Diethyl ether

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented, according to accepted guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Diethyl ether

EC Number:

200-467-2

EC Name:

Diethyl ether

Cas Number:

60-29-7

Molecular formula:

C4H10O

IUPAC Name:

diethyl ether

Details on test material:

- Name of test material (as cited in study report): Diethyl ether
- Physical state: Clear liquid
- Lot/batch No.: 09023198
- Expiration date of the lot/batch: 09/2010
- Storage condition of test material: At room temperature

Method

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital and β-naphthoflavone-induced rat liver microsomal fraction (S9)

Test concentrations with justification for top dose:

Toxicity experiment (experiment I): 0.156, 0.312, 0.625, 1.25, 2.5, or 5 µL/mL (± S9)
Toxicity experiment (experiment II): 0.5, 1, 2, 3, 4, or 5 µL/mL (- S9)

Mouse lymphoma assay (Experiment I): 0.039, 0.078, 0.156, 0.312, 0.625, 1.25, 2.5, or 5 µL/mL (± S9)
Mouse lymphoma assay (Experiment II): 0.1, 0.2, 0.5, 1, 2, 3, 4, or 5 µL/mL (± S9)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: RPMI + 3% horse serum (for short-term exposure) or RPMI + 7.5% horse serum (for long-term exposure)

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

- RPMI + horse serum (3% of 7.5%)

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Ethylmethanesulfonate and methylmethanesulfonate (- S9) and benzo[a]pyrene (+ S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 hours (for short-term exposure; ± S9) and 24 hours (for long-term exposure; - S9) (temperature not reported)
- Expression time (cells in growth medium): 72 hours (for short-term exposure) and 48 hours (for long term exposure) at 37 ºC

NUMBER OF REPLICATIONS: Duplicate plates/dose; independent repeat experiment was performed.

Evaluation criteria:

Criteria for determining a positive result:

- Clear and dose-related increase in the mutation frequency

- Biologically relevant response (at least a 2-fold increase of mutant frequencies related to the respective negative control values and higher than the historical range of negative controls) for at least one of the dose groups.

- Combined with a positive effect in the mutant frequency, an increased occurrence of small colonies (slow growth colonies) indicated by a low large/small colonies ratio (1.5 times the ratio of clastogenic controls MMS and/or B[a]P) is an indication for potential clastogenic effects and/or chromosomal aberrations.

A test item is considered to be negative if there is no biologically relevant increase in the induction of mutant cells above concurrent control levels, at any dose level.

Statistics:

Not performed but not considered necessary.

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

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 with metabolic activation
negative without metabolic activation