Sodium 2-amino-4,6-dinitrophenoxide

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro Mammalian gene mutation assay

Under the experimental conditions reported, the test item did not induce mutations in the mouse lymphoma thymidine kinase locus assay using the cell line L5178Y in the absence and presence of metabolic activation.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from secondary literature.

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Principles of method if other than guideline:

To evaluate the mutagenic potential of test chemical in mouse lymphoma L5178Y cells by In mammalian cell gene mutation assay.

GLP compliance:

not specified

Type of assay:

mammalian cell gene mutation assay

Species / strain / cell type:

mouse lymphoma L5178Y cells

Additional strain / cell type characteristics:

other: tk+/-

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

experiment 1: 112.5, 225, 450, 900, 1350 and 1800 μg/ml with and without S9-mix
experiment 2: 28.1, 56.3, 112.5, 225, 337.5 and 450 μg/ml without S9-mix
112.5, 225, 450, 750, 900, 1050 and 1200 μg/ml with S9-mix

Vehicle / solvent:

de-ionised water

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

cyclophosphamide

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Details on test system and experimental conditions:

Treatment experiment 1: 4h treatment with and without S9-mix
experiment 2: 4h treatment with and 24h without S9-mix

Evaluation criteria:

The cells were evaluated for mutagenic potential.

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: No mutagenic effect were observed

Conclusions:

Under the experimental conditions reported, the test item did not induce mutations in the mouse lymphoma thymidine kinase locus assay using the cell line L5178Y in the absence and presence of metabolic activation.

Executive summary:

Test substance was assessed for its possible mutagenic potential. For this purpose In vitro Mammalian cell gene mutation assay was performed as per OECD 476 in Mouse Lymphoma cell line L5178Y. The test material was exposed at the concentration mention below.

experiment 1: 112.5, 225, 450, 900, 1350 and 1800 μg/ml with and without S9-mix

experiment 2: 28.1, 56.3, 112.5, 225, 337.5 and 450 μg/ml without S9-mix

                         112.5, 225, 450, 750, 900, 1050 and 1200 μg/ml with S9-mix

 

 

No substantial and reproducible concentration-dependent increase in mutant colony numbers was observed in both main experiments. No relevant shift of the ratio of small versus large colonies was observed up to the maximal concentration of the test item. Appropriate reference mutagens were used as positive controls and showed a distinct increase in induced mutant colonies, indicating that the tests were sensitive and valid. The concentration range of the main experiments was adjusted to toxicity data and the occurrence of precipitation. Therefore test substance was considered to be non-mutagenic in Mouse Lymphoma cell line L5178Y by In vitro Mammalian cell gene mutation assay. Hence the substance cannot be classified as gene mutant in vitro.

Endpoint conclusion

Endpoint conclusion:

no study available

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Data for the various test chemicals was reviewed to determine the mutagenic nature of sodium 2-amino-4,6-dinitrophenolate ( 831-52-7). The studies are as mentioned below:

AMES test

This study was performed to assess the mutagenic potential of test substance to induce gene mutations in comparison to negative control according to the plate incorporation method (Trial-I) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102. The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the negative and positive controls was tested in triplicates. The test item was tested at the following concentrations i.e., 0.0 (NC),0.050, 0.158, 0.501, 1.582 and 5 mg/plate, both in presence (+S9) and in absence (-S9) of metabolic activation. A significant increase in revertant colony numbers were observed in the tester strains (TA 98 and TA 1537) following treatment with test substance both in the presence and absence of metabolic activation (S9 mix) except TA 1535, TA 100 and TA 102. High mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance were observed. The spontaneous reversion rates in the negative and positive controls were within the range of our historical data.Reference mutagens showed a distinct in­crease in induced revertant colonies in TA98 and TA1537 strains both in the presence (+S9) as well as in the absence (-S9) of metabolic activation, without showing any signs of cytotoxicity.The positive controls used for various strains showed a distinct increase in induced revertant colonies in the Plate incorporation method.

Conclusion

It is concluded that the test substance did not induce gene mutations by frame shifts in the genome of the Salmonella typhimurium strains TA 1535, TA 100 and TA 102 in the presence and absence of S9 metabolic activation system while there were mutagenic effects were observed in TA 98 and TA 1537 both in the presence and absence of metabolic activation.

 

In vitro Mammalian gene mutation assay

Test substance was assessed for its possible mutagenic potential. For this purpose In vitro Mammalian cell gene mutation assay was performed as per OECD 476 in Mouse Lymphoma cell line L5178Y. The test material was exposed at the concentration mention below.

experiment 1: 112.5, 225, 450, 900, 1350 and 1800 μg/ml with and without S9-mix

experiment 2: 28.1, 56.3, 112.5, 225, 337.5 and 450 μg/ml without S9-mix

                         112.5, 225, 450, 750, 900, 1050 and 1200 μg/ml with S9-mix

 

 

No substantial and reproducible concentration-dependent increase in mutant colony numbers was observed in both main experiments. No relevant shift of the ratio of small versus large colonies was observed up to the maximal concentration of the test item. Appropriate reference mutagens were used as positive controls and showed a distinct increase in induced mutant colonies, indicating that the tests were sensitive and valid. The concentration range of the main experiments was adjusted to toxicity data and the occurrence of precipitation. Therefore test substance was considered to be non-mutagenic in Mouse Lymphoma cell line L5178Y by In vitro Mammalian cell gene mutation assay. Hence the substance cannot be classified as gene mutant in vitro.

 

Based on the data summarized, sodium 2-amino-4,6-dinitrophenolate ( 831-52-7) was mutagenic in two strain of Salmonella typhimurium TA 98 and TA 1537 both in the presence and absence of metabolic activation. While the test substance exhibit a negative result in confirmatory test i.e. inIn vitro Mammalian gene mutation assay. The test substance was considered to be non-mutagenic in Mouse Lymphoma cell line L5178Y both in the presence and absence of metabolic activation. Hence the substance cannot be classified as gene mutant in vitro. .Hence it is not likely to be mutagenic in vitro.

Justification for classification or non-classification

Thus based on the above annotation and CLP criteria the test chemical sodium sodium 2-amino-4,6-dinitrophenolate ( 831-52-7) did not induce gene mutation in confirmatory test i.e. in In vitro Mammalian gene mutation assay and 3 strains of Salmonella typhimurium .Hence it is not likely to be classified as mutagenic in vitro.