2-methyl-1,4-phenylene-bis[4-(6-acryloyloxyhexyloxy)benzoate]

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:

May 10 to May 18, 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

August 1983

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Version / remarks:

August 1983

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 1992

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:

-

EC Number:

419-310-6

EC Name:

-

Cas Number:

125248-71-7

Molecular formula:

C39H44O10

IUPAC Name:

2-methyl-4-(4-{[6-(prop-2-enoyloxy)hexyl]oxy}benzoyloxy)phenyl 4-{[6-(prop-2-enoyloxy)hexyl]oxy}benzoate

Test material form:

solid

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Liver S9-Mix from Aroclor 1254-pretreated rats

Test concentrations with justification for top dose:

First experiment: 0.5, 1.58, 5, 15.8, 50, 158, and 500 µg/plate
Second experiment: 5, 15.8, 50, 158, and 500 µg/plate

The highest concentration of 500 µg/plate was chosen as higher concentrations were not soluble in the solvent.

Vehicle / solvent:

- Vehicle/solvent used: acetone
- Justification for choice of solvent/vehicle: Historical data of the laboratory and experience of other research groups (Maron et al. 1981) showed that such amounts of the selected solvents have no influence on the number of spontaneous revertants of any strain.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

cumene hydroperoxide

other: 2-Aminoanthracene, Daunomycin

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 - 72 hours
-
NUMBER OF REPLICATIONS: three

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of spontaneous revertants, clearing of background lawn of non-revertant bacteria

Evaluation criteria:

A test material is defined as non-mutagenic in this assay if
- "no" or "weak increases" occur in the first and second series of the main experiment.
("Weak increases" randomly occur due to experimental variation.)

A test material is defined as mutagenic in this assay if
- a dose-related (over at least two test material concentrations) increase in the number of revertants is induced, the maximal effect is a "clear increase", and the effects are reproduced at similar concentration levels in the same test system;

- "clear increases" occur at least at one test material concentration, higher concentrations show streng precipitation or cytotoxicity, and the effects are reproduced at the same concentration level in the same test system.

In all further cases should a third test series with the bacterial strain in question be performed. lf the criteria for a positive test result are not fulfilled in at least two out of the three series, the test material is defined as being non-mutagenic in this test system.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation occurred at the highest contration tested 500 µg/plate.

RANGE-FINDING/SCREENING STUDIES: The test material was investigated in the first experimental series at seven concentrations, separated by half-log intervals, ranging from 0.5 to 500 μg per plate. Higher concentrations were not soluble in the solvent used. In the second experimental series, usually 5 concentrations including at least 4 non-toxic concentrations should be tested.

Applicant's summary and conclusion

Conclusions:

With and without addition of S9-Mix as the external metabolizing system, the test item was not mutagenic under the experimental conditions described.

Executive summary:

The investigations for mutagenic potential were performed using Salmonella typhimurium TA 98, TA 100, TA 102, TA 1535, and TA 1537 as tester strains. The plate incorporation test with and without addition of liver S9-Mix from Aroclor 1254- pretreated rats was used.

The test material was tested in two series of experiments at concentrations ranging from 0.5 to 500 μg/plate. Higher concentrations were not soluble in the solvent used. Precipitation of the test material on the agar plates was macroscopically visible at the highest concentration used (500 μg/plate). Toxicity to the bacteria was not observed.

Daunomycin, N-ethyl-N'-nitro-N-nitrosoguanidine, 9-aminoacridine and cumene hydroperoxide served as strain specific positive control compounds in the absence of S9-Mix. 2-Aminoanthracene and benzo[a]pyrene were used for testing the bacteria and the activity of the S9-Mix. Each treatment with the substances used as positive controls led to a clear increase in revertant colonies, thus showing the expected reversion properties of all strains and good metabolic activity of the S9-Mix used. With and without addition of S9-Mix as the external metabolizing system, the test material was not mutagenic under the experimental conditions described.