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Toxicological information

Genetic toxicity: in vitro

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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:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
mammalian cell gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
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