Oxybis(2,1-ethanediyloxy-2,1-ethanediyl)diacrylate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

An Ames test was available and showed no mutagenic activity in bacteria with Tetraethylene Glycol Diacrylate.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February / March 1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

The test compound was examined for mutagenic activity in a series of in vitro microbial assays employing Salmonella and Saccharomyces indicator organisms. The compound was tested directly and in the presence of liver microsomal enzyme preparations from Aroclor-induced rats.
The compound was tested over a series of concentrations such that there was evidence of sme chemically-induced physiological effects at the high dose level. The low dose in all cases was below the concentration that demonstrated any toxic effect.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Target gene:

not applicable

Species / strain / cell type:

Saccharomyces cerevisiae

Species / strain / cell type:

S. typhimurium, other: TA1535, TA1537, TA1538, TA98, TA100

Metabolic activation:

with and without

Metabolic activation system:

liver rat (Aroclor induced)

Test concentrations with justification for top dose:

-TA1535, TA1537, TA100, c.c. with and without S9: 0.01, 0.1, 1, 5 µl per plate.
-TA1538, TA98 without S9 : 0.001, 0.005, 0.01, 0.1 µl
-TA1538, TA98 with S9 : 0.01, 0.1, 1, 5 µl per plate.

Vehicle / solvent:

not specified

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

2-nitrofluorene

N-dimethylnitrosamine

other: methylnitrosoguanidine & quinacrine mustard (without S9), 2-anthramine, 8-aminoquinoline (with S9)

Details on test system and experimental conditions:

Plate test data consist of direct revertant colony counts obtained from a set of selective agar plates seeded with populations of mutan cells is suspended in a semi solid overlay. Because the test chemical and the cells were incubated in the overlay for 2 to 3 days, and a few cell divisions occur during the incubation period, the test is semi quantitative in nature.

Plate test procedures do not permit exact quantification of the number of cells surviving chemical treatment. At low concentrations of the test chemical, the surviving population on the treatment plates is essentially the same as that on the negative control plate. At high concentrations, the surviving population is usually reducedby some fraction. The protocol normally employs several doses ranging over two or three log concentrations, the highest of these doses being selected to show slight toxicity as determined by subjective criteria.
The demonstration of dose-related increases in mutant counts is an iportant criterion in establishing mutagenicity. A factor that might modify dose-response results for a mutagen would be the selection of doses that are too low (usually mutagenicity and toxicity are related). If the highest dose is far lower than a toxic concentration, no increases may be observed over the dose range selected. Conversely, if the lowest dose emplyed if highly cytotoxic, the test chemical may kill any mutants that are induced, and the copound will not appear to be mutagenic.

Positive and negative control assays are conducted with each experiment and consist of direct-acting mutagens for nonactivation assays and mutagens that require metabolic biotransformation in activation assays. negative controls consist of the test compound solvent in the overlay agar together with the other essential coponents. The negative control plate for each strain gives a reference point to which the test data are compared. The positive control assay is conducted to demonstrate that the test systems are functional with known mutagens.

Tissue homogenates and supernatants: The tissue homogenates and 9000 x g supernatants were prepared from the livers of SD adult male rats. The animals were pretreated with Arochlor 1254 (500 mg/kg) 5 days before kill.

Approximately 10^9 cells from a log phase culture of each indicator strain were added to test tubes containing 2.0 ml of molten agar supplemented with biotin and a trace of histidine. For non-activation tests, the four dose levels of the test compound were added to the contents of the appropriate tubes. In activation tests the 9000 x g tissues supernatant and required cofactors were added to the overlay tubes. The plates were incubated for 48 to 72 hours at 37°C, and scored for the number of colonies growing on each plate.

Evaluation criteria:

The demonstration of dose-related increases in mutant counts is the most reliable method to demonstrate mutagenicity. Mutant increases at only one or two doses may be significant if they occur at the higher doses. Increases at low or intermediate concentrations followed by reduced mutant counts at higher doses or that the high dose levels were toxic and the induced revertant cells were killed.
It is difficult to detect mutagens with lillte or no toxicity in this assay since such agents are generally weak mutagens and produce only two to three-fold increases in mutant counts. Variations of two to three-fold are often within normal fluctuations of the spontaneous counts, and the use of even higher concentrations of often difficult because of the likelihood of overloading the system with large quantities of the chemical. To resolve the mutagenicity of such a chemical, other assays to which statistical evaluations can be applied may be necessary.

Statistics:

no

Key result

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA1538, TA98, TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

Saccharomyces cerevisiae

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Conclusions:

The test compound did not demonstrate mutagenic activity in any of the assays conducted in this evaluation and was considered not mutagenic under these test conditions.

Executive summary:

The test compound was examined for mutagenic activity in a series of in vitro microbial assays employing Salmonella and Saccharomyces indicator organisms. The compound was tested directly and in the presence of liver microsomal enzyme preparations from Aroclor-induced rats.

The results of the tests conducted on the compound in the absence and in the presence of a metabolic system were all negative.

The test compound did not demonstrate mutagenic activity in any of the assays conducted in this evaluation and was considered not mutagenic under these test conditions.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Ames test (Brusick 1976) :

The test compound was examined for mutagenic activity in a series of in vitro microbial assays employing Salmonella and Saccharomyces indicator organisms. The compound was tested directly and in the presence of liver microsomal enzyme preparations from Aroclor-induced rats.

The results of the tests conducted on the compound in the absence and in the presence of a metabolic system were all negative.

The test compound did not demonstrate mutagenic activity in any of the assays conducted in this evaluation and was considered not mutagenic under these test conditions.

Justification for classification or non-classification

Based on the available data, no classification for mutagenicity is required for the registered substance according to the Regulation EC n°1272/2008.