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EC number: 236-675-5
CAS number: 13463-67-7
bioaccessibility of titanium dioxide was determined in vitro by
simulating dissolution in the cell culture medium (MDEM), required for
culturing lung epithelial cell lines (FE1 cells derived from
MutaTMMouse) used for in vitro testing of chemical induced mutagenicity,
with a loading of and
10 mg/L (nano-TiO2) and 100 mg/L (nano- or bulk-TiO2). Dissolved
concentrations were operationally defined as the dissolved titanium
fraction from the top part of supernatant after centrifugation (20000xg,
3 x 30 min), see table 2.
After 24 hours, dissolution of nano-TiO2 was highest in cell culture
medium (MDEM) at an initial concentration of 10 mg/L.
2: Released amounts of TiO2 ENMs into water and DMEM (metal oxide
initial concentration: 10 mg/L and 100 mg/L presented as mean (standard
deviation) of triplicates. The results are expressed as mg/L and as
percent metal released.
LOD: below limit of detection
Table 3 shows the influence of
particle size on percent solubility of nano and bulk TiO2 after 24-h
incubation in deionized water and cell culture medium at an initial
metal oxide concentration of 100 mg/L.
Influence of particle size (nano versus bulk) on % solubility of TiO2
after 24-h incubation in water and DMEM (Initial TiO2 concentration of
0.008 (0.001) %
0.045 (0.012) %
0.0002 (0.00001) %
0.0005 (0.0003) %
presented as mean (standard deviation) of triplicates.
Method validation summary
detection (LOD) for titanium: 0.00018 mg/L (in water); 0.00017 mg/L (in
range for TM 24.4 and TMDA 64.3 trace element reference materials was
80–105 % for titanium. Samples were matrix blank corrected.
Mean recovery of
fortified samples: 88.6–109.7 %
Mean recovery of
fortified samples: 93.4–104.2
The objective of
this study was to investigate the dissolution of three metal
oxide-engineered nanomaterials (ENMS) (CuO, NiO, and TiO2) in water
versus cell culture medium.
The test was performed on the basis of OECD
Series on Testing and Assessment No. 29 (2001; ENV/JM/ MONO(2001)9),the
Standard Operating Procedure for Bioelution Testing of Metals, Inorganic
Metal Compounds, and Complex Metal-Containing Materials: Simulated
Gastric Fluid (version 09.11.2018 submitted to ECVAM for acceptance) and
according to the bioaccessibility test protocol provided by the monitor.
The bioaccessibility protocol has been developed on the basis of
relevant published methods.
bioaccessibility protocol has been developed on the basis of relevant
The test medium
was selected to simulate the test conditions of an in vitro test which
is used for testing chemical-induced mutagenicity.
The test item was
pre-wetted in deionized water (1 mL) to form a wet paste and after 20 h
was dispersed in deionized water to yield the required concentration.
Stock dispersions of metal oxide ENMs (1000 mg/L metal oxide) were
prepared in deionized water and sonicated according to the material
specific delivered sonication energy (DSE). Freshly prepared stock ENM
dispersions (1000 mg/L and 100 mg/L) were diluted 10 times either in
deionized water or cell culture media (DMEM).
flasks per test item (TiO2 nano or bulk-form) and per test medium
(deionized water or DMEM) were prepared with loading of 10 mg/L or 100
test item concentration was specified by measuring the titanium
concentration of the test media (deionized water or cell culture medium
(DMEM)) under the applied test
conditions after the separation of undissolved test item by
centrifugation (20000x g, 3 x 30 min). However, it is
questionable if centrifugation at 20000xg for 3 x 30 min would be
sufficient for phase separation of the nano part.
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