Fatty acids, C18-unsatd., dimers, ethoxylated

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:

2012-07-11 until 2012-07-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline and GLP compliant study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

, adopted 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OTS 798.5100 (Escherichia coli WP2 and WP2 UVRA Reverse Mutation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C18-unsatd., dimers, ethoxylated

EC Number:

614-587-1

Cas Number:

68551-92-8

Molecular formula:

Unspecified

IUPAC Name:

Fatty acids, C18-unsatd., dimers, ethoxylated

Test material form:

other: liquid

Method

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 mix of Phenobarbital/ß-naphtoflavone induced rats.

Test concentrations with justification for top dose:

0; 33; 100; 333; 1 000; 2 750 and 5 500 μg/plate

Vehicle / solvent:

The test substance was dissolved in DMSO.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other:

Remarks:

With S9: 2-aminoanthracene (all strains), Without S9: N-methyl-N'-nitro-N-nitrosoguanidine (TA 1535, TA 100); 4-nitro-o-phenylenediamine (TA98); 9-aminoacridine (TA 1537); 4-nitroquinoline-N-oxide (E. coli WP2 uvrA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: plate incorporation (experiment I) preincubation test (experiment II)
DURATION: Plate incorporation: 48 hours at 37°C, pre-incubation for 20 min at 37°C followed by a 48 - 72 h incubation of the prepared plates
NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY:
• decrease in the number of revertants
• clearing or diminution of the background lawn (= reduced his- or trp- background growth)
• reduction in the titer (the titer is determined only in the experimental parts with S9 mix both for the
negative controls (vehicle only) and for the two highest doses in all experiments).

Evaluation criteria:

Acceptance criteria
Generally, the experiment is considered valid if the following criteria are met:
- The number of revertant colonies in the negative controls was within the range of the
historical negative control data for each tester strain.
- The sterility controls revealed no indication of bacterial contamination.
- The positive control substances both with and without S9 mix induced a distinct increase in
the number of revertant colonies within the range of the historical positive control data or
above.
- Fresh bacterial culture containing approximately 109 cells per mL were used. For approval
the titer of viable bacteria was ≥ 108 colonies per mL.

Assessment criteria
The test substance is considered positive in this assay if the following criteria are met:
- A dose-related and reproducible increase in the number of revertant colonies, i.e. about
doubling of the spontaneous mutation rate in at least one tester strain either without
S9 mix or after adding a metabolizing system.
A test substance is generally considered non-mutagenic in this test if:
- The number of revertants for all tester strains were within the historical negative control
range under all experimental conditions in at least two experiments carried out
independently of each other.

Statistics:

not applicable

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5500 µg/plate in the standard plate test with S9; occasionally in the pre-incubation assay at test concentrations from 2750 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

A weak bacteriotoxic effect (slight decrease in the number of his+ revertants, slight reduction in the titer) was observed in the standard plate test only with metabolic activation at 5500 μg/plate using the salmonella strains.
In the preincubation assay bacteriotoxicity (slight decrease in the number of his+ revertants, slight reduction in the titer) was occasionally observed depending on the strain and test conditions from 2750 μg/plate onward using the salmonella strains.
Test substance precipitation was found from 2750 μg/plate onward with and without S9 mix.

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

Conclusively, the test item is not a mutagenic test substance in the bacterial reverse mutation test in the absence and the presence of metabolic activation.