2,3-Epoxypropyl neodecanoate, oligomeric reaction products with cyclohexane-1,2-dicarboxylic anhydride and propylidenetrimethanol

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:

weight of evidence

Study period:

06 June 2017 to 11 August 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,3-Epoxypropyl neodecanoate, oligomeric reaction products with cyclohexane-1,2-dicarboxylic anhydride and propylidenetrimethanol

EC Number:

500-334-1

EC Name:

2,3-Epoxypropyl neodecanoate, oligomeric reaction products with cyclohexane-1,2-dicarboxylic anhydride and propylidenetrimethanol

Cas Number:

154565-28-3

Molecular formula:

A molecular formaula is not available for this UVCB

IUPAC Name:

2,3-Epoxypropyl neodecanoate, oligomeric reaction products with cyclohexane-1,2-dicarboxylic anhydride and propylidenetrimethanol

Test material form:

solid

Remarks:

Clear yellow resin

Specific details on test material used for the study:

Lot number: CVR83297
Purity: 97.2%
Molecular weight: 1281.67 g/mol
Appearance: Clear yellow-orange resin
Storage: Room temperature; protected from light
Expiry date: 27 December 2018

Method

Target gene:

The Salmonella strains contain mutations in the histidine operon, thereby imposing a requirement for histidine in the growth medium. These strains contain the deep rough (rfa) mutation, which deletes the polysaccharide side chain from the lipopolysaccharides of the bacterial cell surface. This increases cell permeability of larger substances. The other mutation is a deletion of the uvrB gene, which codes for a protein of the DNA nucleotide excision repair system, resulting in an increased sensitivity in detecting many mutagens. This deletion also includes the nitrate reductase (chi) and biotin (bio) genes (bacteria require biotin for growth). Tester strains TA98 and TA100 contain the R-factor plasmid, pKM101. These strains are reverted by a number of mutagens that are detected weakly or not at all with the non-R-factor parent strains. pKM101 increases chemical and spontaneous mutagenesis by enhancing an error-prone DNA repair system, which is normally present in these organisms. The tester strain Escherichia coli WP2 uvrA carries the defect in one of the genes for tryptophan biosynthesis. Tryptophan-independent mutants (revertants) can arise either by a base change at the site of the original alteration or by a base change elsewhere in the chromosome so that the original defect is suppressed. This second possibility can occur in several different ways so that the system seems capable of detecting all types of mutagens, which substitute one base for another. Additionally, the strain is deficient in the DNA nucleotide excision repair system.

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9

Test concentrations with justification for top dose:

In the initial toxicity-mutation assay, the dose levels tested were 1.50, 5.00, 15.0, 50.0, 150, 500, 1500 and 5000 µg per plate.Based upon these results, the maximum dose tested in the confirmatory mutagenicity assay was 5000 µg per plate.In the confirmatory mutagenicity assay, the dose levels tested were 15.0, 50.0, 150, 500, 1500 and 5000 µg per plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Dimethyl sulfoxide (DMSO) for the test substance; all positive controls were diluted in DMSO except for sodium azide, which was diluted in sterile water - Justification for choice of solvent/vehicle: DMSO was the vehicle of choice based on the solubility of the test substance and compatibility with the target cells.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (pre-incubation: incubated with shaking for 60±2 minutes at 37±2°C)
DURATION: Exposure duration: 48 to 72 hours
NUMBER OF REPLICATIONS: 2 in the initial-toxicity mutation assay; 3 in the confirmatory mutagenicity assay
NUMBER OF CELLS EVALUATED: >/= 0.3 x 10^8 cells/plate
DETERMINATION OF CYTOTOXICITY- Method: other: Counting of revertant colony numbers and evaluation of the condition of the bacterial background lawn.

Evaluation criteria:

For the test substance to be evaluated positive, it must cause a dose-related increase in the mean revertants per plate of at least one tester strain over a minimum of two increasing concentrations. Strains TA1535 and TA1537Data sets were judged positive if the increase in mean revertants at the peak of the dose response was equal to or greater than 3.0-times the mean vehicle control value and above the corresponding acceptable vehicle control range. Strains TA98, TA100 and WP2 uvrA were judged positive if the increase in mean revertants at the peak of the dose response was equal to or greater than 2.0-times the mean vehicle control value and above the corresponding acceptable vehicle control range.An equivocal response is a biologically relevant increase in a revertant count that partially meets the criteria for evaluation as positive. This could be a dose-responsive increase that does not achieve the respective threshold cited above or a non-dose responsive increase that is equal to or greater than the respective threshold cited. A response was evaluated as negative if it was neither positive nor equivocal.

Statistics:

According to the test guidelines, the biological relevance of the results is the criterion for the interpretation of the results, and a statistical evaluation of the results is not regarded as necessary.

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
Precipitation: Precipitate was observed beginning at 1500 or at 5000 µg per plate with all conditions in intial-toxicity mutation assay and confirmatory mutagenicity assay.

Applicant's summary and conclusion

Conclusions:

All criteria for a valid study were met as described in the protocol. The results of the study indicate that, under the conditions of this study, Epoxypropyl neodecanoate, oligmeric reaction products with cyclohexane-1,2-dicarboxylic anhydride and propylidenetrimethanol did not cause a positive mutagenic response with any of the tester strains in either the presence or absence of Aroclor induced rat liver S9.

Executive summary:

The test substance [Epoxypropyl neodecanoate, oligmeric reaction products with cyclohexane-1,2-dicarboxylic anhydride and propylidenetrimethanol] was tested in an Ames test to evaluate its mutagenic potential by measuring its ability to induce reverse mutations Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and in Escherichia coli strain WP2uvrA, in the presence and absence of an exogenous metabolic activation system (induced rat liver S9 fraction).  Dimethyl sulphoxide (DMSO) was used as the vehicle. In an initial assay, concentrations of 1.50, 5.00, 15.0, 50.0, 150, 500, 1500 and 5000 ¿g/plate were tested. No toxicity was observed. Precipitation of the test material was observed at concentrations of 1500 and 5000 ¿g/plate. No positive responses were observed with any of the tester strains in either the presence or absence of S9 activation. In a confirmatory mutagenicity assay, the concentrations tested were 15.0, 50.0, 150, 500, 1500 and 5000 ¿g/plate. No toxicity was observed. Precipitate was observed beginning at 1500 and 5000 ¿g/plate. No positive responses were observed with any of the tester strains in either the presence or absence of S9 activation. The results of this study indicate that the test material [Epoxypropyl neodecanoate, oligmeric reaction products with cyclohexane-1,2-dicarboxylic anhydride and propylidenetrimethanol] is not mutagenic under the conditions of the assay.