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Toxicological information

Genetic toxicity: in vitro

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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:
February 2015 - September 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Contemporary study performed to GLP requirements. Study design included analytical verification of study concentrations per ECHA recommendation.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid stored at room temperature
Details on test material:
- Name of test material (as cited in study report): 1,3-dioxolane
- Substance type: Monoconstituent Substance
- Physical state: Colourless liquid
- Analytical purity: 99.978%
- Impurities (identity and concentrations):
- Composition of test material, percentage of components:
- Purity test date:
- Lot/batch No.: 18.12.2014, 09:45
- Expiration date of the lot/batch: 18 December 2015
- Stability under test conditions: Stable under conditions of study.
- Storage condition of test material: Stored at Controlled Room Temperature (15-25 degrees C, below 70% RH)

Method

Target gene:
S. typhimurium TA98 - hisD3052
S. typhimurium TA100 - hisG46
S. typhimurium TA1535 - hisG46
S. typhimurium TA1537 - hisC3076
E. coli WP2 uvrA - trpE
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Details on mammalian cell type (if applicable):
N/A
Additional strain / cell type characteristics:
other: S. typhimurium - uvrB deficient; E. coli - uvrA deficient
Metabolic activation:
with and without
Metabolic activation system:
mammalian (rat) liver S9 post-mitochondrial fraction
Test concentrations with justification for top dose:
0, 78.125, 156.25, 312.5, 625, 1250, 2500, 5000 microgram/plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: Selection of water was based on solubility testing.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
methylmethanesulfonate
other: 4-nitro-1,2-phenylene-diamine (NPD); 2-aminoanthracene (2AA)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) used for Initial Mutation Test; preincubation used for Confirmatory Mutation Test

DURATION
- Preincubation period: For Confirmatory Mutation Test, preincubation period was 20 min.
- Exposure duration: 48 hours post exposure.

SELECTION AGENT (mutation assays): N/A for reverse mutation assays.

NUMBER OF REPLICATIONS: Study included a Preliminary Concentration Range Finding Test, an Initial Mutation Test, and a Confirmatory Mutation Test. Each concentration/condition (+/- S9) was evaluated in triplicate.



DETERMINATION OF CYTOTOXICITY
- Method: Preliminary Concentration Range Finding Test performed to ensure lack of cytotoxicity, appropriateness of dose levels.
Evaluation criteria:
Criteria for a Positive Response:

A test item was considered mutagenic if:
- a dose–related increase in the number of revertants occurred and/or;
- a reproducible biologically relevant positive response for at least one of the dose groups occurred in at least one strain with or without metabolic activation.

An increase was considered biologically relevant if:

- the number of reversions was more than two times higher than the reversion rate of the negative (solvent) control in Salmonella typhimurium TA98, TA100 and Escherichia coli WP2 uvrA bacterial strains;
- the number of reversions was more than three times higher than the reversion rate of the negative (solvent) control in Salmonella typhimurium TA1535 and TA1537 bacterial strains.

Criteria for a Negative Response:

The test item was considered to have shown no mutagenic activity in this study if it produced neither a dose-related increase in the number of revertants nor a reproducible biologically relevant positive response at any of the dose groups, with or without metabolic activation.
Statistics:
The mean number of revertants per plate, the standard deviation and the mutation factor* values were calculated for each concentration level of the test item and for the controls using Microsoft Excel software.

Results and discussion

Test results
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
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test item 1,3-Dioxolane had no mutagenic activity in the examined bacterial strains under the test conditions of this study.
Executive summary:

The test item 1,3-Dioxolane was tested for potential mutagenic activity using the Bacterial Reverse Mutation Assay.

The experiments were carried out using histidine-requiring auxotroph strains of Salmonella typhimurium (Salmonella typhimurium TA98, TA100, TA1535 and TA1537), and the tryptophan-requiring auxotroph strain of Escherichia coli (Escherichia coli WP2 uvrA) in the presence and absence of a metabolic activation system, which was a cofactor-supplemented post-mitochondrial S9 fraction prepared from the livers of phenobarbital/β-naphthoflavone-induced rats.

The study included a Preliminary Compatibility Test, a Preliminary Concentration Range Finding Test, an Initial Mutation Test and a Confirmatory Mutation Test. In the Preliminary Concentration Range Finding Test as well as in the Initial Mutation test, the plate incorporation method was used. In the Confirmatory Mutation Test, the pre-incubation method was used. The reported data of this mutagenicity assay show that under the experimental conditions applied the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

In conclusion, the test item 1,3-Dioxolane had no mutagenic activity in the examined bacterial strains under the test conditions of this study.