1,3-Diazaspiro[4.5]decane-2,4-dione, 8-methoxy-, sodium salt (1:1), cis-

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation (bacterial reverse mutation assay / Ames test, GLP, OECD TG471): negative with and without metabolic activation

Chrom. Abb. (clastogenicity mammalian cells / GLP, OECD TG473): negative with and without metabolic activation

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 16 - November 09, 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Version / remarks:

1997

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

liver S9 mix of Aroclor 1254 induced rats

Test concentrations with justification for top dose:

Pre-test: 0, 10, 50, 100, 200, 400, 800, 1600 and 2400 for 4 and 18 hours treatment
Main study: 0, 600, 1200 and 2400 µg/ml for 4 hours treatment

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: mitomycin C, cyclophosamide

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

Based on this test, the test item is considered not to be clastogenic for mammalian cells in vitro.

Executive summary:

The clastogenic potential of the test item was evaluated in a chromosome aberration test in vitro. Initially Chinese hamster V79 cells were exposed in the absence and in the presence of S9 mix for 4 hours to concentrations of 600, 1200 and 2400 ug/ml.

Without S9 mix cytotoxic effects were not observed after 4 hours treatment and after 18 hours treatment. With S9 mix cytotoxic effects were observed at 2400 ug/ml. Precipitation in the medium did not occur. However, the test item  was tested up to and over the requested limit concentration of 10mM (equal to 2202 ug/ml).
None of the cultures treated with the test item in the absence and in the presence of S9 mix showed biologically relevant or statistically significant increased numbers of aberrant metaphases.
The positive controls mitomycin C and cyclophosphamide induced clastogenic effects and demonstrated the sensitivity of the test system and the activity of the used S9 mix.
Based on this test, the test item is considered not to be clastogenic for mammalian cells in vitro.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 05 - October 30, 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

no

Remarks:

according to current version (2020) no bacteria strain included to detect cross-linking mutagens

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

liver S9 mix of Aroclor 1254 incuded rats

Test concentrations with justification for top dose:

16, 50, 158, 500, 1581 and 5000 µg/plate

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide, nitrofurantoin, 4-nitro-1,2-phenylene diamine, mitomycin C, cumene hydroperoxide and 2-aminoanthracene

Key result

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

The test substance was considered to be non-mutagenic without and with S9 mix in the plate incorporation as well as in the preincubation modification of the Salmonella/microsome test.

Executive summary:

 

The test substance was tested in the Salmonella/microsome test for point-mutagenic effects in doses up to 5000 µg/plate on the five histidine-auxotrophic Salmonella typhimurium LT2 strains TA 1535, TA 100, TA 1537, TA 1538 TA 98 and TA 102.

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the results there is no classification required according to Regulation (EC) 1272/2008 (CLP).