Ethyl 2,4-dimethyl-1,3-dioxolane-2-acetate

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:

13 March 2002 - 28 March 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

dd 11 March 2002

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

Ethyl 2,4-dimethyl-1,3-dioxolane-2-acetate

EC Number:

228-536-2

EC Name:

Ethyl 2,4-dimethyl-1,3-dioxolane-2-acetate

Cas Number:

6290-17-1

Molecular formula:

C9H16O4

IUPAC Name:

ethyl 2-(2,4-dimethyl-1,3-dioxolan-2-yl)acetate

Test material form:

liquid

Method

Target gene:

Histidine gene

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97a, TA98, TA100, TA102 and TA1535

Metabolic activation:

with and without

Metabolic activation system:

S9 from livers of male Wistar rats, induced with phenobarbital and ß-naphtoflavone

Test concentrations with justification for top dose:

0.05, 0.16, 0.5, 1.6 and 5 mg/plate
The top dose was selected according to OECD guideline 471.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water (aqua bidest)
- Justification for choice of solvent/vehicle: according to OECD guideline 471.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

cumene hydroperoxide

other: ICR 191, 4-nitro-o-phenylendiamine, nitrofurantoine, 2-aminoanthracen, danthron

Remarks:

See table 1 for details on positive control substances

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar
- Medium used: 25 mL Vogel-Bonner minimal medium; for strain TA97a with 0.4% D+ glucose, for all other strains with 2% D+ glucose

DURATION
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3 per concentration and control (titer control replicates were prepared 2-fold).

DETERMINATION OF CYTOTOXICITY
- Method: plates were inspected for present and reduced background lawn; colonies were counted, if no reduced background lawn was observed, with a colony counter (BZG 30; WTW)
- Any supplementary information relevant to cytotoxicity:

- OTHER: The test item is to be interpretated mutagenic if there is a concentration effect relationship and the induction rate is ≥ 2.

Evaluation criteria:

Since no validity criteria are described in OECD and EC guidelines, the following validity criteria from the literature and DIN guideline were used:
- The following genotypes of the tested strains had to be confirmed:
- Histidine auxotrophy
- Ampicillin resistance
- Tetracycline resistance (only TA 102)
- UV-sensitivity (except TA 102)
- Growth inhibition with crystal violet (rfa-mutation)
- Titer of the overnight culture had to be ≥ 1 x 10^8 cells/mL.
- Spontaneous revertants/plate (negative controls) had to be within the following ranges:
- TA 97 a ± S9 : 150 - 450
- TA 98 ± S9 : 15- 50
- TA 100 ± S9 : 60 - 200
- TA 102 ± S9 : 300 - 600
-TA 1535 ± S9 : 5- 30
- The induction rates of the positve controls had to be ≥ 2.

The validity criteria were met.

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

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 98

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

Test results 3

Key result

Species / strain:

S. typhimurium TA 100

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

Test results 4

Key result

Species / strain:

S. typhimurium TA 102

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

Test results 5

Key result

Species / strain:

S. typhimurium, other: TA97a

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

Applicant's summary and conclusion

Conclusions:

The substance is not mutagenic in the Salmonella typhimurium reverse mutation assay performed according to OECD 471 guideline and GLP principles.

Executive summary:

The mutagenic activity of the substance was evaluated in accordance with OECD 471 guideline and according to GLP principles. The test was performed in two independent experiments in the absence and presence of S9 -mix. Adequate negative and positive controls were included.

The maximum dose level of the test item in both experiments was selected as the maximum recommended dose level of 5000 μg/plate. There was no visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation (S9-mix) in both experiments. The substance did not induce a significant dose related increase in the number of revertant (His+) colonies in each of the five S.typhimurium tester strains (TA97a, TA1535, TA102, TA98 and TA100) both in the absence and presence of S9-metabolic activation. These results were confirmed in the independently repeated experiment. Based on the results of this study it is concluded that the substance is not mutagenic in the Salmonella typhimurium reverse mutation assay.