Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Bacterial gene mutation: The test material is non-mutagenic to bacterial cells (S. typhimurium and E. Coli). In an additional read across study, a similar substance is non-mutagenic to bacterial cells (S. typhimurium). Cytogenicity: In a read across study a similar substance is non-clastogenic to human lymphocytes. In an additional read across study is non-clastogenic to rat lymphocytes. Gene mutation in mammalian cells: In a read across study, is non-mutagenic to mouse lymphoma.

Link to relevant study records
Reference
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:
April to June 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according 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
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
Not applicable
Species / strain / cell type:
E. coli WP2 uvr A
Details on mammalian cell type (if applicable):
Not applicable
Metabolic activation:
with and without
Metabolic activation system:
S-9
Test concentrations with justification for top dose:
Experiment 1: 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate.
Experiment 2: 50, 150, 500, 1500 and 5000 µg/plate.
Vehicle / solvent:
Vehicle(s)/solvent(s) used: Acetone
Justification for choice of solvent/vehicle: The test material was initially noted to be insoluble in sterile distilled water, dimethyl sulphoxide, dimethyl formamide and acetonitrile at 50 mg/mL and acetone at 100 mg/mL but was fully soluble in tetrahydrofuran at 200 mg/mL. However, the test material in tetrahydrofuran could not be adequately dosed because the formulation immediately solidified during the pipetting stage. Therefore, the most suitable suspension was selected. The test material formed the best doseable suspension in acetone, therefore, this solvent was selected as the vehicle.
Untreated negative controls:
yes
Remarks:
ENNG: 2µg/plate for WP2uvrA, 3µg/plate for TA100, 5µg/plate for TA1535; 9AA: 80µg/plate for TA1537; 4NQO: 0.2µg/plate for TA98.
Negative solvent / vehicle controls:
yes
Remarks:
Acetone
True negative controls:
not specified
Positive controls:
yes
Remarks:
2AA: 1µg/plate for TA100, 2µg/plate for TA1535 and TA1537, 10µg/plate for WP2uvrA; BP: 5µg/plate for TA98.
Positive control substance:
benzo(a)pyrene
other: 2-Aminoanthracene (2AA)
Details on test system and experimental conditions:
METHOD OF APPLICATION:
Acetone is toxic to the bacterial cells at 100µl after employing the pre-incubation modification, therefore all of the formulation for Experiment 2 were prepared at concentrations two times greater than required on Vogel-Bonner agar plates. To compensate, each formulation was dosed using 0.05 mL (50µL) aliquots.
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results:
negative

The test material was considered to be non-mutagenic under the conditions of the study.
Executive summary:

In an OECD 471 study, conducted according to GLP, the test material is non-mutagenic (negative) to Salmonella typhimurium and Escherichia coli bacterial strains.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

The registration of the test material employs a combined approach for addressing the required endpoints. When data are not available, data from the read across substances are used to address the required endpoints.

 

The read across substances are structurally similar to registered substance. The substance and the proposed structural analogues are predominantly C32 – C36 alkyl fatty amides. These substances are expected to be metabolised by fatty acid amide hydrolase and esterases. Any differences among the substance structures are not expected to result in any significant differences in the toxicological effects or degradation products.

 

The following endpoints include supporting studies to provide evidence of read across suitability

  •  In vitro gene mutation study in bacterial cells;
  • In vitro cytogenicity study in mammalian cells; and
  • In vitro gene mutation study in mammalian cells.

 

Bacterial gene mutation:

Key Study:

In an OECD 471 study, conducted according to GLP, the test material is non-mutagenic (negative) to Salmonella typhimurium and Escherichia coli bacterial strains.

Supporting Study:

In an OECD 471 study, conducted according to GLP, the structurally similar read across substance is non-mutagenic to Salmonella typhimurium bacterial cells.

 

Cytogenicity:

Key Study:

In an OECD 473 study, conducted according to GLP, the structurally similar read across substance is non-clastogenic to human lymphocytes.

Supporting Study:

In an OECD 473 study, conducted according to GLP, the structurally similar read across substance  is non-mutagenic to rat lymphocytes.

 

Gene mutation in mammalian cells:

In an OECD 476 study, conducted according to GLP, the structurally similar read across substance is non-mutagenic to mouse lymphoma.

 


Justification for selection of genetic toxicity endpoint
Study conducted according to OECD under GLP conditions. Available genetic toxicity study on the registered substance, additional endpoints conducted on read-across substances

Justification for classification or non-classification

In a bacterial mutagenicity study, the test material is non-mutagenic (negative) to Salmonella typhimurium and Escherichia coli bacterial strains. According to Regulation (EC) No 1272/2008 on the classification, labelling and packaging of substances and mixtures (CLP), a substance is classified as a mutagenic if the result is positive. The test material is, therefore, not mutagenic.