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Genetic toxicity: in vitro

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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:
1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and appropriate guidelines
Cross-reference
Reason / purpose for cross-reference:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Qualifier:
according to guideline
Guideline:
other: US EPA Method: HG-GENE Muta-S. typhimurium: The salmonella typhimurium revers mutation assay 1984
Principles of method if other than guideline:
not relevant
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2-dimethylpropan-1-ol, tribromo derivative
EC Number:
253-057-0
EC Name:
2,2-dimethylpropan-1-ol, tribromo derivative
Cas Number:
36483-57-5
Molecular formula:
C5H9Br3O
IUPAC Name:
3-bromo-2,2-bis(bromomethyl)propan-1-ol
Details on test material:
Identity: FR-513
Appearance: Off-white, flaky solid
Storage: Room temperature in the dark
Purity: 98%
Stable for at least two years

Method

Target gene:
histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
not relevant
Additional strain / cell type characteristics:
other: Strains TA98 and TA1537 were capable of detecting frameshift mutagens, strains TA100 and TA1535 are capable of detecting base-pair substitution mutagens
Metabolic activation:
with and without
Metabolic activation system:
liver preparation from Aroclor 1254-induced rats (uninduced syrian hamsters) rats and unindiced
Test concentrations with justification for top dose:
Concentration range in Preliminary toxicity determination (with/ without metabolic activation): 0, 5, 50, 500, 5000 µg/plate
Concentration range in the main test (with/without metabolic activation): solvent, 0, 15, 50, 150, 500, 1500 µg/plate
Vehicle / solvent:
Solvent: DMSO
Controls
Untreated negative controls:
yes
Remarks:
DMSO
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
yes
Remarks:
no test substance
Positive controls:
yes
Remarks:
N-Ethyl-N-nitro-N-nitrosoguanidine, 9-Amonoacridine, 2 Nitrofluorene, 2-Aminoantracene, Congo red
Details on test system and experimental conditions:
see attached document on experimental procedure
Evaluation criteria:
See attached document on assessment of results
Statistics:
no

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
5-500 µg/plate
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
5000 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
without
Genotoxicity:
negative
Remarks:
0-500 µg/plate
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1500µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with
Genotoxicity:
ambiguous
Remarks:
positive only in the presence of hamster S-9 mix (15-500 µg/plate)
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1500 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with
Genotoxicity:
ambiguous
Remarks:
positive only in the presence of hamster S-9 mix (15-500 µg/plate)
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1500 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium, other: TA 98, TA 1537
Metabolic activation:
with
Genotoxicity:
negative
Remarks:
15-500 µg/plate
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1500 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Preliminary test: FR-513 was toxic towards all tester strains at 5000 µg/plate. Therefore, 1500 µg/plate was chosen at the top dose level in the mutation test.
Main tests: Following treatment with FR-513, large, dose-related increases in revertant colony numbers were observed in both mutation tests with
strains TA 1535 and TA 100. These increases were observed only in the presence of hamster S-9 mix and were largest in the present of the 30% mix.
The concurrent positive control compounds demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations.
Remarks on result:
other: other: preliminary toxicity test
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

See attached document on tables

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
ambiguous with metabolic activation

It concluded that when tested in dimethyl sulphoxide, FR-513 shows no evidence of mutagenic activity in the absence or presence of rat S-9 mix.
FR-513 shows clear evidence of mutagenic activity between 500 and 15 µg/plate with strains TA 1535 and TA 100 in the presence of hamster S-9 mix.
Executive summary:

In vitro assesment of mutagenic potential of FR-513, histidine dependant auxtrophic mutants of S. typhimurium (Strains TA 1535, TA 1537, TA 98 and TA 100) was conducted according to the appropriate OECD guidelines such as OECD 471. Two independant mutation tests were conducted in the presence and absence of liver preparations from rats and hamsters.

It was concluded that when tested in dimethyl sulphoxide, FR-513 shows no evidence of mutagenic activity in the absence or presence of rat S-9 mix. FR-513 shows clear evidence of mutagenic activity between 500 and 15 µg/plate with strains TA 1535 and TA 100 in the presence of hamster S-9 mix.