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
Study period:
2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in 2014 study at recognised contract research organisation.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Batch SC00008199
Purity 99.5%
Expiry date 19 August 2016

Method

Target gene:
Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2uvrA.
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
10% liver S9 in standard co-factors
Test concentrations with justification for top dose:
Experiment 1: 1.5, 5, 15, 50, 150, 500, 1500 and 5000 μg/plate) with and without metabolic activation.
Experiment 2:
Salmonella strains TA1535 and TA98 (absence of S9-mix) and E.coli strain WP2uvrA (absence and presence of S9-mix): 0.5, 1.5, 5, 15, 50, 150, 500 μg/plate.
Salmonella strains TA100 (absence and presence of S9-mix), TA1535 (presence of S9-mix) and TA1537 (absence of S9-mix): 5, 15, 50, 150, 500, 1500, 5000 μg/plate.
Salmonella strains TA98 and TA1537 (presence of S9-mix): 15, 50, 150, 500, 1500, 5000 μg/plate.
Vehicle / solvent:
Dimethyl sulphoxide was selected as the vehicle.
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
2 μg/plate for WP2uvrA without S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
3 μg/plate for TA100 without S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
5 μg/plate for TA1535 without S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
80 μg/plate for TA1537 without S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
0.2 μg/plate for TA98 without S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene (2AA)
Remarks:
1 μg/plate for TA100 with S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene (2AA)
Remarks:
2 μg/plate for TA1535 and TA1537 with S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
dimethyl sulphoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene (2AA)
Remarks:
10 μg/plate for WP2uvrA with S9-mix
Untreated negative controls:
yes
Remarks:
Vehicle, negative (untreated)
Negative solvent / vehicle controls:
yes
Remarks:
Vehicle, negative (untreated)
True negative controls:
no
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
Remarks:
5 μg/plate for TA98 with S9-mix

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
see additional information on results
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'.

Any other information on results incl. tables

The first experiment was selected as the maximum recommended dose level of 5000 μg/plate. In the first mutation test (plate incorporation method), the test item caused a visible reduction in the growth of the bacterial background lawns of all of the Salmonella tester strains initially from 1500 μg/plate in both the absence and presence of S9-mix. No toxicity was noted to Escherichia coli strain WP2uvrA either in the absence or presence S9-mix.

In the second mutation test (pre-incubation method), the test item induced a stronger toxic response with weakened bacterial background lawns noted in the absence of S9-mix from 150 μg/plate (TA1535 and TA98) and 500 μg/plate (TA100, TA1537 and WP2uvrA). In the presence S9-mix weakened bacterial background lawns were noted from 500 μg/plate (TA100, TA1535 and WP2uvrA) and 1500 μg/plate (TA98 andTA1537).

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Cumin Nitrile was considered to be non-mutagenic under the conditions of this test.