Tris(2-hydroxyethyl)-1,3,5-triazinetrione

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented and reported study fully adequate for assessment, although not GLP compliant. The study was conducted according to an internationally accepted technical guideline in a recognized contract research organization.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Reference 2

Reference Type:

publication

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

of 1983

Deviations:

no

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Tris(2-hydroxyethyl)-1,3,5-triazinetrione

EC Number:

212-660-9

EC Name:

Tris(2-hydroxyethyl)-1,3,5-triazinetrione

Cas Number:

839-90-7

Molecular formula:

C9H15N3O6

IUPAC Name:

tris(2-hydroxyethyl)-1,3,5-triazinane-2,4,6-trione

Details on test material:

- Name of test material (as cited in study report): Trishydroxyethylisocyanurat (THEIC)
- Test-substance No.: 87/556
- Degree of purity: ca. 98 %
- Storage: +4°C

Method

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: essential amino acid requiring strains

Metabolic activation:

with and without

Metabolic activation system:

liver S9 mix from male Sprague Dawley rats treated by single intraperitoneal injection with Aroclor 1254 (500 mg/kg bw) for enzyme induction. Aroclor 1254 was administered 5 days prior to sacrifice.

Test concentrations with justification for top dose:

Experiment 1 (Standard Plate Test, without and with metabolic activation (S9 mix), Doses: 0; 20; 100; 500; 2500 and 5000 μg/plate
Experiment 2 (Pre-incubation Test, without and with metabolic activation (S9 mix), Doses: 0; 20; 100; 500; 2500 and 5000 μg/plate

Vehicle / solvent:

aqua dest.

Controls

Controls 1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

without and with S9 mix

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (disolved in DMSO)

Remarks:

Positive control substance for all tests with metabolic activation (S9 mix)

Controls 2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

without and with S9 mix

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine for TA 1535 & TA 100; 4-nitro-o-phenylendiamine for TA 98; 9-aminoacridine chloride monohydrate for TA 1537 (each positive control substance disolved in DMSO)

Remarks:

Positive control substances for tests without metabolic activation (S9 mix). All of them are well established reference mutagens.

Details on test system and experimental conditions:

Standard Plate Tests were performed in Experiment 1, Pre-incubation Tests in Experiment 2.
Both experiments were conducted without and with metabolic activation (S9 mix)

The following positive controls were used to check mutability of the bacteria and activity of the S9 mix:

With metabolic activation (S9 mix):

2-aminoanthracene:
- 10 μg/plate, dissolved in DMSO: for the strains: TA 1535, TA 100, TA 1537, TA 98

Without metabolic activation (S9 mix):

N-methyl-N'-nitro-N-nitrosoguanidine:
- 5 μg/plate, dissolved in DMSO: for the strains: TA 1535, TA 100

4-nitro-o-phenylendiamine:
- 10 μg/plate, dissolved in DMSO: for the strain: TA 98

9-aminoacridine chloride monohydrate:
- 100 μg/plate, dissolved in DMSO: for the strain: TA 1537

Evaluation criteria:

In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control),
- dose-response relationship,
- reproducibility of the results.

Statistics:

Statistical analysis of the results was not reported. Obviously it was not considered to be necessary.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

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:

The test substance was completely soluble in the vehicle, aqua dest.

Contamination was noted only in one plate which was one of the TA 100 without metabolic activation at 20 µg/plate. In all other solvent control, test substance treated or positive control plates contamination was not evident.

Applicant's summary and conclusion

Conclusions:

Interpretation of results :
negative without and with metabolic activation (S9 mix)