reaction mass of isomers of: C7-9-alkyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP and Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: USA, EPA (TSCA) OPPTS harmonised guidelines.

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): OS 144267
- Molecular formula (if other than submission substance): C21H34O3
- Molecular weight (if other than submission substance): 334.491
- Smiles notation (if other than submission substance): Oc1c(C(C)(C)C)cc(cc1C(C)(C)C)CCC(=O)OCCCC
- InChl (if other than submission substance): 1S/C21H34O3/c1-8-9-12-24-18(22)11-10-15-13-16(20(2,3)4)19(23)17(14-15)21(5,6)7/h13-14,23H,8-12H2,1-7H3

- Structural formula attached as image file (if other than submission substance): see Fig.1
- Substance type: organic
- Physical state: yellow crystalline solid block
- Analytical purity: responsibility of the sponsor
- Lot/batch No.: no data
- Expiration date of the lot/batch:
- Stability under test conditions: responsibility of the sponsor
- Storage condition of test material: room temperature in the dark

Method

Target gene:

his- (S.thyphimurium)
trp- (E.coli)

Species / strain

Species / strain / cell type:

bacteria, other: Salmonella typhimurium TA1535, TA1537, TA98, TA100 and Escherichia coli WP2uvrA-

Additional strain / cell type characteristics:

other: S.thyphimurium: uvrB- , rfa; E.coli: uvrA-

Metabolic activation:

with and without

Metabolic activation system:

Pretreated rat liver S9

Test concentrations with justification for top dose:

Concentration range in the preliminary toxicity study (with metabolic activation): 0-5000 µg/plate
Concentration range in the range finding study and in the main test (with and without metabolic activation): 50-5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: test material is well soluble in DMSO

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

without S9-mix

Migrated to IUCLID6: 2 µg/plate for WP2uvrA-, 3 µg/plate for TA100 and 5 µg/plate for TA1535

Controls 2

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without S9-mix

Migrated to IUCLID6: 80 µg/plate for TA1537

Controls 3

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

without S9-mix

Migrated to IUCLID6: 0.2 µg/plate for TA98

Controls 4

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene (2AA): at 1 µg/plate for TA100, at 2 µg/plate for TA1535 and TA1537 and at 10 µg/plate for WP2uvrA-

Remarks:

with S9-mix

Controls 5

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

with S9-mix

Migrated to IUCLID6: at 5 µg/plate for TA98

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation).

DURATION
- Preincubation period: no
- Exposure duration: 48 hours

DETERMINATION OF CYTOTOXICITY
- Method: other: number of revertant colonies was counted and effects of the test material on the growth of bacterial background lawn was examined

OTHER: concentration of the test substance resulting in precipitation: 5000 µg/plate

Evaluation criteria:

The test material may be considered positive in this test system if the following
criteria are met:
The test material should have induced a reproducible, dose-related and
statistically (Dunnett’s method of linear regression(5)) significant increase in the
revertant count in at least one strain of bacteria.

Results and discussion

Test results

Test results 1

Species / strain:

other: TA 100 and WP2uvrA-

Metabolic activation:

with and without

Genotoxicity:

other:

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

not examined

Test results 2

Species / strain:

other: Salmonella typhimurium TA1535, TA1537, TA98, TA100 and Escherichia coli WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Observations: No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains at any dose level either with or without metabolic activation. These finding were confirmed in a total of two independent experiments.

Remarks on result:

other: other: preliminary toxicity study

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

The study was performed according to the OECD Guideline 471 without deviations and considered to be of the highest quality (reliability Klimisch 1).The vehicle and the positive control substances fulfilled validity criteria of the test system. The test material did not induced significant increases in the frequency of revertant colonies in any of the bacterial strains and therefore the test material was considered to be non-mutagenic under the conditions of the test.

Executive summary:

Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2uvrA- were treated with the test material using the Ames plate incorporation method at five dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolising system (10% liver S9 in standard cofactors).

The dose range was determined in a preliminary toxicity assay and was 50 to 5000 µg/plate in the range-finding study. The experiment was repeated on a separate day using the same dose range as the range-finding study, fresh cultures of the bacterial strains and fresh test material formulations.

The vehicle (dimethyl sulphoxide) control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with and without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9-mix were validated.

The test material caused no visible reduction in the growth of the bacterial background lawn at any dose level. The test material was, therefore, tested up to the maximum recommended dose level of 5000 µg/plate. An oily precipitate and opaque film was observed at 5000 µg/plate, this did not prevent the scoring of revertant colonies.

No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation. The test material was considered to be non-mutagenic under the conditions of this test.