(trans(trans))-4'-but-3-enyl-4-(4-methylphenyl)-bicyclohexyl

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 02 -April 04, 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(trans(trans))-4'-but-3-enyl-4-(4-methylphenyl)-bicyclohexyl

EC Number:

429-580-7

EC Name:

(trans(trans))-4'-but-3-enyl-4-(4-methylphenyl)-bicyclohexyl

Cas Number:

129738-42-7

Molecular formula:

Hill formula: C23H34 CAS formula: C23H34

IUPAC Name:

(1s,4r)-4-(but-3-en-1-yl)-4'-(4-methylphenyl)-1,1'-bi(cyclohexane)

Method

Target gene:

histidine for S. thyphimurium
tryptophan for E. coli

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:

Type and composition of metabolic activation system:
- source of S9 : Bioscience Research Laboratory, Kikkoman Corporation (S9; produced on Jan. 11, 1991,Lot No:RAA-248)
- method of preparation of S9 mix: A 9,000 g supernatant (S9), prepared from liver tissue of male Sprague-Dawley rats (7 weeks old) induced with phenobarbital and 5,6-benzoflavone was used. For 1 mL: 0.1 mL S9 + 8 µmol MgCl2 + 33 µmol KCl + 5 µmol G-6-P + 4 µmol NADH + 4 µmol NADPH + 0.5 mL 0.2 M Sodium phosphate buffer (pH 7.4)
- concentration of S9 mix: 0.1 mL S9 in 1 mL S9 mix
- volume of S9 mix added: 0.5 mL per plate with metabolic activation

Test concentrations with justification for top dose:

0, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

The test substance was dissolved up to 50 mg/mL in acetone, and was used for the assay immediately after diluting with the same solvent.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

furylfuramide

other: 2-aminoanthracene (2-AA)

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: duplicate
- Number of independent experiments : 1

METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar (plate incorporation)

FOR GENE MUTATION:
- Plates were incubated for 48 h at 37 °C.

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- not examined

Evaluation criteria:

The results were judged to be positive when the revertants were twice or more that of the negative control, and when dose dependency or reproducibility were observed in the increase of revertants at least in one strain.

Statistics:

The mean of two plates counted was calculated.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Remarks:

but tested up to the recommended limit concentration

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Remarks:

but tested up to the recommended limit concentration

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Remarks:

but tested up to the recommended limit concentration

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Remarks:

but tested up to the recommended limit concentration

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

Ames test:
Please refer to "Attached background material" for:
- Individual plate counts
- Mean number of revertant colonies per plate

Applicant's summary and conclusion

Conclusions:

The test item was not mutagenic in the bacterial reverse mutation study with and without metabolic activation.

Executive summary:

An in vitro bacterial reverse mutation study was conducted similar to OECD 471. In one plate incorporation test the test item was applied at concentrations of 0, 150, 500, 1500 and 5000 µg/plate to Salmonella typhimurium (TA100, TA1535, TA98, TA1537) and Escherichia coli (WP2 uvrA) strains with and without metabolic activation using S9 mix. There were no increasing number of revertants observed for any strains and any concentration of the test item. The positive control substances increased the number of mutants twice or more the negative control in each strains. Under the test conditions the test item did not show mutagenic potential in the presence and absence of metabolic activation.