Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: not stricty according to OECD 471, no statistical analysis
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1987
Report date:
1987

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
not stricty according to OECD 471, no statistical analysis
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1,3,5-tris(oxiranylmethyl)-1,3,5-triazine-2,4,6(1H,3H,5H)-trione
EC Number:
219-514-3
EC Name:
1,3,5-tris(oxiranylmethyl)-1,3,5-triazine-2,4,6(1H,3H,5H)-trione
Cas Number:
2451-62-9
Molecular formula:
C12H15N3O6
IUPAC Name:
tris[(oxiran-2-yl)methyl]-1,3,5-triazinane-2,4,6-trione
Details on test material:
Araldite PT810 (TGIC), 1,3,5-triglycidylisocyanurate.
Purity: Commercial grade (> 97%)
Impurity < 100 ppm epichlorohydrin (CAS-no. 106-89-8)

Method

Target gene:
histidine auxotrophe
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
S. typhimurium TA 1538
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9- rat liver extract Arochlor 1254 induced
Test concentrations with justification for top dose:
1.22 - 10000 microgram/plate
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
Migrated to IUCLID6: 2-nitrofluorene, quinacrine mustard, 2-anthramine
Details on test system and experimental conditions:
Positive control substances were Sodium azide (in water) for nonactivated TA-1535 & TA-100; 2-nitrofluorene (in DMSO) for nonactivated TA-1538 & TA-98; Quinacrine mustard (in DMSO) for nonactivated TA-1537; 2-Anthramine (in DMSO) for all activated strains
All strains are histine auxotrophs; only revertant mutations to histidine independence are growing in medium with minimal amounts of histidine. Spontaneous reverse mutations are rare, and only mutation rates of 2-100 fold are considered positive. Strains were received from Bruce Ames.
Media used were: Oxoid Media #2 (nutrient broth) for overnight growth at 37°C; Selection media for mutagenicity assay was Vogel Bonner Medium E (Vogen and Bonner, 1956) with 2% glucose and 1.5% bactoagar.; overlay medium was 0.6% purified agar, 10% of 0.5 mM L-Histidine and 0.5 mM Biotin, and 0.5% NaCl according to Ames et al (1975).
Assay were performed with and without S9 activating homogenate.
S9 was 9000 xg supernatant from Sprague-Dawley adult male rat liver induced by Aroclor 1254. The S9 mix consited of NADP (sadium salt) 4 μmoles, D-Glucose-6-phosphate 5 μmoles, MgCl2 8 μmoles, KCl 33 μmoles, Na-Phosphate buffer pH 7.4 100 μmoles, and S9 homogenate 100 μlitres
Dose selection was performed with TA-100 in the non-activated system in overlay agar contailing TGIC (0.05 -0.1 ml of appropriate solution), 0.1-0.2 ml indicator organisms, and 0.5 ml 0.2M phosphate buffer pH 7.4. This solution (at 43-45°C) was added to minimal agar plates and incubated at 37°C for two days. Colonies were counted manually on the plates. Reduction of colony count was considered as toxicity.
Main assay conduct:
Non-activated: 2 ml Overlay agar, 0.05-0.1 ml TGIC solution , 0.2 ml indicator organism, and 0.5 ml 0.2M Phosphate buffer pH 7.4, was swirled gently and poured over a minimal agar plate and incubated for 48 hours. Colonies were counted manually on the plates.
Activated: Same as non-activated , but instead of 0.5 ml =.2M Phosphate buffer pH 7.4, 0.5 ml of S9 mix is added.
Negative controls were run in parallel (only overlay agar with test organisms).
Strains TA-98 and TA-100: at least three concentrations in a row have to be positive, and the response has to be at least 2-times the control value.
Control values were : TA-1535: 8-30; TA-1537: 4-30; TA-1538: 10-40; TA-98: 20-75; TA-100: 80-250.
Experiments were performed in triplicates, and 2 independent experiments were performed.
Evaluation criteria:
Evaluation of data: Strains TA-1535, TA-1537, TA-1538: at least three concentrations in a row have to be positive, and the response has to be at least 3-times the control value.
Statistics:
No statistical analysis of the data was performed.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
ambiguous
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

The assay was positive (mutagenic activity associated with TGIC, araldite PT 810).

Doses assayed were from 1.22 to 10’000 μg/plate, showing some toxicity 5000 and complete toxicity at 10’000 μg/plate with TA-100 in the preliminary assay.

Therefore, doses of 1-10’000 μg/plate were used throughout the experiment with all strains.

The positive controls were within the expected range of induced colonies

Applicant's summary and conclusion

Conclusions:
TGIC caused mutagenic effects in all strains tested with the exception of TA 1538 (ambiguous results) in the Ames test, and thus, it is mutagenic in-vitro
Executive summary:

Araldite PT 810 (TGIC) was mutagenic in three different Salmonella strains (TA-1535, TA-98 and TA-100) in the presence and absence of S9-activating enzymes of rat liver induced with Aroclor 1254.Tehrefore, Araldite PT 810 is mutagenic in bacteria in-vitro. Based on the results of the non-activating system, it is obvious that TGIC is  a Direct-Acting Mutagen