Cineole

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:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment.

Data source

Reference

Reference Type:

publication

Title:

Salmonella Mutagenicity Test Results for 250 Chemicals

Author:

Haworth S et al.

Year:

1983

Bibliographic source:

Haworth S, Environmental Mutagenesis Supplement 1:3-142 (1983)

Report date:

1983

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Cineole

EC Number:

207-431-5

EC Name:

Cineole

Cas Number:

470-82-6

Molecular formula:

C10H18O

IUPAC Name:

1,3,3-trimethyl-2-oxabicyclo[2.2.2]octane

Constituent 2

Reference substance name:

Eucalyptol

IUPAC Name:

Eucalyptol

Details on test material:

- Name of test material (as cited in study report): Cineol (Eucalyptol)
- Lot/batch No.: C22435
- Other: Sourced from Pfaltz and Bauer

Method

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced liver S-9 fractions obtained from male Sprague-Dawley rats and male Syrian hamsters, injected, i.p.

Test concentrations with justification for top dose:

The maximum concentration dosed was 10mg/plate unless toxicity was observed at this dose level.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: [acetone; dimethyl sulphoxide; ethanol(95%); water (distilled)]
- Justification for choice of solvent/vehicle: The solvent of choice was distilled water, DMSO was used if the chemical was insoluble. Ethanol or acetone was used if the substance was not soluble or stable in DMSO. The final choice of solvent for this substance was not reported.

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Tested on TA1537

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Tested on TA100 and TA 1535 without S9.

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-Nitro-o-phenylenediamine (NOPD)

Remarks:

Tested on TA98 without S9

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene (2-AA)

Remarks:

Used on all strains with and without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation procedure
DURATION
- Preincubation period: 37°C for 48 hours in the open lab (without shaking)

SELECTION AGENT (mutation assays):

NUMBER OF REPLICATIONS: At least five doses of test substance, in addition to the concurrent solvent and positive controls were tested on each strain in the presence of S-9 mix or buffer. Three plates were used, and the experiment was repeated no less than 1 week after completion of the initial test.

NUMBER OF CELLS EVALUATED: Not reported

DETERMINATION OF CYTOTOXICITY
- Method: viability on complete medium (EGG) and reduced numbers of revertant colonies per plate and/or thinning or the absence of the bacterial lawn (CWR, EGG, SRI). If toxicity was not apparent in the preliminary toxicity determination, the highest dose tested was 10mG/plate; otherwise the upper limit of solubility was used. If toxicity was observed, the doses of test chemical were chosen so that the high dose exhibited some degree of toxicity.
OTHER:

Evaluation criteria:

Positive controls: The positive contol chemicals were tested concurrently with each test chemical. 2-AA was tested on all strains in the presence of rat and hamster S-9. NOPD was tested on TA98 with out S9. SA was tested on TA100 and TA1535 and 9-AAD was tested on TA1537 all without S9. The concentration for each poistive control used for each strain and activation condition was selected based on dose-response curves gebnerated at the beginning of the test. All positive controls were detected and gave reproducible results.
The negative controls were non-mutagenic in all tests.

Statistics:

The data were evaluated using analysis based on the models presented by Margolin et al [1981].

Results and discussion

Test results

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:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
other: negative with and without metabolic activation system

The test substance was tested blind following the Ames test method (reverse phase mutagenicity test) using strains TA1535, TA1537, TA98 and TA100, with and without S9. The substance did not exhibit any mutagenic effects on the strains tested with or without S9.
Concurrent positive control samples gave expected and reproducible effects with and without S9 under the conditions of the test, thereby confirming the satisfactory activity of the metabolic activation system used.
Negative(solvent) controls gave expected and reproducible responses under the conditions of the test.

Executive summary:

The test substance was tested blind following the Ames test method (reverse phase mutagenicity test) using strains TA1535, TA1537, TA98 and TA100, with and without S9. The substance did not exhibit any mutagenic effects on the strains tested with or without S9.

Concurrent positive control samples gave expected and reproducible effects with and without S9 under the conditions of the test, thereby confirming the satisfactory activity of the metabolic activation system used.

Negative(solvent) controls gave expected and reproducible responses under the conditions of the test.