Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-03-04 to 2008-04-02
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented, scientifically sound GLP study performed according to OECD Guideline 471 and EU Method B.13/14.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Certificate provided by Rheinlandpfalz
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Zirconium dioxide
Cas Number:
1314-23-4
Molecular formula:
ZrO2
IUPAC Name:
Zirconium dioxide
Constituent 2
Reference substance name:
Zirconium dioxide
EC Number:
215-227-2
EC Name:
Zirconium dioxide
IUPAC Name:
215-227-2
Details on test material:
- Name of test material (as cited in study report): CC10 Zirconium Oxide
- Molecular formula (if other than submission substance): ZrO2 + HfO2
- Molecular weight (if other than submission substance): 123.22 g/mol
- Physical state: white powder
- Stability under test conditions: not stated
- Storage condition of test material: stored in a closed vessel at room temperature (20 +/- 5 degree C)

Method

Species / strain
Species / strain / cell type:
S. typhimurium, other: TA97a, TA98, TA100, TA102, TA1535
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
4998, 1499, 500, and 50 ug/plate- Experiment one
4998, 2499, and 1250 ug/plate- Experiment two
As the test item wasn't soluble in any suitable solvent, a stock suspension containing 50 g/L was prepared and diluted as necessary.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO; water
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-Nitro-1,2-phenylene diamine in DMSO (without at 80 ug for strains TA 97a, TA98 and TA102); Sodium azide in deionised water (without at 6 ug for strains TA100 and TA1535)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Benzo-a-pyrene; 2-Amino-anthracene in DMSO (with at 40 ug for stain TA98); 2-Aminoanthracene in DMSO (with at 3 ug for strains TA97a, TA100, TA102 and TA1535)
Details on test system and experimental conditions:
METHOD OF APPLICATION:

- in agar (plate incorporation)- Experiment one
Per strain and dose, four plates with and four plates without S9 mix were used. 10 mL of the test solution of the appropriate concentration were membrane filtrated (size of pores was 0.2 µm) into sterile vessels. Top agar basis was melted in a microwave oven, after melting, 10 mL of histidine-biotin-solution 0.5 mMol per 100 mL basis was added and the bottle was placed in the water bath at 45 degree C.
0.1 mL of the appropriate solution of the test item were given into a sterile tube. After mixing with 0.1 mL overnight culture of the respective strain and 0.5 mL phosphate buffer (only for treatments without S9) or 0.5 mL S9 mix, 2 mL Top-Agar were added. The mixture was gently vortexed, then poured on a minimal glucose plate and distributed evenly, using a Drigalski spatula. The plates were closed, covered with brown paper and left to harden for a few minutes, then inverted and placed in the dark incubator at 37 degree C.

- pre-incubation- Experiment two
Per strain and dose, four plates with and four plates without S9 mix were used. 10 mL of the test solution of the appropriate concentration were membrane filtrated into sterile vessels. Top agar basis was melted in a microwave oven, after melting, 10 mL of histidine-biotin-solution 0.5 mMol per 100 mL basis was added and the bottle was placed in the water bath at 45 degree C.
0.1 mL of the appropriate solution of the test item was given into a sterile tube. After mixing with 0.1 mL overnight culture of the respective strain, 0.5 mL phosphate buffer (only for treatments without S9) or 0.5 mL S9 mix were added. The mixture was incubated in an incubation chamber at 37 degree C for 20 minutes. During this time the vessels were aerated through careful shaking. Then 2 mL top agar was added. The mixture was vortexed gently, then poured on a minimal glucose plate and distributed evenly, using a Drigalski spatula. The plates were closed, covered with brown paper and left to harden for a few minutes, then inverted and placed in the dark incubator at 37 degree C.


DURATION
- Preincubation period: 20 minutes at 37 degree C
- Exposure duration: 48 hours at 37 degree C- Both experiments


NUMBER OF REPLICATIONS: 4


NUMBER OF CELLS EVALUATED: at least 10^9 cells/mL correlating to 100 colonies / plate




Evaluation criteria:
A test substance is considered to have mutagenic potential, if a significant, reproducible increase of revertant colonies per plate (increase factor >/= 2) in at least one strain can be observed. A concentration-related increase over the range tested can also be taken as a sign of mutagenic activity.
Statistics:
The colonies were counted visually, the numbers were recorded. A spreadsheet software (Microsoft Excel) was used to calculate mean values and standard deviations as well as the increase factor of revertant induction.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 97a
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
- The test item didn't show mutagenic effects in both experiments. The number of revertant colonies was not increased in comparison with the spontaneous revertants (solvent only).

