Dodecanedioic acid, 1,12-bis[2-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)-3-hydroxyphenoxy]ethyl] ester

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:

12th April 2011 - 28th April 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

1,12-bis({2-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)-3-hydroxyphenoxy]ethyl}) dodecanedioate

EC Number:

939-042-8

Cas Number:

1482217-03-7

Molecular formula:

C58H56N6O8

IUPAC Name:

1,12-bis({2-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)-3-hydroxyphenoxy]ethyl}) dodecanedioate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): T-1620L
- Molecular weight (if other than submission substance): 965.10
- Physical state: pale yellow powder
- Analytical purity: 99.35%
- Lot/batch No.: 101015
- Stability under test conditions: stable at ordinary temperature and pressure
- Storage condition of test material: RT in the dark
- Other:

Specific details on test material used for the study:

- Description at room temperature: Pale yellow powder
- Purity: 99.35%

Method

Species / strain

Species / strain 1

Species / strain / cell type:

E. coli WP2 uvr A

Species / strain 2

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S9 rat liver homogenate supernatant fraction

Test concentrations with justification for top dose:

5, 20, 78, 313, 1250 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: From a solubility study conducted using distilled water, DMSO, acetone and THF, DMSO was the most suitable as it gave a suspension in which the test substance was evenly distributed. DMSO also affected the tester strains to a lesser extent than the other solvents.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

benzo(a)pyrene

furylfuramide

other: 2-aminoanthracene

Details on test system and experimental conditions:

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

DURATION
- Preincubation period: 20mins at 37°C
- Exposure duration: 48h at 37° C
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays): revertant colonies

NUMBER OF REPLICATIONS: 3 agar plates per dose level or control (negative and positive)

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- presence of revertant colonies

OTHER EXAMINATIONS:
- antibacterial activity and precipitation

OTHER:

Evaluation criteria:

The revertant colony count obtained with each plate treated with the negative control (solvent) and positive control substances was compared with the background data and reference values determined based on the background data (mean±2 x standard deviation). Test results were regarded as having been obtained as a result of appropriate operations conducted using appropriate tester strains if revertant colony counts obtained were within the range of reference values or if their deviation from the range of reference values was considered to be due to any incidental factor based on the result of comparison with background data.
Criteria for acceptance of test results
Test results were regarded as having been obtained as a result of appropriate operations conducted using appropriate tester strains if they met the following conditions:
1) The sterility test showed no evidence of bacterial contamination.
2) Values obtained with the negative control (solvent) met the criteria for acceptance.
3) Values obtained with the positive control substances met the criteria for acceptance and were at least 2 times greater than the values obtained with the negative control (solvent).

Statistics:

Dunnett’s multiple compassion (one-sided) and linear regression analysis were conducted to assess the results obtained. Test substance-treated revertant colony counts were compared with negative control (solvent)-treated revertant colony counts, and whether differences were significant was analyzed by multiple comparison (p<0.01). If differences were found significant, dose responses were analyzed by linear regression analysis (p<0.01).

Results and discussion

Test results

Test results 1

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

Test results 2

Species / strain:

E. coli WP2 uvr A

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

Additional information on results:

The test substance did not statistically significantly increase revertant colony counts of any tester strain regardless of the dose and the presence or absence of metabolic activation. Results obtained in the main study and the confirmatory study showed reproducibility. Based on these findings, the test substance was thought to have caused no biologically significant increases attributable to the induction of mutation. Results of the sterility tests conducted with Nutrient Broth No. 2 used in pre-incubation and the test substance solution of the highest concentration used in studies, S9Mix, and 0.1M phosphate buffer solution excluded the possibility of bacterial contamination. Revertant colony counts obtained with the negative control (solvent) and positive control substances were within the range of reference values determined based on background data. In addition, all substances used as positive control (2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, sodium azide, 9-aminoacridine, 2-aminoanthracene, and benzo[α]pyrene) increased revertant colony counts at least twice as much as the negative control (solvent). All these findings obtained supported that the studies were conducted in an acceptable manner. There was no environmental factor that may have adversely affected study reliability or deviation from the protocol.

Based on these findings, the test substance was judged to show no reverse mutagenic potential under the conditions of the present study.
The test substance showed no antibacterial activity against any tester strain regardless of the presence or absence of metabolic activation. It showed precipitation at 78 μg/plate or higher doses in both the presence and absence of metabolic activation.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

The test substance failed to statistically significantly increase the revertant colony count regardless of the strain, the dose and the presence or absence of metabolic activation. In addition, test results were reproducible in the main and confirmatory studies. Based on these findings, the test substance was not regarded as having caused biologically significant increases attributable to its mutagenicity in the main study.

2. All revertant colony counts obtained with the negative control (solvent) and positive controls were acceptable in comparison with the background data. All positive control substances (2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, sodium azide, 9-aminoacridine, 2-aminoanthracene, and benzo[α]pyrene) caused 2-fold or greater increases in revertant colony counts compared to the negative control (solvent). All these findings indicate that the studies were appropriately conducted.

3. Based on these findings, it was concluded that T-16201L does not show any reverse mutagenicity under the conditions of the present study.

Executive summary:

It was concluded that T-1620L does not show any reverse mutagenicity under the conditions of the present study.