Benzeneacetonitrile, 4-chloro-.alpha.-[(3-chloro-2-fluorophenyl)methylene]-2-fluoro-, (.alpha.Z)-

Genetic toxicity: in vitro

Administrative data

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:

20 October 2015 - 14 January 2016

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(2Z)-3-(3-chloro-2-fluorophenyl)-2-(4-chloro-2-fluorophenyl)prop-2-enenitrile

EC Number:

806-533-2

Cas Number:

1219086-87-9

Molecular formula:

C15 H7 Cl2 F2 N

IUPAC Name:

(2Z)-3-(3-chloro-2-fluorophenyl)-2-(4-chloro-2-fluorophenyl)prop-2-enenitrile

Test material form:

other: white solid

Details on test material:

Roche number : RO5463921-000
Batch : FIS201405080058
Molecular mass : 310,3 g/mol
Purity : 101,2 % (m/m)
Expiry date : 11 June 2016
Storage conditions : at room temperature, light protected

Method

Target gene:

- S. typhimurium: Histidine gene
- E. coli: Tryptophan gene

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

Properly maintained: yes

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

Properly maintained: yes

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

The test item was tested at the following concentrations at both experiments :
3; 10; 33; 100; 333; 1000; 2500 and 5000 µg/plate

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

withour activation

Controls 2

Positive control substance:

other: 4-nitro-o-phenylene-diamine

Remarks:

without activation

Controls 3

Positive control substance:

methylmethanesulfonate

Remarks:

without activation

Controls 4

Positive control substance:

other: 2-aminoanthracene

Remarks:

with activation

Results and discussion

Test results

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:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported according to OECD 471, the PAC- Ethenenitrile did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

Executive summary:

This study was performed to investigate the potential of PAC- Ethenenitrile (CAS Nr. 1219086-87-9) to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and the Escherichia coli strain WP2 uvrA according to OECD 471.

The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations in both experiments: 3; 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate.

The test item precipitated in the overlay agar in the test tubes from 333 to 5000 µg/plate in the absence of S9 mix and from 100 to 5000 µg/plate in the presence of S9 mix. Precipitation of the test item in the overlay agar on the incubated agar plates was observed from 333 to 5000 µg/plate in experiment I without S9 mix and in experiment II with and without S9 mix. In experiment I with S9 mix precipitation was observed from 100 to 5000 µg/plate. The undissolved particles had no influence on the data recording.

The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without S9 mix in all strains used.

Toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in nearly all strains.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with PAC- Ethenenitrile at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no dose dependent increase in the number of revertant colonies below the threshold.

Appropriate reference mutagens were used as positive controls. They showed a distinct increase of induced revertant colonies. Negative and solvent controls showed regular background growth and mean revertant colony numbers in the range of the historical data.

Therefore the test was considered valid.