4,4'-bis(2-methoxystyryl)-1,1'-biphenyl

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 21. Jul. 1997

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

adopted 30. May 2008

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-bis(2-methoxystyryl)-1,1'-biphenyl

EC Number:

254-935-6

EC Name:

4,4'-bis(2-methoxystyryl)-1,1'-biphenyl

Cas Number:

40470-68-6

Molecular formula:

C30H26O2

IUPAC Name:

4,4'-bis(2-methoxystyryl)-1,1'-biphenyl

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material: 4,4'-bis(2-methoxystyryl)-1,1'-biphenyl
- IUPAC name: 1-[(E)-2-(2-methoxyphenyl)ethenyl]-4-{4-[(E)-2-(2-methoxyphenyl)ethenyl]phenyl}benzene
- Molecular formula: C30H26O2
- Molecular weight: 418.53 g/mol
- Smiles notation: COC1=CC=CC=C1/C=C/C2=CC=C(C3=CC=C(/C=C/C4=CC=CC=C4OC)C=C3)C=C2
- InChl: 1S/C30H26O2/c1-31-29-9-5-3-7-27(29)21-15-23-11-17-25(18-12-23)26-19-13-24(14-20-26)16-22-28-8-4-6-10-30(28)32-2/h3-22H,1-2H3/b21-15+,22-16+
- Substance type: Organic
- Physical state: Solid

Method

Species / strain

Species / strain 1

Species / strain / cell type:

other: S. typhimurium TA 97a

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 98

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 100

Additional strain / cell type characteristics:

not specified

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 102

Additional strain / cell type characteristics:

not specified

Species / strain 5

Species / strain / cell type:

S. typhimurium TA 1535

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

metabolic activation by microsomal enzymes (e.g., benzo(a)pyrene)

Test concentrations with justification for top dose:

5000 µg/plate

Vehicle / solvent:

vehicle

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

other: 4-Nitro-1,2-phenylene diamine, C6H7N3O2; CAS-No.: 99-56-9 and 2-Amino-anthracene, C14H11N; CAS-No.: 613-13-8

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension

DURATION
- Preincubation period: 8 hours at 37 ± 1 °C
- Exposure duration: First Experiment (72 hours at 37 ±1 °C), Second Experiment (48 hours at 37 ±1 °C)
- Expression time (cells in growth medium): 24 hours at 37 ±1 °C
- Selection time (if incubation with a selection agent): 48 hours at 37 ±1 °C
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hours at 37 ±1 °C

SELECTION AGENT (mutation assays): mutagenic substances: 4-Nitro-1,2-phenylene diamine (CAS#99-56-9), Sodium azide (CAS#26628-22-8), 2-Amino-anthracene(CAS#613-13-8) and Benzo-a-pyrene(CAS#50-32-8)

NUMBER OF REPLICATIONS: 3

Rationale for test conditions:

Nearly all negative and all strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Evaluation criteria:

The determination of titre should give a number of at least 109 cells/mL

The determination values for the spontaneous revertants of the negative controls and positive controls (diagnostic mutagens) were within the historical control data ranges.

Statistics:

The confirmation tests of the genotype did not show any irregularities. The control of the titre was above the demanded value of 109 bacteria/mL. Nearly all of the means of all replicates of the spontaneous revertants (in negative and solvent controls) were within the range of the historical data of the test facility. All numbers of revertant colonies of the positive controls were within the range of the historical data of the laboratory and were increased in comparison with the negative controls, which demonstrated the mutagenic potential of the diagnostic mutagens.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

other: S. typhimurium TA 97

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

Test results 2

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

Positive controls validity:

valid

Test results 3

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

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium 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

Test results 5

Key result

Species / strain:

S. typhimurium TA 1535

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

Applicant's summary and conclusion

Conclusions:

Based on the results of this study it is concluded that 4,4'-bis(2-methoxystyryl)-1,1'-biphenyl is not mutagenic in the Salmonella typhimurium strains TA97a, TA98, TA100, TA102 and TA1535 in the absence and presence of metabolic activation un-der the experimental conditions in this study.

Executive summary:

Two valid experiments were performed.

 

The study procedures described in this report were based on the most recent OECD and EC guidelines.

The test item 4,4'-bis(2-methoxystyryl)-1,1'-biphenyl was tested in the Salmonella typhimurium reverse mutation assay with five strains of Salmonella typhimurium (TA97a, TA98, TA100, TA102 and TA1535).

The test was performed in two experiments in the presence and absence of metabolic activation, with +S9 standing for presence of metabolic activation, and –S9 standing for absence of metabolic activation.

 

Experiment 1:

In the first experiment,the test item (suspended in ethanol) was tested up to concentrations of 5000 µg/plate in the absence and presence of S9-mix in the strains TA97a, TA98, TA100, TA102 and TA1535 using the plate incorporation method.

The test item showed precipitates on the plates at the highest concentration (5000 µg/plate), only.

The bacterial background lawn was not reduced at any of the concentrations and no relevant decrease in the number of revertants was observed in all bacteria strains. The test itemshowed no signs of toxicity towards the bacteria strains in both the absence and presence of metabolic activation.

The results of this experiment showed that none of the tested concentrations showed a significant increase in the number of revertants in all tested strains, in the presence and the absence of metabolic activation.

 

Experiment 2:

Based on the first experiment,the test item was tested up to concentrations of 5000 µg/plate in the absence and presence of S9-mix in all bacteria strains using the pre-incubation method.

The test item showed precipitates on the plates at the two highest concentrations (5000 and 2500 µg/plate).

The bacterial background lawn was not reduced at any of the concentrations and no relevant decrease in the number of revertants was observed in all bacteria strains. The test itemshowed no signs of toxicity towards the bacteria strains in both the absence and presence of metabolic activation.

The results of this experiments showed that the test item caused no increase in the number of revertants in all bacteria strains compared to the solvent control, in both the absence and presence of metabolic activation. The test item did not induce a dose-related increase in the number of revertants colonies in all strains, in the presence and absence of metabolic activation.