[No public or meaningful name is available]

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

21.4. - 12.7.2021

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

2019,2020

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,1,3,3-tetrachloroprop-1-ene

EC Number:

675-808-5

Cas Number:

18611-43-3

Molecular formula:

C3H2Cl4

IUPAC Name:

1,1,3,3-tetrachloroprop-1-ene

Test material form:

liquid

Details on test material:

concentration 99,87%

Method

Target gene:

he experiments were carried out using histidine-requiring auxotroph strains of Salmonella
typhimurium (Salmonella typhimurium TA98, TA100, TA1535 and TA1537), and the
tryptophan-requiring auxotroph strain of Escherichia coli (Escherichia coli WP2 uvrA) in
the presence and absence of a post mitochondrial supernatant (S9) prepared from livers of
Phenobarbital/β-naphthoflavone-induced rats.

Metabolic activation:

with

Metabolic activation system:

The experiments were carried out using histidine-requiring auxotroph strains of Salmonella
typhimurium (Salmonella typhimurium TA98, TA100, TA1535 and TA1537), and the
tryptophan-requiring auxotroph strain of Escherichia coli (Escherichia coli WP2 uvrA) in
the presence and absence of a post mitochondrial supernatant (S9) prepared from livers of
Phenobarbital/β-naphthoflavone-induced rats.

Test concentrations with justification for top dose:

in the case of Salmonella typhimurium strains:
-S9: 1000, 500, 160, 50, 16 and 5 μg/plate,
+S9: 1600, 500, 160, 50, 16 and 5 μg/plate;
in the case of Escherichia coli WP2 uvrA:
±S9: 1600, 500, 160, 50, 16 and 5 μg/plate.

Vehicle / solvent:

dimethyl sulfoxide

Controls

Negative solvent / vehicle controls:

yes

Remarks:

Dimethyl sulfoxide (DMSO)

True negative controls:

yes

Positive controls:

yes

Positive control substance:

other:

Details on test system and experimental conditions:

The study includes a preliminary solubility test, a preliminary concentration range finding
test (informatory toxicity test) an initial mutation test (plate incorporation test) and a
confirmatory mutation test (pre-incubation test).
In the preliminary concentration range finding test as well as in the initial mutation test the
plate incorporation method was used.

A standard plate incorporation procedure was performed as an initial mutation test. Bacteria
(cultured in Nutrient Broth No.2. (Section: 5.4.2)) were exposed to the test item both in the
presence and absence of rat liver S9 as metabolic activation system.
Molten top agar was prepared and kept at 45°C. Two mL of top agar was aliquoted into
individual test tubes (3 tubes per controls or concentration level). The equivalent number of
minimal glucose agar plates was properly labelled. Conditions were investigated in triplicate.
The test item and other components were prepared fresh and added to the overlay (45°C).

Evaluation criteria:

The colony numbers on the untreated, vehicle control, positive control and the test plates
were determined visually by manual counting and the mean values, standard deviations and
the mutation rates were calculated.

An increase is considered biologically relevant if:
- in strain Salmonella typhimurium TA100 the number of reversions is at least twice as high
as the reversion rate of the vehicle control,
- in strain Salmonella typhimurium TA98, TA1535, TA1537 and Escherichia coli WP2
uvrA the number of reversions is at least three times higher than the reversion rate of the
vehicle control.

Statistics:

No precipitation of the test item was observed on the plates after about 48 hours incubation
in the examined bacterial strains at any examined concentration level (±S9) in the performed
main experiments.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with

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

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

In general, 160 μg/plate in absence exogenous metabolic activation was considered as the lowest concentration showing unequivocal cytotoxicity.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

In general, 160 μg/plate in absence (−S9), exogenous metabolic activation (+S9)) was considered as the lowest concentration showing unequivocal cytotoxicity.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

In general, 160 μg/plate in the presence of exogenous metabolic activation (+S9)) was considered as the lowest concentration showing unequivocal cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with

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

Additional information on results:

The reported data of this mutagenicity assay show that under the experimental conditions
applied, the test item did not induce gene mutations by base pair changes or frameshifts
in the genome of the strains used.

Remarks on result:

no mutagenic potential (based on QSAR/QSPR prediction)

Applicant's summary and conclusion

Conclusions:

The reported data of this mutagenicity assay show that under the experimental conditions
applied, the test item did not induce gene mutations by base pair changes or frameshifts
in the genome of the strains used.
In conclusion, the test item 1133-Tetrachloropropene has no mutagenic activity on the
applied bacterium tester strains under the test conditions used in this study.

Executive summary:

The test item 1133-Tetrachloropropene has no mutagenic activity on the
applied bacterium tester strains under the test conditions used in this study