N-ethyl-2-(isopropyl)-5-methylcyclohexanecarboxamide

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:

secondary literature

Justification for type of information:

Data from peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Mutagenic investigation of test chemical in the Salmonella/mammalian microsome test

Author:

Pia Haubro Andersen and et. al.

Year:

1984

Bibliographic source:

Mutation Research (1984)

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Salmonella Mutagenicity Tests of test chemical was performed by Bacterial gene mutation assay

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

trans-menthone

EC Number:

201-941-1

EC Name:

trans-menthone

Cas Number:

89-80-5

Molecular formula:

C10H18O

IUPAC Name:

(2S,5R)-2-Isopropyl-5-methylcyclohexanone

Test material form:

solid

Details on test material:

IUPAC name:2-Isopropyl-5-methylcyclohexanone
Mol. formula: C10H18O
Molecular Weight: 154.2512
Smiles: C1([C@@H](CC[C@@H](C1)C)C(C)C)=O
InChI: 1S/C10H18O/c1-7(2)9-5-4-8(3)6-10(9)11/h7-9H,4-6H2,1-3H3

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

No data available

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data available

Metabolic activation:

with and without

Metabolic activation system:

(The S-9 fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared)

Test concentrations with justification for top dose:

0.00, 6.4, 32, 160, 800 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Test material soluble in DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-nitrofluorene

sodium azide

other: 2-anthramine for all strains with S9 metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION:
Standard plate
DURATION
- Preincubation period: No data available

- Exposure duration: No data available
- Expression time (cells in growth medium): No data available
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: 5 plate

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available
OTHER EXAMINATIONS:
- Determination of polyploidy: No data available-
Determination of endoreplication: No data available-
Other: No data available

OTHER: No data available

Rationale for test conditions:

No data available

Evaluation criteria:

The histidine-revertant (his') colonies arising on the plates were counted

Statistics:

Mean number of revertants for n mumber of plates for each dose level was calculated to be the square value of mean(y) of squared roots. Standard error of mean was calculated

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA98, TA100, TA1535 ,TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Remarks:

DMSO

Untreated negative controls validity:

valid

Remarks:

DMSO

Positive controls validity:

valid

Additional information on results:

No data available

Any other information on results incl. tables

Dose µg/plate	TA1535			TA100		TA1538		TA98
	Without S9		With S9	Without S9	With S9	Without S9	With S9	Without S9	With S9
0.00	7±1	8±1		85±4	114±8	7±1	11±1	32±3	33±2
6.4	7±1	10±1		96±4	114±8	13±1	11±1	25±2	28±2
32	7±1	9±1		92±4	121±8	12±2	12±1	31±3	27±2
160	5±1	8±1		97±4	123±8	8±2	11±1	28±2	26±2
800	7±1	9±1		69±6	104±7	4±1	11±1	13±3	22±2
Sodium Azide	499±20			361± 20
2-Anthramine		169± 7			278 ±12		119± 6		233± 13
2-Nitrofluorene						38± 3		186± 13

Applicant's summary and conclusion

Conclusions:

Salmonella Mutagenicity Tests of test chemical was performed in Salmonella strain TA98, TA100, TA1535, and TA 1537 did not produce mutation, therefore it is considered to be negative for gene mutation in vitro.

Executive summary:

SalmonellaMutagenicity Tests of test chemical was performed in Salmonella strainTA98, TA100, TA1535, and TA 1537. Both in the presence and absence of S9 metabolic activation system.

 

 The test chemical was dissolved in DMSO to made dose concentration of 0.00, 6.4, 32, 160, 800 µg/plate in 5 parallel plates.The S-9 fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared.However 2-anthramine served positive control with S9 fraction for all strain of Salmonella typhimurium and sodium azide (TA1535 and TA 100), 2-nitrofluorene (TA 1538 and TA98) served as positive control without S9 fraction.

 

Mutagenicity evaluated by counting histidine-revertant (his') colonies arising on these plates. As test chemical did not produce mutationin Salmonella strainTA98, TA100, TA1535, and TA 1537,Therefore itis considered to be negative for gene mutation in vitro.