Menthane, monohydroperoxy derivative

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: standard protocol, non GLP, tables available on results of the test substance

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1988

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

A test for the ability of the test substance to induce mutations in strains of Salmonella typhimurium was made. Concurrent solvent and positive controls were run with each trial. Histidine-independent (his+) colonies arising on these plates were counted following two days incubation.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

p-menthane hydroperoxide

IUPAC Name:

p-menthane hydroperoxide

Constituent 2

Reference substance name:

1-methyl-1-(4-methylcyclohexyl)ethyl hydroperoxide

EC Number:

201-281-4

EC Name:

1-methyl-1-(4-methylcyclohexyl)ethyl hydroperoxide

Cas Number:

80-47-7

IUPAC Name:

1-methyl-1-(4-methylcyclohexyl)ethyl hydroperoxide

Constituent 3

Reference substance name:

Menthane, monohydroperoxy derivative

EC Number:

247-987-6

EC Name:

Menthane, monohydroperoxy derivative

Cas Number:

26762-92-5

Molecular formula:

C10H20O2

IUPAC Name:

menthane, monohydroperoxy derivative

Details on test material:

Purity: label purity: 55%
Batch No.: not specified

Method

Target gene:

Histidine

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of culture media: Oxoid No.2 broth
- Properly maintained: yes

No additional data

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 97

Details on mammalian cell type (if applicable):

- Type and identity of culture media: Oxoid No.2 broth
- Properly maintained: yes

No additional data

Metabolic activation:

with and without

Metabolic activation system:

rat and hamster S-9 mix

Test concentrations with justification for top dose:

TA100 without S9: 0, 1, 3, 10, 33, 100 μg/plate
TA100 with S9: 0, 3, 10, 33, 100, 333 μg/plate (10% HLI); 0, 10 33, 100, 333, 666 μg/plate (10% RLI)
TA 1535 without S9: 0, 0.3, 1, 3, 10, 16 μg/plate
TA 1535 with S9: 0, 3, 10, 33, 100, 333 μg/plate (10% HLI); 0, 10, 33, 100, 333, 666 μg/plate (10% RLI)
TA 1537 without S9: 0, 1, 3, 10, 33, 66 μg/plate
TA 1537 with S9: 0, 10, 33, 100, 166, 333 μg/plate (5, 10 and 30% RLI)
TA 97 without S9: 0, 1, 3, 10, 33, 66 μg/plate
TA 97 with S9: 0, 3, 10, 33, 100, 333 μg/plate (10% HLI); 0, 10, 33, 100, 166, 333 μg/plate (5, 10 and 30% RLI)
TA 98 without S9: 0, 1, 3, 10, 33, 100 μg/plate
TA 98 with S9: 0, 3, 10 33, 100, 333 μg/plate (10% HLI); 0, 10, 33, 100, 333, 666 μg/plate (10% RLI)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

No additional data.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

In the absence of S9 mix for strains TA1535 and TA100

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

In the absence of S9 mix for strains TA97 and TA 1537

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine

Remarks:

In the absence of S9 mix for strains TA98

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

In the presence of S9 mix for all strains

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: The test substance (0.05 mL), Salmonella culture (0.10 mL) and S-9 mix or buffer (0.50 mL) were incubated at 37 ℃, without shaking, for 20 min.
- Exposure duration: 2 days

NUMBER OF REPLICATIONS: 1 without S9; 1-3 with different concentration of S9

No additional data.

Evaluation criteria:

A chemical was judged mutagenic (+) or weakly mutagenic (+W) if it produced a reproducible dose-related response over the solvent control in replicate trails. A chemical was judged questionable (?) if the result of individual trials were not reproducible, if increases in his+ revertants did not meet the criteria for a “+W” response, or if only single dose produced increases in his+ revertants in repeat trials. Chemicals were judged nonmutagenic (-) if they did not meet the criteria for a mutagenic of questionably response.

Statistics:

None stated.

Results and discussion

Test results

Test results 1

Key result

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:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

ambiguous

Remarks:

positive with metabolic activation, negative without metabolic activation

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
The toxicity assay was performed using TA100 indicating a slight reduction of bacterial background lawn at 100 and 666 μg/plate (without and with S9 resp.)

Any other information on results incl. tables

No dose dependent increase in the number of His+ revertants was observed in TA100, TA1535, TA1537 and TA98 with or without metabolic activation

No dose dependent increase in the number of His+ revertants was observed in TA97 without metabolic activation

A dose dependent increase in the number of His+ revertants was observed in TA97 with metabolic activation (rat S9 only), not reaching a two-fold increase compared to DMSO controls at the highest tested concentration

Applicant's summary and conclusion

Conclusions:

Interpretation of results
negative without metabolic activation
ambiguous with metabolic activation

The test substance did not induce mutations in Salmonella typhimurium strains without metabolic activation
With metabolic activation the results of the Salmonella typhimurium assay with test substance were ambigeous.

Executive summary:

The test substance was tested for its ability to induce mutations in strains of Salmonella typhimurium. Concentrations ranged from 0 -333 μg/plate (cytotoxicity at 666 μg/plate)

Concurrent solvent and positive controls were run with each trial. The histidine-independent (his+) colonies arising on these plates were counted following two days incubation.

The test substance did not induce mutations in Salmonella typhimurium strains without metabolic activation

With metabolic activation the results of the Salmonella typhimurium assay with test substance were ambigeous.