- Cytotoxicity of the test item was not detected. The background lawn was visible and the number of revertants was not significantly decreased.

- No toxicity was observed


Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Mean Revertants First Experiment:

Strain     97a    98    100    102    1535  
 Induction    -S9  +S9  -S9  +S9  -S9  +S9  -S9  +S9  -S9  +S9
 H2O  Mean  139  109  13  11  152  185  212  192 15   16
   sd 58.2   11.6  1.7  3.8  25.1  31.1  22.0  58.2  3.6  5.7
 DMSO  Mean  136  155  8  10  206  178  204  221  18  15
   sd  16.8  49.5  4.7 3.4  30.2  34.9  12.4  73.3  2.4  5.4
 Pos Contr  Mean  1001  1001  1001  1001  1001  1001  1001  1001  1001  1001
   sd  0  0  0  0  0  0  0
  f(I)  7.36  6.46  125.1  100.1  6.59  5.62  4.91  4.53  66.73  66.73
 4998 µg/pl.  Mean  171  100  10  9 129   183  198  209  19  14
   sd  28  8 3 1   11  17  18  68 
   f(I)  1.23  0.92  0.77  0.82 0.85  0.99  0.93  1.09  1.27  0.88 
 1499 µg/pl.  Mean  156  146  12  9  170  160  198  178  15  18
   sd  17  29  3  3  19  30  32  54  4  3
   f(I)  1.12  1.36  0.92  0.82  1.12  0.86  0.93  0.93  1.00  1.13
 500 µg/pl.  Mean  139  133  16  8  160  152  157  176  13  15
   sd  43  10  4  2  25  59  31  51  2  4
   f(I)  1.00 1.22   1.23  0.73  1.05  0.82  0.74  0.92  0.87  0.94
 150 µg/pl.  Mean  134  113  10  9  152  178  209  168  15  12
   sd  41  7  4  2  17  37  50  45  4  4
   f(I)  0.96  1.04  0.77  0.82  1.00  0.96  0.99  0.88  1.00  0.75
 50 µg/pl.  Mean  135  145  10  6  145  127  218  200  17  20
   sd  42  34  4  2  11  26  18  54  2  5
   f(I) 0.97  1.33  0.77   0.55  0.95  0.69  1.03  1.04  1.13  1.25

In this table ">1000" is represented by "1001"

Mean Revertants Second Experiment:

Strain   97a    98 100  102    1535  
 Induction  -S9  +S9  -S9  +S9  -S9  +S9  -S9  +S9  -S9  +S9
 H2O  Mean  99  118  5  12  160  151  156  137  15  13
   sd  54.9  13.6  1.7  3.9  23.5  20.1  14.0  25.0  1.8  4.9
 DMSO Mean   152  115  7  12  142  133  167  164  8  11
   sd  9.6  6.1  0.8  3.4  17.0  1.9  8.6  31.0  2.1  2.2
 Pos.Contr.  Mean  1001  1001  1001  1001  1001  1001  1001  1001  1001  1001
   sd  0  0  0  0  0  0  0  0  0  0
   f(I)  6.59  8.70  143.0  83.42  6.26  7.53  5.99  6.10  66.73  91.00
 4998 µg/pl. Mean   126  115  8  11  112  187  141  156  10  15
   sd  35  45  4 10  46  15    27  3  5
   f(I)  1.27  0.97  1.60  0.92 0.70   1.24  0.90  1.14  0.67  1.15
 2499 µg/pl.  Mean  123  142  8  7  150  130  185  143  12  9
   sd  39  24  4  6  14  47  13  46  6  3
   f(I)  1.24  1.20  1.60  0.58  0.94  0.86  1.19  1.04  0.80  0.69
 1250 µg/pl.  Mean  145  149  10  9  164  157  204  182  15  12
   sd  10  10  5  1  5  16  9  33  5  2
  f(I)   1.46  1.26 2.00  0.75  1.03  1.04  1.31  1.33   1.00  0.92

In this table "> 1000" is represented by "1001"

Applicant's summary and conclusion

Conclusions:
CC10 Zirconium Oxide is considered as "not mutagenic under the conditions of the test."