Registration Dossier

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: guideline study under GLP consitions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: amber viscous liquid
Details on test material:
Content: 100 & Additin E459
Main Compound: Triethyleneglycol dimaleate (86 %)
Byproduct: triethyleneglycol monomaleate (9 %)

Method

Target gene:
gene mutation by base-pair changes or frame-shifts in the genome
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Additional strain characteristics:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
The S9 fraction was prepared form the livers of Aroclor 12254-induced male Sprague-Dawley rats.The S9-mix was freshly prepared, kept on ice and used only on the same day.
Test concentrations with justification for top dose:
0 (solvent control), 50, 160, 500, 1600 5000 µg/plate
Vehicle:
demineralized water
Controls
Negative controls:
yes
Solvent controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
2-nitrofluorene
sodium azide
other: 4-Nitro-o-phenylene diamine, Mitomycin C, Cumene hydroperoxide, Anthracene-2-amine
Details on test system and conditions:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102. pre-incubation methodology and plate incorporation methodology, 2 independent trials with S9-mix with each methodoloy snd 2 independent trials without S9-mix with each methodoloy
Evaluation criteria:
A reproducible and dose-related increase in mutant counts of at least one strain is considered to be positive result.
For TA 1535, TA100, TA1537 and TA98 this increase should be about twice of the solvent controls. For TA102 an increase of about 100 mutants should be reached. Otherwise, the result is evaluated as negative.
However, these criteria may be overruled by good scientific judgement.
In questionable results, investgations should continue, possibly with modifications, until a final evaluation is possible.
Statistics:
no data

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Additional information on results:
None of the 5 strains used showed a dose-related and biologically relevant increase in mutant counts over those of the solvent controls in the plate incorporation test. This applied both to the tests with and without S9-mix and was confirmed by the results of the pe-incubation trials.No bacteriotoxic effects or precipitations of the substance were observed.
The positive controls increased mutant counts to well over the solvent controls, and thus demonstrated the sysem's sensitivity and the activity of the S9-mix

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative
Executive summary:

Additin E459 was investigated for point mutagenic effects according to OECD TG 471 and GLP using S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence and in the absence of a metabolic activation systen. The test was performed using the the plate incorporation and the pre-incubation methodology with doses up to and including 5000 µg/plate.

None of the 5 strains used showed a dose-related and biologically relevant increase in mutant counts over those of the solvent controls in the plate incorporation test. This applied both to the tests with and without S9-mix and was confirmed by the results of the pe-incubation trials.No bacteriotoxic effects or precipitations of the substance were observed. The positive controls increased mutant counts to well over the solvent controls, and thus demonstrated the sysem's sensitivity and the activity of the S9-mix.

Therefore, Additin E 459 was considered to be non-mutagenic in the Ames test under the conditions reported.