquarium conditions of 26°C ± 1°C at a density of ≤1/L using charcoal-filtered tap water. The pH of the maintenance water was within the range of 6.8 to 8.4. Oxygen saturation in the maintenance aquaria was kept above 80%. The fish were maintained with a 14:10-h light/dark cycle and fed dry flake food and frozen midge larvae twice or thrice per day.