4-(1-ethoxyvinyl)-3,3,5,5-tetramethylcyclohexanone

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-(1-ethoxyvinyl)-3,3,5,5-tetramethylcyclohexanone

EC Number:

252-961-2

EC Name:

4-(1-ethoxyvinyl)-3,3,5,5-tetramethylcyclohexanone

Cas Number:

36306-87-3

Molecular formula:

C14H24O2

IUPAC Name:

4-(1-ethoxyethenyl)-3,3,5,5-tetramethylcyclohexan-1-one

impurity 1

Reference substance name:

(3E,6E)-8-ethoxy-4,6,8-trimethylnona-3,6-dien-2-one

Molecular formula:

C14H24O2

IUPAC Name:

(3E,6E)-8-ethoxy-4,6,8-trimethylnona-3,6-dien-2-one

impurity 2

Reference substance name:

(E)-6-ethoxy-4,6,8-trimethylnona-3,7-dien-2-one

IUPAC Name:

(E)-6-ethoxy-4,6,8-trimethylnona-3,7-dien-2-one

impurity 3

Reference substance name:

(3E,6Z)-8-ethoxy-4,6,8-trimethylnona-3,6-dien-2-one

IUPAC Name:

(3E,6Z)-8-ethoxy-4,6,8-trimethylnona-3,6-dien-2-one

impurity 4

Reference substance name:

1,3-diethoxy-5,7,7-trimethylbicyclo[3.3.1]non-2-ene

IUPAC Name:

1,3-diethoxy-5,7,7-trimethylbicyclo[3.3.1]non-2-ene

impurity 5

Reference substance name:

3,5-diethoxy-1,7,7-trimethylbicyclo[3.3.1]non-2-ene

IUPAC Name:

3,5-diethoxy-1,7,7-trimethylbicyclo[3.3.1]non-2-ene

Test material form:

liquid

Specific details on test material used for the study:

Batch Number: 9000479770
Appearance: light yellow liquid
Storage: 1-10°C in the dark
Purity: 86.9%
Expiry Date: 4 Feb 2003

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Mammalian liver post--mitochondrial fraction (S-9) - prepared from male Sprague Dawley rats induced with Aroclor 1254

Test concentrations with justification for top dose:

Range finding: 1.6, 8, 40, 200, 1000 and 5000 μg/plate
Experiment 1: 1.6, 8, 40, 200, 1000 and 5000 μg/plate
Experiment 2: perfromed up to an estimate of the lower limit of toxicity 1250 μg/plate for stain TA 1537 in the absence of S-9 and strain TA102 in the absence or presence of S-9) or up to 5000 μg/plate (all other treatments). Dose ranges of 78.13 to 5000 μg/plate and 19.52 to 1250 μg/plate were employed)

Controls

Untreated negative controls:

yes

Remarks:

solvent

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: Glutaraldehyde (for TA102 without S-9), 2-aminoanthracene (for TA100, TA1535, TA1537, TA102 with S-9)

Evaluation criteria:

The test article was considered to be mutagenic if:
1. The assay was valid (see below)
2. Dunnett's test gave a significant response (p=<0.01) and the data set(s) showed a significant dose correlation
3. The positive responses described above were reproducible

Acceptance criteria
The assay was considered valid if the following criteria were met:
1. the mean negative control counts fell within the normal ranges as deinfined in Appendix 3
2. the positive control chemicals induced clear increases in revertant numbers confirming discrimination between different strains, and an active S-9 preparation
3. no more than 5% of the plates ere lost through contamination or some other unforeseen event

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

It was concluded that Kephalis did not induce mutation in five histidine-requiring strains of Salmonella typhimurium (TA98, TA100, TA1537 and TA102) when tested under the conditions of this study. The conditions included treatments at concentrations up to either 5000 μg/plate, or the lower limit of toxicity, in the absence and in the presence of a rat liver metabolic activation system (S-9).