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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: clear colorless liquid
Details on test material:
content/purity: 98.7 %

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Additional strain / cell type 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 from the livers of Aroclor 1254-induced male Sprague Dawley rats
Test concentrations with justification for top dose:
-----plate incorporation methodology,
S. typh TA1535, TA100, TA1537, TA98, TA102 with and without S9-mix
0, 50, 160, 500, 1600, 5000 µg/plate
--additionally S. typh TA100, TA1537 with and without S9-mix
0, 1.6 , 5, 16 µg/plate
-----preincubation methodology
S. typh TA 1535 with and without S9-mix
0, 1.6, 5, 16, 50, 160, 500, 1600 µg/plate
S. typh TA100, TA1537 with and without S9-mix
0, 1.6, 5, 16, 50, 160, 500 µg/plate
S typh TA98 with and without S9-mix
0, 16, 50, 160, 500, 1600 µg/plate
S typh TA102 with and without S9-mix
0, 50, 160, 500, 1600. 5000 µg/plate
-- additionally S typh TA1535, TA98 without S9-mix
0, 1.6, 5 µg/plate
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
2-nitrofluorene
sodium azide
cumene hydroperoxide
mitomycin C
other: 4-Nitro-1,2-phenylenediamine, 2-aminoanthracene
Details on test system and experimental conditions:
According to OECD TG 471 ( plate incorporation methodology and pre-incubation methodology
Evaluation criteria:
A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result. For TA1535, TA100, TA1537 and TA98 this increase should be about twice that of 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 judgment.
In case of questionable results, investigations 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 / choice of top concentrations:
other: plate incorporation test strain-specific at 160 µg/plate and above bacteriotoxic, precipitation at 5000 µg/plate: pre-incubation: stain specific bacteriotoxic at 16 µg/plate and above; precipitation at 5000 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested

Applicant's summary and conclusion

Conclusions:
Interpretation of results: negative
Executive summary:

The principal aim of this study was the investigation of the test item for point mutagenic effects using a plate incorporation test and a repeat test with preincubation modification. For this purpose, the test item, dissolved in DMSO, was administered in initially doses of up to and including 5000 μg per plate without and with S9 mix on the five Salmonella typhimurium LT2 mutant strains TA1535, TA100, TA1537, TA98 and TA102, according to the OECD guideline 471 under GLP conditions. In the plate incorporation test strain-specific bacteriotoxic were seen at 160 μg per plate.and above and in the pre-incubation test at 16 µg/plate and above, In both methodologies substance precipitation occurred at 5000 μg per plate.

Under the experimental conditions reported (direct plate incorporation procedure and preincubation modification) the test item did not induce gene mutations by base-pair changes or frame-shifts in the genome of the S. typhimurium strains used, when tested up to a maximum recommended dose of 5000 μg/plate in the absence and presence of S9 mix.