N,N'-[bicyclo[2.2.1]heptane-2,6-diylbis(methylene)]bis(2-cyanoacetamide)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

6th December 2018 to 31 December 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Please refer to the Read Across Justification document enclosed in Chapter 13 for further details.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

LME 11311
Batch Ua05 (258)
Purity 99%
Yellow Orange Solid

Target gene:

histidine synthesis

Species / strain / cell type:

S. typhimurium TA 98

Species / strain / cell type:

S. typhimurium TA 100

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9 liver homogenage purchased from MolTox Boone, NC

Test concentrations with justification for top dose:

1000, 300, 100, 30, 10, 3, 1 and 1.3 ug/well

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

sodium azide

other: 2-aminoanthracene

Details on test system and experimental conditions:

The test article was prepared in dimethyl sulfoxide (DMSO) and tested via the plate incorporation method at eight dose levels.
The tester strains will include the S. typhimurium histidine auxotrophs TA98 and TA I 00 as described by Ames er al. (l975). The S. typhimurium tester strains were from Dr. Bruce Ames, University of California, Berkeley.

Each tester strain culture will be inoculated from the appropriate frozen stock,
lyophilized pellet(s), or master plate. To ensure that cultures are harvested in late log
phase. the length of incubation will be controlled and monitored. Each inoculated
flask will be placed in a shaker/incubator programmed to begin shaking at 125 to
175 rpm and incubating at 37±2°C.
All cultures will be harvested by spectrophotometric monitoring of culture turbidity
rather than by duration of incubation since overgrowth of cultures can cause loss of
sensitivity to some mutagens. Cultures will be removed from incubation at a density
of approx imatcly l 09 cclls/m L.

Key result

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:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Key result

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:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

Under the condictions of the study, the results of the Bacterial Mutagenicity Screening Assay were concluded to be negative.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

27th November 2018 to 12th December 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Please refer to the read across justification document enclosed in Chapter 13 for further details.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

LME 11194
Batch Xbm06
Purity 99%
Whitish yellow orange solid

Target gene:

histidine synthesis

Species / strain / cell type:

S. typhimurium TA 98

Species / strain / cell type:

S. typhimurium TA 100

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9 liver homogenage purchased from MolTox Boone, NC

Test concentrations with justification for top dose:

1000, 300, 100, 30, 10, 3, 1 and 0.3 ug/well

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

sodium azide

other: 2-aminoanthracene

Details on test system and experimental conditions:

The test article was prepared in dimethyl sulfoxide (DMSO) and tested via the plate incorporation method at eight dose levels.
The tester strains will include the S. typhimurium histidine auxotrophs TA98 and TA I 00 as described by Ames er al. (l975). The S. typhimurium tester strains were from Dr. Bruce Ames, University of California, Berkeley.

Each tester strain culture will be inoculated from the appropriate frozen stock,
lyophilized pellet(s), or master plate. To ensure that cultures are harvested in late log
phase. the length of incubation will be controlled and monitored. Each inoculated
flask will be placed in a shaker/incubator programmed to begin shaking at 125 to
175 rpm and incubating at 37±2°C.
All cultures will be harvested by spectrophotometric monitoring of culture turbidity
rather than by duration of incubation since overgrowth of cultures can cause loss of
sensitivity to some mutagens. Cultures will be removed from incubation at a density
of approx imatcly l 09 cclls/m L.

Key result

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:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Key result

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:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

Under the condictions of the study, the results of the Bacterial Mutagenicity Screening Assay were concluded to be negative.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Justification for classification or non-classification

Based on testing conducted on similar substances, no classification or labelling is neccessary. Further information on the Read Across is available in the Read Across justification document enclosed in Chapter 13.