(1S,4R)-1-methyl-4-(prop-1-en-2-yl)cyclohex-2-en-1-ol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019-03-14 to 2019-03-22

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(1S,4R)-1-methyl-4-(prop-1-en-2-yl)cyclohex-2-en-1-ol

Cas Number:

22972-51-6

Molecular formula:

C10H16O

IUPAC Name:

(1S,4R)-1-methyl-4-(prop-1-en-2-yl)cyclohex-2-en-1-ol

Test material form:

liquid

Details on test material:

Test item: (1S,4R)-1-Methyl-4-(prop-1-en-2-yl)cyclohex-2-enol
CAS number: 22972-51-6
Physical state: Liquid
Storage: Room temperature

Specific details on test material used for the study:

Test item: (1S,4R)-1-methyl-4-(prop-1-en-2-yl)cyclohex-2-en-1-ol (PMDOL)
CAS number: 22972-51-6
Physical state: Liquid
Storage: room temperature

Method

Target gene:

Histidine locus (histidine-dependent S. typhimurium strains)

Species / strain

Species / strain / cell type:

other: S. typhimurium: TA97a, TA98, TA100, TA102, TA1535

Details on mammalian cell type (if applicable):

not applicable

Cytokinesis block (if used):

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Rat liver enzyme solution (S9 mix)

Test concentrations with justification for top dose:

0.625 µl/plate caused no significant toxicity and was highest dose tested in the study
Dose-rangeused in study: 0.625, 0.3125, 0.15625, 0.078125 and 0.0390625 μL/plate in with and without 5% S9-mix

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine (NPD)

Remarks:

Without S9-mix; 10 μg/plate (TA97a and TA98)

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

Without S9-mix; 0.5 μg/plate (TA102)

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Without S9-mix; 0.4 μg/plate (TA100 and TA1535)

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Remarks:

Ethanol

Positive controls:

yes

Positive control substance:

other: 2-aminofluorene

Remarks:

4.0 μg/plate (TA97a with S9-mix)

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

4.0 μg/plate (TA98 with S9-mix)

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

4.0 μg/plate (TA100, TA102 and TA1535 with S9-mix)

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration (single, duplicate, triplicate): triplicate

METHOD: Plate incorporation method
The following solutions were added to 2 ml soft agar:
- 0.1 ml of a fresh bacterial culture (10^9 cells/ml) of one of the tester strains
- 0.1ml of the test article solution or control solution and
- either 0.5 ml S9-mix (in case of activation assays) or 0.5 ml 0.2 M sodium phosphate buffer (in case of nonactivation assays).
The mixtures was mixed thoroughly on a Vortex and was poured onto a minimal glucose agar plate and incubated at 37.0 °C for 48h. After this period revertant colonies were counted.

Evaluation criteria:

A test article is considered negative (not mutagenic) in the test if:
The total number of revertants in the tester strain TA 97a, TA98, TA100 and TA102 was less than twice or thrice (TA1535) the concurrent negative control (Ethanol).

Results and discussion

Test results

Key result

Species / strain:

other: TA97a, TA98, TA100, TA102, and TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

not applicable

True negative controls validity:

not applicable

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Based on the study results, no classification is proposed regarding genetic toxicity according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP).