Distillates (petroleum), light distillate hydrotreating process, low-boiling

Genetic toxicity: in vitro

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

Test material information

Constituent 1

Reference substance name:

unleaded gasoline

IUPAC Name:

unleaded gasoline

Constituent 2

Reference substance name:

PS-6

IUPAC Name:

PS-6

Constituent 3

Reference substance name:

Gasoline

EC Number:

289-220-8

EC Name:

Gasoline

Cas Number:

86290-81-5

IUPAC Name:

Gasoline

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 results

Test results 1

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

Test results 2

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

Test results 3

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

Test results 4

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.