(S)-2-methyl-5-(1-methylvinyl)cyclohex-2-en-1-one

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Salmonella Mutagenicity Tests: II. Results From The Testing Of 270 Chemicals

Author:

Mortelmans,K, Haworth,S, Lawlor,T, Speck,W, Tainer,B And Zeiger,E

Year:

1986

Bibliographic source:

Environ. Mutagen. 8(Suppl. 7):1 -119, 1986

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:

Gene mutation toxicity study was performed for D-Carvone to evaluate its mutagenic nature

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one

Cas Number:

2244-16-8

Molecular formula:

C10H14O

IUPAC Name:

(5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one

Details on test material:

- Name of test material: D-Carvone- IUPAC name: (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one- Molecular formula: C10H14O- Molecular weight: 150.22 g/mol- Substance type: Organic

Specific details on test material used for the study:

- Name of test material: D-Carvone- IUPAC name: (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one- Molecular formula: C10H14O- Molecular weight: 150.22 g/mol- Substance type: Organic- Physical state: No data- Purity: 95.4%- Impurities (identity and concentrations): 4.6%

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

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

Remarks:

Lab 1 and Lab 2

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

Male Sprague-Dawley rats and male Syrian hamsters were routinely used for the S9 preparation of the liver fractions

Test concentrations with justification for top dose:

0, 3.3, 10.0, 33.0, 100.0, 333.0 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO- Justification for choice of solvent/vehicle: The test chemical was 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:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine (TA98; -S9), 2-aminoanthracene (all strains, +S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubationDURATION- Preincubation period: 20 mins- Exposure duration: 48 hr- Expression time (cells in growth medium): 48 hr- Selection time (if incubation with a selection agent): No data- Fixation time (start of exposure up to fixation or harvest of cells): No dataSELECTION AGENT (mutation assays): No dataSPINDLE INHIBITOR (cytogenetic assays): No dataSTAIN (for cytogenetic assays): No dataNUMBER OF REPLICATIONS: At least five dose levels of the chemicals were tested, with three plates per dose level.NUMBER OF CELLS EVALUATED: No dataDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No dataOTHER EXAMINATIONS:- Determination of polyploidy: No data- Determination of endoreplication: No data- Other: No dataOTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

1) mutagenic response: a dose-related, reproducible increase in the number of revertants over background, even if the increase was less than twofold; 2) nomutagenic response: when no increase in the number of revertants was elicited by the chemical; 3) questionable response: when there was an absence of a clear-cut dose-related increase in revertants; when the dose-related increases in the number of revertants were not reproducible;or when the response was of insufficient magnitude to support a determination of mutagenicity

Statistics:

Mean and Standard error of mean

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA100, TA1535, TA1537, TA98

Remarks:

Lab 1 and Lab 2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS- Effects of pH: No data - Effects of osmolality: No data- Evaporation from medium: No data- Water solubility: No data- Precipitation: No data- Other confounding effects: No dataRANGE-FINDING/SCREENING STUDIES: The chemical was initially tested with strain TA100 in the presence and the absence of the metabolic activation systems, over a wide dose range with an upper limit of 10 mg/plate, or less when solubility problems were encountered. Toxicity was evidenced by one or more of the following phenomena: appearance of his+ pinpoint colonies, reduced numbers of revertant colonies per plate, or thinning or absence of the bacterial lawn. Nontoxic chemicals were tested in the initial experiment up to the 10 mg/plate dose level, or to a level determined by their solubility. Toxic chemicals were tested up to a high dose which exhibited some degree of toxicity.COMPARISON WITH HISTORICAL CONTROL DATA: No data

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Mutation data for the test chemical D- carvone

Dose (µg/plate)	TA100
	NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM
0	89	4.6	121	12.1	133	11.7
3.3	84	7.6	119	9.9	94	14.1
10.0	75	4.0	113	9.6	105	6.7
33.0	89	4.1	107	6.7	108	2.2
100.0	73	7.4	104	6.7	108	2.2
333.0	73	7.4	104	3.0	100	5.0
Positive control	48	9.0	78	8.4	65	11.3

 

Dose (µg/plate)	TA1535
	NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM
0	4	1.5	6	1.5	8	3.0
3.3	3	0.9	5	1.3	5	0.3
10.0	3	0.7	4	0.9	5	1.2
33.0	3	0.	3	0.7	4	1.5
100.0	2	1.0	5	0.3	3	0.6
333.0	1	0.6	4	0.6	3	2.2
Positive control	122	11.1	41	5.8	76	13.4

 

Dose (µg/plate)	TA1537
	NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM
0	3	0.7	6	0.6	6	1.3
3.3	3	0.6	5	0.7	5	1.2
10.0	2	0.0	5	1.2	8
33.0	3	1.2	2	0.7	7	1.7
100.0	3	1.0	4	0.7	8	2.7
333.0	3	0.6	6	1.2	6	0.9
Positive control	1041	154.2	261	12.0	211	24.1

 

Dose (µg/plate)	TA98
	NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM
0	14	1.9	25	1.2	21	3.8
3.3	16	2.8	23	5.0	20	3.2
10.0	15	0.3	23	1.2	15	3.2
33.0	11	1.2	25	2.5	22	0.9
100.0	13	0.9	23	2.0	24	1.5
333.0	10	2.1	17	4.0	22	2.0
Positive control	359	21.4	1969	56.9	973	201.3

Applicant's summary and conclusion

Conclusions:

D- carvone did not induce gene mutation in the Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

Executive summary:

Gene mutation toxicity study was performed for D- carvone to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of0, 3.3, 10.0, 33.0, 100.0, 333.0 µg/plate. DMSO was used at the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. D- carvone did notinduce gene mutation in theSalmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.