4-sec-butyl-2,6-di-tert-butylphenol

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:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

not specified

GLP compliance:

not specified

Remarks:

GLP compliance yet to be confirmed.

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-sec-butyl-2,6-di-tert-butylphenol

EC Number:

241-533-0

EC Name:

4-sec-butyl-2,6-di-tert-butylphenol

Cas Number:

17540-75-9

Molecular formula:

C18H30O

IUPAC Name:

4-(butan-2-yl)-2,6-di-tert-butylphenol

Test material form:

liquid

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

Dose setting test, with and without S9-mix, (TA1535 TA1537,TA98 ,TA100 and WP2uvrA): 5, 15, 50, 150, 500, 1500*, 5000* µg/plate
Experiment 1, without S9-mix, (TA1535 TA1537,TA98 ,TA100 and WP2uvrA):156, 313, 625, 1250, 2500*, 5000* µg/plate
Experiment 1, with S9-mix, (TA1535 TA1537,TA98 ,TA100 and WP2uvrA):156, 313, 625*, 1250*, 2500*, 5000* µg/plate

*precipitation occured

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2)

Remarks:

Without S9-mix, TA100 and Wp2uvrA: 0.01 µg/plate; TA98 0.1 µg/plate

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Without S9-mix, TA1535: 0.5 µg/plate

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Without S9-mix, TA1537: 80 µg/plate

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (2-AA)

Remarks:

With S9-mix, TA100: 1 µg/plate, TA98: 0.5 µg/plate, WP2uvrA: 10 µg/plate, TA1537 and TA1535: 2 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: pre-incubation method

NUMBER OF REPLICATIONS:
- Doses of the test substance were tested in triplicate in each strain.

OTHER EXAMINATIONS:
- The presence of precipitation and cytotoxicity of the test compound on the plates was determined.

Evaluation criteria:

The experimental results were judged positive when the number of revertant colonies was twice or more that of the negative control and showed a dose-relationship.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Precipitation occured in the dose setting experiment, with and without S9-mix, at dose levels 2500 and 5000 µg/plate.
Precipitation occured in the main experiment, without S9-mix, at dose levels 2500 and 5000 µg/plate.
Precipitation occured in the main experiment, with S9-mix, at dose levels 625, 1250, 2500, 5000 µg/plate

No toxicity was observed during the experiments.

Any other information on results incl. tables

VALIDITY

The number of revertants in negative control groups were comparible to in house historical data.

The number of revertants in the positive control showed an increase more than 2 times compared to the negative control groups.

Applicant's summary and conclusion

Conclusions:

In an AMES test, 4-sec-butyl-2,6-Di-butylphenol was found to be not mutagenic with or without metabolic activation in all five tested strains.