Cycloocta-1,5-diene

Genetic toxicity: in vitro

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

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

Test material information

Constituent 1

Reference substance name:

Cycloocta-1,5-diene

EC Number:

203-907-1

EC Name:

Cycloocta-1,5-diene

Cas Number:

111-78-4

Molecular formula:

C8H12

IUPAC Name:

(1Z,5Z)-cycloocta-1,5-diene

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 / cell type:

other: TA 98, TA 100, TA 1535, TA 1537, TA 1538

Additional strain / cell type 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 / solvent:

Dimethylsulfoxide (DMSO)

Controls

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle 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 experimental 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:

S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: >= 625 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

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.

Remarks on result:

other: other: Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98, TA 100

Remarks:

Migrated from field 'Test system'.

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.