3,5,5-trimethylcyclohex-2-enone

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:

guideline study

Remarks:

Restriction: only 4 Salmonella strains tested

Data source

Reference

Reference 1

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, Zeiger E

Year:

1986

Bibliographic source:

Environ Mutagen 8 Suppl. 7: 1-119

Reference 2

Reference Type:

review article or handbook

Title:

Toxicology and Carcinogenesis Studies of Isophorone in F344/N Rats and B6C3F1 Mice.

Author:

NTP (U.S. National Toxicology Program)

Year:

1986

Bibliographic source:

Techn Rep Ser No. 291, U.S. Department of Health and Human Services

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

4 strains tested instead of 5

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3,5,5-trimethylcyclohex-2-enone

EC Number:

201-126-0

EC Name:

3,5,5-trimethylcyclohex-2-enone

Cas Number:

78-59-1

Molecular formula:

C9H14O

IUPAC Name:

3,5,5-trimethylcyclohex-2-enone

Details on test material:

Purity ≥ 97%

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 rat or hamster liver S9

Test concentrations with justification for top dose:

TA 100 and TA 1535 : 0 - 10000 μg/plate
TA 98 and TA 1537: 0 - 3333 μg/plate

Vehicle / solvent:

water

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Remarks:

potassium chloride

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Strains TA 1535 and TA 100, without metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Remarks:

potassium chloride

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine

Remarks:

Strain TA 98, without metabolic activation

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Remarks:

potassium chloride

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Strain TA 1537, without metabolic activation

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Remarks:

potassium chloride

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

All strains, with metabolic activation

Details on test system and experimental conditions:

- Number of replicates: 3 per dose level, repeated
- Solvent: H2O
- 3 investigations were performed:
1. without metabolic activation
2. with metabolic activation (Arochlor-1254 liver rats)
3. with metabolic activation (Arochlor-1254 liver hamster)
- Cells and test compound or solvent (water) were incubated for 20 minutes at 37 ºC in the presence of either S9 or buffer. After the addition of soft agar, the contents of each tube were poured onto minimal medium, and the plates were incubated at 37 °C for 48 hours.
- The analysis was performed twice, each in triplicates.

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:

other: -S9: TA100, 1535, 98 at concentrations >=1000 µg/plate; +S9 (rat and hamster): only TA1535 at 10000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The test substance was not mutagenic in strains TA100, TA1535, TA1537, or TA98 of Salmonella Typhimurium in the presence or absence of Aroclor 1254-induced rat (r.) or hamster (h.) liver S9.
Differences were not significant.

Maximum number of revertants in the standard plate test:

TA100:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
-----------------------------------------------------------
82±4.7 92±7.5 79±2.2 88±7.4(333) 90±2.2(100) 117±2.9(333)

TA1535:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
---------------------------------------------------------
6±2.5 7±0.9 7±1.2 4±0.9(100) 9±2.8(333) 6±1.0(100)

TA1537:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
----------------------------------------------------------
2±0.3 3±1.5 4±0.7 4±0.7(1000) 5±1.0(100) 6±2.0(1000)

TA98:
Vehicle Test substance (µg/plate)
-S9 +S9(r.) +S9(h.) -S9 +S9(r.) +S9(h.)
----------------------------------------------------------
10±2.0 11±2.9 17±1.0 12±2.3(100) 17±1.3(100) 15±0.9(333)

The analysis was performed twice, each in triplicate; because the results were similar, data from only one experiment are shown.