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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:
1989-07-10 to 1989-09-05
Reliability:
2 (reliable with restrictions)
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:
1989
Report Date:
1989

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:
1,5-cyclooctadiene of Hüls AG. Sample ID 966/890620, purity 99.5 %

Method

Target gene:
mutated gene loci responsible for histidine auxotrophy
Species / strain
Species / strain:
other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
Additional strain characteristics:
other: histidine auxotroph
Metabolic activation:
with and without
Metabolic activation system:
Aroclor induced rat liver S9 mix, enzyme  activity tested with aminoanthracene in TA 100
Test concentrations with justification for top dose:
8; 40; 200; 1000; 5000 µg/plate
Vehicle:
Dimethylsulfoxide (DMSO)
Controls
Negative controls:
yes
Remarks:
untreated
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Remarks:
for details see below
Positive control substance:
other: without metabolic activation: Nitrofluorene (TA 98; TA 1538), Sodium azide (TA 100; TA 1535), Aminoacridine (TA 1537); with metabolic activation: Aminoanthracene (TA 100)
Details on test system and conditions:
SYSTEM OF TESTING
- Metabolic activation system:   Aroclor induced rat liver S9 mix, male Bor: WISW (SPF/Cpb) rats, enzyme  activity tested with aminoanthracene in 
TA 100
ADMINISTRATION: 
- Dosing:    
main test: 8/40/200/1000/5000 µg/plate (+/- metabolic activation)   
repeated with 157/313/625/1250/2500 µg/plate (+/- metabolic  activation), for reasons see Results, "OTHER OBSERVATIONS"   
pre-incubation test: 157/313/625/1250/2500 µg/plate (+/- metabolic  activation)
- Number of replicates: 3
- Application: solvent dimethyl sulfoxide (CAS No. 67-68-5)   
main test stock solution 50 g/l; repeat test 25 g/l   
pre-incubation test stock solution 50 g/l
- Positive and negative control groups and treatment:    
2.5 µg nitrofluorene/plate: TA 98, TA 1538 positive   
2.5 µg sodium azide/plate: TA 100, TA 1535 positive   
50 µg aminoacridine/plate: TA 1537 positive   
untreated: negative   
solvent: negative
- Pre-incubation: 30 minutes at 30 °C
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:
The stated means, standard deviations and factors were calculated through a computer program written by Messrs BIOSYS. The usual statistical
methods were used for the calculations.

Results and discussion

Test results
Species / strain:
other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
other: >= 625 µg/plate
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Remarks on result:
other:
Remarks:
Migrated from field 'Test system'.
Additional information on results:
GENOTOXIC EFFECTS: 
- With metabolic activation: none
- Without metabolic activation: none
PRECIPITATION CONCENTRATION:    
>= 1250 µg/plate (main test)   
>= 2500 µg/plate (pre-incubation test)
CYTOTOXIC CONCENTRATION (main tests):    
TA 98: none (+ S9), 1250 µg/plate (- S9)   
TA 100: none (+/- S9)   
TA 1535: 625 µg/plate (+/- S9)   
TA 1537: 1250 µg/plate (+ S9), 1000 µg/plate (- S9)   
TA 1538: 2500 µg/plate (+/- S9)
CYTOTOXIC CONCENTRATION (pre-incubation tests):    
TA 98: 1250 µg/plate (+ S9), 625 µg/plate (- S9)   
TA 100: 313 µg/plate (+/- S9)   
TA 1535: 625 µg/plate (+ S9), 313 µg/plate (- S9)   
TA 1537: 313 µg/plate (+/- S9)   
TA 1538: 1250 µg/plate (+/- S9)
OTHER OBSERVATIONS:   In the pre-incubation test, high spontaneous reverse mutation rates  were observed with and without metabolic activation 
in TA 1537 and TA  100. All other positive controls were functional. Increased toxicity  towards all strains was also observed in the 
pre-incubation test. Both  observations are tentatively assigned to the formation of peroxides  (presence of oxygen increased by shaking, 
longer residence time). Since  the pre-incubation assay could not be evaluated, a second main test was  performed.

Any other information on results incl. tables

no other remarks

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
other: negative both in presence and in absence of metabolic activation

Under the conditions of the study the test item 1,5Ccyclooctadiene proved to be non-mutagenic, both in the presence and in the absence of
Arochlor-induced liver microsomes, for all test strains used in this study.
Executive summary:

The substance 1,5-cyclooctadiene was tested in the Ames Salmonella mutagenicity test for any mutagenic activity. The test organisms were five histidine-auxotrophic Salmonella typhimurium strains (TA 1535; TA 1537; TA 1538; TA 98 and TA 100). Under the conditions of the study the test item 1,5-cyclooctadiene proved to be non-mutagenic, both in the presence and in the absence of Arochlor-induced liver microsomes, for all test strains used in this study.