Reaction mass of 7,7-dimethyl-2-methylidenebicyclo[2.2.1]heptane and (1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane and (1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane and (1S)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(Only two of the recommended strains were tested)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Pin-2(3)-ene

EC Number:

201-291-9

EC Name:

Pin-2(3)-ene

Cas Number:

80-56-8

Molecular formula:

C10H16

IUPAC Name:

2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

Method

Target gene:

Histidine-requiring gene in Salmonella typhimurium

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 100

Additional strain / cell type characteristics:

other: uvrB and rfa mutated

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 98

Additional strain / cell type characteristics:

other: uvrB and rfa mutated

Species / strain 3

Species / strain / cell type:

S. typhimurium, other: UTH 8414

Species / strain 4

Species / strain / cell type:

S. typhimurium, other: UTH 8413

Metabolic activation:

with and without

Metabolic activation system:

liver homogenate (S9) from Aroclor-induced male Sprague-Dawley rats (100 µL/plate)

Test concentrations with justification for top dose:

Five concentrations, from 10 µg/plate to 500 µg/plate.
Toxicity and/or solubility determined the upper limit of the dose tested.

Vehicle / solvent:

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

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

(With and without metabolic activation)

Controls 2

Positive controls:

yes

Positive control substance:

sodium azide

Controls 3

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene

Controls 4

Positive controls:

yes

Positive control substance:

other: Cisplatin

Details on test system and experimental conditions:

DURATION
- Exposure duration: The plates were incubated at 37 ºC for 48 h, at which time the number of colonies per plate was determined.

SELECTION AGENT (mutation assays): the lack of amino-acid in the medium. Only the mutants can grow due to their capability to synthesize the essential amino acid.

NUMBER OF REPLICATIONS: 2

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

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:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

other: UTH 8414

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

other: UTH 8413

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Alpha pinene was not mutagenic in all strains tested with and without metabolic activation.

Applicant's summary and conclusion

Conclusions:

Alpha pinene was not mutagenic in all strains tested with and without metabolic activation.

Executive summary:

Alpha pinene was tested for mutagenecity (doses: 10 -500µg/plate). The mutagenicity assay employed was that described by Maron and Ames using Salmonella typhimurium strains TAl00 and TA98, which are DNA-repair deficient, and two additional strains, UTH 8414 and UTH 8413, developed by Matney, which have full DNA repair capacity. The mutagenicity assays were carried out both with and without metabolic activation (S9). Alpha pinene was not mutagenic in all strains tested with and without metabolic activation.