Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
US EPA, Method: HG-Gene MUta-S. typhimurium; the Salmonella typhimurium reverse mutation assay, 1984
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Substance: FR -522 Purified (2,2-bis(bromomethyl)-1,3-propanediol), Purity: 99.5%,

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 TA 98 and TA 100 possess a plasmid (pKM101) which introduce an error prone repair process resulting increased sensitivity to mutagens
Metabolic activation:
with and without
Metabolic activation system:
liver preparations from Aroclor 1254 induced rats and uninduced syrian hamsters.
Test concentrations with justification for top dose:
Preliminary toxicity test: Solvent, 5, 50, 500, 5000 microgram/plate in the presence and absence of liver S-9 mix/Hamster S-9 mix
Mutation tests: Solvent, 0, 50, 150, 500, 1500, 5000 microgram/plate in the presence and absence of liver S-9 mix/Hamster S-9 mix
Vehicle / solvent:
Solvent: Dimethylsulphoxide
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
Dimethylsulphoxide (DMSO)
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
In the absence of S-9 mix
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
In the absence of S-9 mix
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
In the absence of S-9 mix
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
In the presence of S-9 mix
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
congo red
Remarks:
In the presence of S-9 mix
Details on test system and experimental conditions:
Test Type: Reverse Mutation Assay
System of testing: Bacterial
Species/Strain: Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537
Metabolic activation: S9-mix,Rat liver cells, Aroclor induced; S9-mix,Hamster liver cells, uninduced
Test Design
Number of replicates: 3
Positive Controls: N-ethyl -N’-nitroso-N-nitrosoguanidine (TA100, TA1535, -S9)
9-aminoacridine (TA1537, -S9)
2-nitrofluorene (TA98, -S9); 2-aminoanthracene (TA98, TA100, TA 1535, TA1537, +S9)
Congo red (TA98, +S9)
Negative Control: Solvent vehicle
Solvent: Dimethylsulfoxide
(see attached experimental procedure)
Evaluation criteria:
See attached document (assessment of results)
Statistics:
see attached document (statistical results AMES 001)

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Remarks:
up to 5000 microgram/plate
Vehicle controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
positive
Remarks:
dose related increase only in the presence of hamster S-9 mix and were largest in the presence of the 30% mix
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Remarks:
up to 5000 microgram/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
positive
Remarks:
using 30% Hamster S-9 mix
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Result: Positive
Cytotoxic concentration -With metabolic activation: > 5000 μg/plate -Without metabolic activation:> 5000 μg/plate
Genotoxic effects:
With metabolic activation: Large dose-related increases in revertant colony numbers were observed with strain TA1535.
These increases were observed only in the presence of hamster S-9 mix and were largest in the presence of the 30% mix.
A smaller increase was also observed with strain TA100 in the presence of 30% hamster S-9 mix.
Without metabolic activation:None
Remarks on result:
other:
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

see attached table (statistical results)

Remark: S-9 is usually only prepared from a rat previously treated with a compound (Aroclor) known to induce a high level of enzymic activity. However in this study S-9 mix using liver fraction from uninduced Syrian hamsters was also used as it was suspected that the test substance may be metabolised to a mutagen by hamster S-9 but not by rat.

Applicant's summary and conclusion

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

It is concluded that, when tested in dimethylsulfoxide, the test substance shows no evidence of mutagenic activity in the absence or presence of rat S-9 mix. The test substance shows clear evidence of mutagenic activity with strains TA1535 and TA100 in the presence of hamster S-9 mix.

Executive summary:

It is concluded that, when tested in dimethylsulfoxide, the test substance shows no evidence of mutagenic activity in the absence or presence of rat S-9 mix. The test substance shows clear evidence of mutagenic activity with strains TA1535 and TA100 in the presence of hamster S-9 mix.