Registration Dossier

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
The study was conducted in 1980.
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Study is well conducted but its design is not sufficient to cover the endpoint.

Data source

Reference
Reference Type:
publication
Title:
SCREENING OF TOBACCO SMOKE CONSTITUENTS FOR MUTAGENICITY USING THE AMES' TEST.
Author:
Florin I et al.
Year:
1980
Bibliographic source:
Toxicology 18 (1980) 219 – 232.

Materials and methods

Test guideline
Qualifier:
no guideline followed
Deviations:
not applicable
Principles of method if other than guideline:
Substances were assayed for mutagenicity towards four histidine-requiring mutants of Salmonella typhimurium (TA 98, TA 100, TA1535 and TA 1537). The test was performed with and without metabolic activation using a liver fraction (S-9) from Aroclor 1254 of methylchloanthrene induced rats using a modified Ames test. Due to the large number of compounds screened during the study potential mutagens were screened using spot tests which are less sensitive than quantitative experiments.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
6-isopropyl-3-methylcyclohex-2-enone
EC Number:
201-942-7
EC Name:
6-isopropyl-3-methylcyclohex-2-enone
Cas Number:
89-81-6
Molecular formula:
C10H16O
IUPAC Name:
3-methyl-6-(propan-2-yl)cyclohex-2-en-1-one
Test material form:
not specified
Details on test material:
Test substance: Piperitone
Purity: > 97 %

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
3 µmol/plate
Vehicle / solvent:
Ethanol.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
no
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
other: 2-aminoanthracene
Details on test system and experimental conditions:
Revertants were scored on glucosen minimal salts medium supplemented with 0.05 µmol histidine and 0.05 µmol biotin. Plates used for viable counts contained 10 µmol histidine (and 0.05 µmol biotin).

The following controls were made for each experiment
The viable count was determined.
The number of spontaneous revertants was measured.
The presence of the rfa-mutation was checked by crystal violet inhibition.
The presence of the plasmid pKM 101 in strains TA98 and TA100 was checked by resistance to ampicillin.
The response of the positive controls, N-methyl-N'-nitro-N-nitrosoguanidin (not requiring metabolic activation) and 2-aminoanthracene (requiring activation) was checked.

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:
no cytotoxicity
Vehicle controls validity:
not examined
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
The test substance was not found to be mutagenic.
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):
negative

The test substance was not found to be mutagenic.
Executive summary:

The genotixic potential of the test substance was assessed using a modified Ames test in which potential mutagens were screened using spot tests. The test substance was not found to be mutagenic under the conditions of the test, both with and without metabolic activation.