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

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:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1977

Report date:

1977

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

No information on the purity of the substance and number of cells used. No individual plate counts, no statistical analyses. Only four strains tested.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

229-722-6

EC Name:

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

Cas Number:

6683-19-8

Molecular formula:

C73H108O12

IUPAC Name:

3-{[3-(3,5-di-tert-butyl-4-hydroxyphenyl)propanoyl]oxy}-2,2-bis({[3-(3,5-di-tert-butyl-4-hydroxyphenyl)propanoyl]oxy}methyl)propyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propanoate

Method

Target gene:

Histidine operon

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction of liver from rats induced with Aroclor 1254 plus co-factors

Test concentrations with justification for top dose:

without microsomal activation : 10, 25, 50, 100, and 250 μg/0.1 mL
with microsomal activation: 5, 10, 25, 50, and 100 μg/0.1 mL

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: see below

Remarks:

with and without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: 3 out of 6 plates per dose with activation were preincubated

NUMBER OF REPLICATIONS:
In the experiments without the addition of microsomal activation mixture, three Petri dishes were prepared per strain and per group (i.e. per concentration or per control group). In the experiments with activation mixture six Petri dishes were used per strain and per group; three of each set of six were pre-incubated

POSITIVE CONTROLS:
Without S9:
Strain TA 1535: N-methyl-N'-nitro-N-nitrosoguanidine, 2 µg/0.1 ml DMSO;
Strain TA 1537: 9(5)-aminoacridine hydrochloride monohydrate, 100 µg/0.1 ml DMSO
Strain TA 98: daunoblastin, 50 µg/0.1 ml DMSO;
Strain TA 100: 4-nitroquinoline-n-oxide, 10 µg/0.1 ml DMSO.
With S9:
Strain TA 100: cyclophosphamide, 100 and 250 µg/0.1 ml physiological saline

Evaluation criteria:

The test substance was considered to be non-mutagenic if the colony count in relation to the negative control was not doubled at any concentration.

Statistics:

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

Results and discussion

Test results

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:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: insoluble
- Precipitation: at 100 and 250 μg/0.1 mL 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

Table 1. Salmonella mutagenicity test: reverse mutation without microsomal activation. Numbers represent the arithmetic mean of histidine-prototrophic colonies.

Test substance	Strain of S. typhi-murium used
	TA 98	TA 100	TA 1535	TA 1537
control	28	115	11	8
10 μg/0.1 mL	24	118	11	8
25 μg/0.1 mL	24	151	14	5
50 μg/0.1 mL	24	149	17	6
100 μg/0.1 mL	24	136	13	7
250 μg/0.1 mL	24	163	16	7

Table 2. Salmonella/Mammalian microsome mutagenicity test: reverse mutation with microsomal activation. Numbers represent the arithmetic mean of histidine-prototrophic colonies.

n/n: first figures are values without, second figures values with pre-incubation.

Test substance	Strain of S. typhi-murium used
	TA 98	TA 100	TA 1535	TA 1537
control	18 / 21	173 / 165	18 / 20	8 / 7
5 μg/0.1 mL	23 / 25	179 / 202	17 / 17	9 / 9
10 μg/0.1 mL	22 / 25	181 / 188	21 / 20	8 / 5
25 μg/0.1 mL	23 / 21	190 / 217	26 / 20	7 / 8
50 μg/0.1 mL	23 / 23	177 / 189	21 / 19	7 / 7
100 μg/0.1 mL	23 / 22	157 / 155	17 / 18	7 / 7

Applicant's summary and conclusion

Conclusions:

In the experiments performed with and without microsomal activation, comparison of the number of histidine-prototrophic mutants in the controls and after treatment revealed no significant differences.