1,4:5,8-Dimethanonaphthalene, 2-ethylidene-1,2,3,4,4a,5,8,8a-octahydro-

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:

20 February 1989 - 26 May 1989

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to official Japanese test guidelines, and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Japanese Guidelines on Industrial Chemicals (Kanpogyo No. 700, 61 Kikyoku No.1014, 1986)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Guideline under the Japanese Industrial Safety and Health Law (Notification No.77, Ministry of Labor, 1988)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(3R,6S,9E)-9-ethylidenetetracyclo[6.2.1.1³,⁶.0²,⁷]dodec-4-ene; (3S,6R,9E)-9-ethylidenetetracyclo[6.2.1.1³,⁶.0²,⁷]dodec-4-ene

EC Number:

700-518-3

Cas Number:

38233-76-0

Molecular formula:

C14H18

IUPAC Name:

(3R,6S,9E)-9-ethylidenetetracyclo[6.2.1.1³,⁶.0²,⁷]dodec-4-ene; (3S,6R,9E)-9-ethylidenetetracyclo[6.2.1.1³,⁶.0²,⁷]dodec-4-ene

Details on test material:

- Name of test material (as cited in study report): ED-TCD; also: 2-ethylidene-1,2,3,4,4a,8,8a-octahydro-1,4:5,8-dimethanonaphthalene
- Physical state: Colourless clear liquid
- Analytical purity: 99.3%
- Expiration date of the lot/batch: Stable for one year under the conditions described below
- Storage condition of test material: In a dark place at room temperature.

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 Mix

Test concentrations with justification for top dose:

Range-finding study: 0 (Solvent control), 50, 100, 500, 1000, 2000, and 5000 µg/plate.
Main test without metabolic activation, Salmonella strains: 0, 3.13, 6.25, 12.5, 25, 50, 100, 200, 400, 800 µg/plate.
Main test with metabolic activation, Salmonella strains: 0, 31.3, 62.5, 125, 250, 500, 1000 µg/plate.
Main test with and without metabolic activation, E. Coli strain: 313, 625, 1250, 2500, 5000 µg/plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulphoxide

Positive controls:

yes

Positive control substance:

other: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2)

Controls 2

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulphoxide

Positive controls:

yes

Positive control substance:

sodium azide

Controls 3

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulphoxide

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Controls 4

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulphoxide

Positive controls:

yes

Positive control substance:

9-aminoacridine

Controls 5

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulphoxide

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Controls 6

Negative solvent / vehicle controls:

yes

Remarks:

Dimethylsulphoxide

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period:20 minutes at 37°C
- Exposure duration: 48 hours at 37°C

NUMBER OF REPLICATIONS: Duplicate plates run for each


DETERMINATION OF CYTOTOXICITY
- Method: other: number of revertant colonies by comparison to the control

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: Microbial toxicity seen in TA100 and 1535 from 100 µg/plate and above without S9 mix, and from 1000 µg/plate with S9 mix. Microbial toxicity seen in TA98 and TA1537 from 500 µg/plate without S9 mix and from 1000 µg/plate with S9 mix.
No microbial toxicity seen for E Coli WP2 uvrA-, but precipitation seen from 2000 µg/plate and above.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The number of revertant colonies induced by the test substance were within the normal ranges also seen in the negative control; this suggests that the test substance is not mutagenic. Microbial toxicity (cytotoxicity) was observed in the Salmonella Typhimuium strains of bacteria tested.