Methyl 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:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: non-guideline study, non-GLP study, no E.Coli WP2 or TA 102 was tested, with QAU statement, comparable to guideline, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no E.coli WP2 or Ta 102 strains were tested, no details on cytotoxicity, no historical data, overall limited details

GLP compliance:

no

Remarks:

QAU statement included

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate

EC Number:

228-985-4

EC Name:

Methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate

Cas Number:

6386-38-5

Molecular formula:

C18H28O3

IUPAC Name:

methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propanoate

Details on test material:

- Analytical purity: > 99%

Method

Target gene:

his

Species / strain

Species / strain / cell type:

other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced S9 fraction of rat liver

Test concentrations with justification for top dose:

20, 80, 320, 1280 and 5120 µg/0.1 ml in two independent experiments.

Vehicle / solvent:

- Vehicle used: acetone

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: see "details on test system"

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
The experiment was performed twice to confirm the results.

DURATION
- Exposure duration: 48h at 37°C

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- growth of background lawn

POSITIVE CONTROLS
- Without S9-mix
TA 98: daunorubicin-HCl, 5 and 10 µg/0.1 ml phosphate buffer;
TA 100: 4-nitroquinoline-N-oxide, 0.125 and 0.25 µg/0.1 ml phosphate buffer
TA 1535: N-methyl-N'-nitro-N-nitrosoguanidine, 3 and 5 µg/0.1 ml phosphate buffer
TA 1537: 9(5)aminoacridine hydrochloride monohydrate, 50 and 100 µg/0.1 ml DMSO
TA 1538: 2-nitrofluorene, 5 and 10 µg/0.1 ml DMSO.
- With S9-mix
TA 1535: cyclophosphamide, 250 µg/0.1 ml phosphate buffer.

Evaluation criteria:

The test substance is generally considered to be nonmutagenic if the colony count in relation to the negative control is not doubled at any concentration.

Statistics:

When the colonies had been counted, the arithmetic mean was calculated.

Results and discussion

Test results

Species / strain:

other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

- Precipitation: At the concentration of 5120 ug/0.1 ml the substance precipitated in soft agar.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

NUMBER OF BACK-MUTANT COLONIES PER PLATE (ARITHMETIC MEAN)

EXPERIMENT 1

	TA 98		TA 100		TA 1535		TA 1537		TA 1538
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
Dose/0.1 ml
Solvent Control	19	38	185	173	10	15	5	14	13	26
20 µg	21	45	178	157	12	12	6	13	8	23
80 µg	18	40	188	171	15	14	9	13	9	22
320 µg	21	42	156	175	12	12	9	13	9	26
1280 µg	20	34	192	167	8	15	5	15	7	24
5120 µg	22	38	160	151	13	9	4	16	11	15
Positive Controls*
Solvent Control	26	-	179	-	8	11	7	-	14	-
Concentration A	534	-	657	-	395	455	103	-	731	-
Concentration B	880	-	1063	-	1394	-	724	-	968	-

EXPERIMENT 2

	TA 98		TA 100		TA 1535		TA 1537		TA 1538
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
Dose/0.1 ml
Control	23	42	155	158	13	11	6	9	12	28
20 µg	18	41	158	138	10	12	9	12	14	21
80 µg	22	41	135	124	9	11	5	12	11	24
320 µg	26	46	150	148	11	12	7	13	10	18
1280 µg	22	37	147	143	10	14	6	15	12	23
5120 µg	17	29	128	123	11	10	6	11	10	24
Positive Controls*
Control	15	-	128	-	12	10	9	-	13	-
Concentration A	377	-	627	-	112	525	45	-	888	-
Concentration B	605	-	989	-	772	-	219	-	1662	-

* Details on positive controls including concentrations see "Details on test system and conditions"

Applicant's summary and conclusion

Conclusions:

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

No increase of the number of histidine-prototrophic mutants after treatment with the test substance either with or without metabolic activation was observed, therefore, the test article is considered to be not mutagenic.

Executive summary:

The test substance was tested for mutagenic effects on histidine-auxotrophic mutants of Salmonella typhimurium. The investigations were performed on strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538 with the following concentrations of the trial substance without and with microsomal activation: 20, 80, 320, 1280 and 5120 µg/0.1 ml. In order to confirm the results, the experiments were repeated. In the experiments performed without and with microsomal activation, comparison of the number of back-mutant colonies in the controls and the cultures treated with the various concentrations of the test article revealed no marked deviations. No evidence of the induction of point mutations by the test substance or by the metabolites of the substance formed as a result of microsomal activation was detectable in the strains of S. typhimurium used in these experiments.