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Toxicological information

Genetic toxicity: in vitro

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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

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1987
Report Date:
1987
Reference Type:
publication
Title:
Unnamed
Year:
2002

Materials and methods

Test guideline
Qualifier:
according to
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

Reference
Name:
Unnamed
Type:
Constituent
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.
Controlsopen allclose all
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)
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 resultsopen allclose all
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
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
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
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)