Allyl 3-cyclohexylpropionate

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Non-GLP study similar to OECD TG 471, only four relevant strains are tested

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1983

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Only four relevant strains tested

Principles of method if other than guideline:

A standard plate assay was followed according to Ames et al. (1975): Ames BN, McCann J and Yamasaki E, Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutation Research 31, 347-363, 1975

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Allyl 3-cyclohexylpropionate

EC Number:

220-292-5

EC Name:

Allyl 3-cyclohexylpropionate

Cas Number:

2705-87-5

Molecular formula:

C12H20O2

IUPAC Name:

prop-2-en-1-yl 3-cyclohexylpropanoate

Method

Target gene:

Histidine

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:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

S9-mix from Aroclor-pretreated rats

Test concentrations with justification for top dose:

5 concentrations tested per strain, no details on concentrations provided, concentration up to 3.6 mg/plate tested

Vehicle / solvent:

Dimethylsulphoxide may have been used as solvent

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: soidum azide, benzo[a]pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation), Vogel-Bonner medium was used

DURATION
- Preincubation period: if applicable, then liquid pre-incubation for 20 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 2

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

Positive result if 1) statistical significance, 2) a reproducible, dose-related and at least two-fold elevation of the spontaneous revertant frequency; agents producing reproducible, dose-related and significant (p equal to or less than 0.01) but less than two-fold elevations were classified as marginally mutagenic

Statistics:

Statistical significance in difference between test groups and negative controls was determined according to the methods of Kastenbaum and Bowman (Tables for determining the statistical significance of mutation frequencies, Mutation Research 9, 527-549, 1970)

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:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Test results 2

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Test results 3

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Test results 4

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

No further details on results reported

Any other information on results incl. tables

Number of revertants in positive controls with sodium azide: 400 to 700 in TA100, 430 to 760 in TA1535

Number of revertants in positive controls with benzo[a]pyrene: 660 to 1000 in TA98, 865 to 1210 in TA100, 235 to 350 in TA1537, 410 to 590 in TA1538

Applicant's summary and conclusion

Conclusions:

The substance was non-mutagenic in an Ames test with Salmonella typhimurium in the absence and presence of metabolic activation (S9-mix).

Executive summary:

The potential of the substance allyl 3-cyclohexylproprionate to cause reverse mutations in bacteria (Salmonella typhimurium) was studied in a non-GLP Ames test with the strains TA98, TA100, TA1535, TA1537 and TA1538. The standard plate testing procedure was followed (Ames et al. 1975) that was widely similar to OECD TG 471, with the exception that only four relevant bacterial strains were tested. Overnight bacterial cultures had cell titres of at lest 10E09 cells/mL. S9-mix was prepared from Aroclor-pretreated rats (intraperitoneal injection of 500 mg/kg bw) and adjusted to 25 mg protein/mL. An aliquot of 0.5 mL S9-mix equivalent to 50 µL S9 -mix was incorporated into the plates. Dimethylsulphoxide was used as solvent for test substances that were poorly soluble in water. Five doses of the substance were tested (up to 3.6 mg/plate) in all five tester strains with and without S9-mix. Vogel-Bonner medium was used and plates were incubated for 48 hours. Positive controls were run in parallel with the reference substances sodium azide (0.5 µg/plate tested with TA100 and TA1535) and benzo[a]pyrene (5 µg/plate tested with TA98, TA100, TA1537 and TA1538). The substance was tested at least twice. The substance allyl 3-cyclohexylproprionate was not mutagenic in any of the tested bacterial strains in the absence and presence of metabolic activation system under the conditions of the test.