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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:
12/1976 to 03/1977
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well-documented study report which meets basic scientific principles: pre-GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1977

Materials and methods

Principles of method if other than guideline:
Method: other: API procedure (see Reference). Similar to OECD test guideline 471.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent

Method

Target gene:
Not applicable
Species / strain
Species / strain / cell type:
other: Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538. Saccharomyces cerevisiae strain D4.
Details on mammalian cell type (if applicable):
Not applicable.
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S-9 fractions from Aroclor 1254-treated Sprague-Dawley rats
Test concentrations with justification for top dose:
0.001 to 5.0 microliters/plate.
Vehicle / solvent:
DMSO.
The compound was insoluble. An emulsion was formed upon shaking.
Controls
Untreated negative controls:
yes
Remarks:
Liver homogenate control plus solvent.
Negative solvent / vehicle controls:
yes
Remarks:
DMSO. No test chemical or activation system.
True negative controls:
not specified
Positive controls:
yes
Remarks:
Positive control info. Without activation: ethylmethanesulfonate, methylnitrosoguanidine, 2-nitrofluorene, or quinacrine mustard. With activation: 2-anthramine, 2-acetylaminofluorene, 8-aminoquinoline, or dimethylnitrosamine.
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) and in suspension

DURATION
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS:
- All concentrations were run in duplicate.
- If a positive response was noted, one or more additional runs was conducted.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
Plate incorporation.
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Concentrations were chosen such that the maximal concentration was equivalent to the 50% survival level.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
Saccharomyces cerevisiae
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
Plate incorporation
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Concentrations were chosen such that the maximal concentration was equivalent to the 50% survival level.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
without
Genotoxicity:
negative
Remarks:
Suspension: Increases were observed at the high dose in TA100, TA1537, and TA1538 but were not considered of adequate magnitude to constitute a positive response. Significant increases in TA98 were observed but not reproducible.
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Concentrations were chosen such that the maximal concentration was equivalent to the 50% survival level.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with
Genotoxicity:
negative
Remarks:
Suspension: Scattered increases were noted but not dose-dependent or reproducible. Problems were noted with initial assay, due to anomalous reductions in viable cell counts.
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Concentrations were chosen such that the maximal concentration was equivalent to the 50% survival level.
Vehicle controls validity:
valid
Untreated negative 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 with and without metabolic activations, all species/strains tested.

The in vitro reverse gene mutation study in bacteria and yeast to assess the genotoxicity of unleaded gasoline was negative. This finding does not warrant the classification of unleaded gasoline as a genotoxin under the new Regulation (EC) 1272/2008 on classification, labeling, and packaging of substances and mixtures (CLP) or under the Directive 67/518/EEC for dangerous substances and Directive 1999/45/EC for preparations.
Executive summary:

API PS-6 (unleaded gasoline) was examined for its potential to induce mutations in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 and Saccharamyces cerevisiae strain D4, in both the presence and absence of an S9 metabolic activation system. The doses tested were: 0.001, 0.01, 0.1, 1.0, and 5.0 microliters/plate. API PS-6 did not induce a statistically significant increase in the number of revertants at any of the dose levels tested with or without metabolic activation. The maximal dose chosen induced approximately 50% cytotoxicity. Under the conditions of this study, API PS-6 was not mutagenic in bacterial or yeast cells. This finding does not warrant the classification of unleaded gasoline as a genotoxic under the new Regulation (EC) 1272/2008 on classification, labeling, and packaging of substances and mixture (CLP) or under the Directive 67/518/EEC for dangerous substances and Directive 1999/45/EC for preparations.