Aluminium, compound with nickel (1:1)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Not mutagenic under the conditions of the test (S9 mix, with and without metabolic activation).

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-08-17 - 2015-09-30

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

see vehicle

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

Salmonella typhimurium LT2

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Details on mammalian cell type (if applicable):

Genetically changed strains of Salmonella typhimurium. All strains used) were obtained from TRINOVA BioChem (batch of the bacteria strains: TA97a: 4904D, TA98: 4903D, TA100: 4902D, TA102: 4872D, TA1535: 4908D) and were stored as lyophilisates in the fridge at 2-8 °C.

Metabolic activation:

with and without

Metabolic activation system:

S9-mix, rat liver S9-mix induced by Aroclor 1254

Test concentrations with justification for top dose:

First Experiment:
5 concentrations of the test item, suspended in demineralised water were used:

Nominal concentrations / Real concentrations
5000 μg/plate (non-centrifugal) / 5000 μg/plate
5000 μg/plate / resulting test item solution 100%
1500 μg/plate / resulting test item solution 30%
500 μg/plate / resulting test item solution 10%
150 μg/plate / resulting test item solution 3%
50 μg/plate / resulting test item solution 1%


Second Experiment:
To verify the results of the first experiment, a second experiment was performed, using 6 concentrations of the test item and a modification in study performance (pre-incubation method):

Nominal concentrations / Real concentrations
5000 μg/plate (non-centrifugal) / 5000 μg/plate
5000 μg/plate / resulting test item solution 100%
2500 μg/plate / resulting test item solution 50%
1250 μg/plate / resulting test item solution 25%
625 μg/plate / resulting test item solution 12.5%
313 μg/plate / resulting test item solution 6.25%
156 μg/plate / resulting test item solution 3.13%

Vehicle / solvent:

The test item is completely insoluble in demineralised water, ethanol or DMSO. Therefore, preparation of the test item according to the requirements of the OECD guideline is not possible.
In consultation with the monitor, the following procedure is considered to be the best approach:
a stock suspension of the test item in demineralised water was prepared (containing 50 g/L) and shaken for 96 h. On the day of the test, the suspension was used as highest concentration of the test item. The remaining suspension was centrifuged and the supernatant (= resulting test item solution) was used to prepare the geometric series of dilutions to be tested.

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

other: Nitrophenylendiamine, 2-Amino-anthracene

Details on test system and experimental conditions:

8 h before the start of each experiment, one lyophilisate per strain to be used was taken from the fridge to inoculate a culture vessel containing nutrient broth. After incubation overnight at 37 ± 1 °C, the cultures were used in the experiment. During the test, the cultures were stored at room temperature as to prevent changes in the titre.

Culture of Bacteria
8 h before the start of each experiment, the nutrient broth was inoculated. For the incubation of strains TA97a, TA98, TA100, TA102; ampicilline was added to
the nutrient broth (24 mg/L), for the incubation of strain TA102, tetracycline was added (2 mg/L) in addition to ampicilline. TA1535 was incubated without the addition of antibiotics. The flasks were incubated at 37 ±1°C for 8 h.

Evaluation criteria:

The colonies were counted visually, the numbers were recorded. A spreadsheet software (Microsoft Excel®) was used to calculate mean values and standard deviations of each treatment, solvent control and positive control.
The increase factor f(I) of revertant induction (mean revertants divided by mean spontaneous revertants) and the absolute number of revertants (“Rev. abs.”, mean revertants less mean spontaneous revertants) were also calculated.
A test item is considered to have mutagenic potential, if a significant, reproducible increase of revertant colonies per plate (increase factor ³ 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.

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The test item showed no precipitates on the plates in the tested concentrations with one exception: precipitates were observed in the highest non-centrifugal concentration 5000 μg/plate.
No signs of toxicity towards the bacteria (S9 mix, with and without metabolic activation) could be observed, thus the Ames test is negative:
Under the conditions of the test, the test item did not show mutagenic effects towards Salmonella typhimurium, strains TA97a, TA98, TA100, TA102 and TA1535. Therefore, no concentration-effect relationship could be determined. No significant increase of the number of revertant colonies in the treatments with and without metabolic activation could be observed.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Not mutagenic under the conditions of the test (S9 mix, with and without metabolic activation).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Justification for selection of genetic toxicity endpoint

RL1