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Toxicological information

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:
1990-08-30 to 1990-09-03
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study with acceptable restrictions: TA 102 or E.coli WP2 were not tested (not required by applied version of guideline).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
Cited as Directive 84/449/EEC, B.14
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Cyclooctene-cis of Hüls AG, purity 96.81 %, produced 01 Mar 1990, ID 3641/81173

Method

Target gene:
mutated gene loci responsible for histidine auxotrophy
Species / strain
Species / strain / cell type:
other: Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98, TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat S9 liver
Test concentrations with justification for top dose:
8; 40; 200; 1000; 5000 µg/plate
Vehicle / solvent:
acetone
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
acetone
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
sodium azide for TA 100 and 1535, aminoacridine for TA 1537

Migrated to IUCLID6: TA 98 and 1538
Details on test system and experimental conditions:
Ames test
SYSTEM OF TESTING
- Metabolic activation system:   Aroclor 1254 induced rat S9 liver, male Bor: WISW (SPF/Cpb) rats
ADMINISTRATION: 
- Dosing:    
main test: 8/40/200/1000/5000 µg/plate (+/- metabolic activation)   
pre-incubation test: 8/40/200/1000/5000 µg/plate (+/- metabolic  activation)
- Number of replicates: 3
- Application: solvent acetone (CAS No. 67-64-1)   main test 100 g/l, pre-incubation test 200 g/l
- Positive and negative control groups and treatment:    
positive, TA 98 and TA 1538: nitrofluorene   
positive, TA 100 and TA 1535: sodium azide   
positive, TA 1537: aminoacridine   
negative: solvent + untreated controls   activity of metabolic system: aminoanthracene / TA 100
- Pre-incubation: 30 minutes at 30 °C   incubation 96 hours at 37 °C (approximately)
Evaluation criteria:
CRITERIA FOR EVALUATING RESULTS:    mutagenic effects (i.e  ratio of revertant rates treated/control >= 2)  at <= 5000 µg/plate with generally 
positive dose-response relationship in  any strain
Statistics:
usual statistical methods (standard deviation; means; factors) were calculated through a computer program by Messrs BIOSYS.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98, TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: 5000 µg/plate in TA 1535 and TA 1537, only without S9 mix
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
GENOTOXIC EFFECTS: 
- With metabolic activation: None
- Without metabolic activation: None   
The positive controls were functional.
PRECIPITATION CONCENTRATION: 
5000 µg/plate in pre-incubation test
CYTOTOXIC CONCENTRATION (including effects on background lawn):    
TA 98: none (+/- S9)   
TA 100: none (+/- S9)   
TA 1535: 5000 µg/plate (- S9)   
TA 1537: 5000 µg/plate (- S9)  
TA 1538: none (+/- S9)
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Cis-cyclooctene was not mutagenic when tested on S. typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538, with or without S9-mix activation.
Executive summary:

In a reverse gene mutation assay in bacteria, strains TA98, TA100, TA1535, TA1537 and TA1538 of Salmonella typhimurium were exposed to Cis-cyclooctene using acetone as a vehicle at concentrations of 8, 40, 200, 100 and 5000 µg/plate, with and without S-9 activation in a preincubation assay according to EEC guidance Dir. 84/449/EEC, B.14.

The test article was tested at six dose levels along with appropriate vehicle and positive controls on the tester strains mentioned above in the presence and absence of S9-mix. All dose levels, vehicle and positive controls were plated in triplicate.

Cytotoxicity was noted in the absence of S9 mix at the highest concentration for TA 1535 and TA1537.

No positive mutagenic responses were observed with any of the strains used, in the presence as well as in the absence of microsomal enzymes. These results were confirmed in an independent assay.