1,3,5-tris(3,5-di-tert-butyl-4-hydroxybenzyl)-1,3,5-triazine-2,4,6(1H,3H,5H)-trione

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

Study period:

1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guideline study performed under GLP-like quality control conditions with QAU statement.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no statisical analysis

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,3,5-tris(3,5-di-tert-butyl-4-hydroxybenzyl)-1,3,5-triazine-2,4,6(1H,3H,5H)-trione

EC Number:

248-597-9

EC Name:

1,3,5-tris(3,5-di-tert-butyl-4-hydroxybenzyl)-1,3,5-triazine-2,4,6(1H,3H,5H)-trione

Cas Number:

27676-62-6

Molecular formula:

C48H69N3O6

IUPAC Name:

1,3,5-tris(3,5-di-tert-butyl-4-hydroxybenzyl)-1,3,5-triazine-2,4,6(1H,3H,5H)-trione

Details on test material:

- Physical state: solid
- Analytical purity: commercial grade

Method

Target gene:

histidine

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction of rat liver induced with Aroclor 1254

Test concentrations with justification for top dose:

0.08 - 5000 µg/0.1 ml, in the toxicity test
20, 78, 313, 1250 and 5000 ug/0.l ml, in the mutagenicity test

Vehicle / solvent:

- Vehicle used: acetone

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: TA 98: daunorubicin- HCl; TA 100: 4-nitroquinoline-N-oxide; TA 102: mitomycin-C; TA 1535: sodium azide; TA1537: 9(5)-aminoacridine hydrochloride monohydrate

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: TA 98, TA 100, TA 102, TA 1537: 2 - aminoanthracene; TA 1535: cyclophosphamide

Remarks:

with metabolic activation

Details on test system and experimental conditions:

Each Petri dish contained: approx. 20 ml of minimum agar (plus salts and glucose), 0.1 ml of a solution of the test substance or the vehicle and 0.1 ml of a bacterial culture in 2.0 ml of soft agar. In the experiments in which the substance was metabolically activated, 0.5 ml of an activation mixture was added also. 1 ml activation mixture contained: 0.3 ml S9 fraction of liver from rats (Tif:RAIf(SPF)) induced with Aroclor 1254 and 0.7 ml of a solution of co-factors.
In the experiments without and with the addition of microsomal activation mixture three Petri dishes were prepared per strain and per group (i.e. per concentration or per control group). The plates were incubated for about 48 hours at 37 +/- 1.5 °C in darkness.
The experiment was repeated for confirmation.

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:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

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:

At the concentrations of 1250 µg/0.1 ml and above 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

Without S9 -Mix

Concentration µg/0.1 ml	TA 98		TA 100		TA 102		TA 1535		TA 1537
	I	II	I	II	I	II	I	II	I	II
Solvent Control	26	37	238	160	362	322	18	17	7	7
Positive Control	873	992	959	1123	1163	1221	1289	1364	1069	1287
20	22	25	241	140	362	311	17	17	6	7
78	25	28	220	152	347	329	18	15	11	7
313	24	26	212	133	347	282	13	16	11	7
1250	23	26	176	158	315	217	16	16	7	7
5000	30	18	185	111	354	245	13	13	7	3

I = experiment 1

II = experiment 2

Positive controls:

daunorubicin-HCl (10 µg/0.1 ml) for TA 98;

4 -nitroguinoline-N-oxide (0,25 µg/0.1 ml) for TA 100;

mitomycin-C (1 µg/0.1 ml) for TA 102

sodium azide (5.0 µg/0.1 ml) for TA 1535

9 (5) aminoacridine hydrochloride (100 µg/0.1 ml) for TA 1537

With S9 -Mix

Concentration µg/0.1 ml	TA 98		TA 100		TA 102		TA 1535		TA 1537
	I	II	I	II	I	II	I	II	I	II
Solvent Control	47	37	137	135	359	334	17	20	15	13
Positive Control	1098	1048	1401	1623	1413	1424	330	547	119	197
20	40	44	133	115	339	348	15	11	14	16
78	46	56	138	113	271	253	8	15	13	12
313	48	39	126	115	322	237	14	16	11	9
1250	34	39	127	110	258	286	15	16	15	7
5000	37	31	121	109	329	256	15	10	9	5

I = experiment 1

II = experiment 2

Positive controls:

2 -aminoanthracene (5 µg/0.1 ml) for TA 98, TA 100 and TA 1537;

2 -aminoanthracene (20 µg/0.1 ml) for TA 102;

cyclophosphamide (250 µg/0.1 ml) for TA 1537

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

During the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

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 102, TA 1535 and TA 1537 with the following concentrations of the trial substance without and with microsomal activation: 20, 78, 313, 1250 and 5000 µg/0.l ml. In order to confirm the results the experiments were repeated. Any mutagenic effects of the substances are demonstrable on comparison of the number of bacteria in the treated and control cultures that have undergone back-mutation to histidine-prototrophism. In the experiments performed without and with microsomal activation, comparison of the number of back-mutants in the controls and the cultures treated with the various concentrations of the test substance 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.