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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / micronucleus study
Remarks:
Type of genotoxicity: genome mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-06-01 to 2012-12-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study, detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: OECD 487
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
in vitro mammalian cell micronucleus test

Test material

Constituent 1
Chemical structure
Reference substance name:
6,6',6''-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoic acid
EC Number:
279-505-5
EC Name:
6,6',6''-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoic acid
Cas Number:
80584-91-4
Molecular formula:
C21H36N6O6
IUPAC Name:
6,6',6''-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoic acid
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): TC, dried
- Physical state: White powder
- Analytical purity: 95.57%
- Moisture: 0.05%
- Melting point: 181-185 °C
- Lot/batch No.: 110525S
- Expiration date of the lot/batch: 25-05-2016
- Storage condition of test material: ambient temperature (15-25°C)
- Other: soluble in DMSO

Method

Target gene:
Formation of micronuclei in human peripheral blood lymphocytes
Species / strain
Species / strain / cell type:
lymphocytes:
Metabolic activation:
with and without
Metabolic activation system:
Mammalian liver post-mitochondrial fraction (S9), prepared from Sprague Dawley male rats
Test concentrations with justification for top dose:
Range-finding: 0, 9.375, 18.75, 37.5, 75, 150, 300 µg/mL (final concentrations)
Definitive test. experiment 1, -S9: 75, 150, 300 µg/mL + vehicle (1% DMSO v/v ) + Cytosine arabinoside (pos. control)
Definitive test. experiment 1, +S9: 75, 150, 300 µg/mL + vehicle (1% DMSO v/v ) + Cytosine arabinoside (pos. control)
Definitive test. experiment 2, -S9: 7.5, 15, 30, 60, 120, 240, 480 µg/mL + vehicle (1% DMSO v/v ) + DMBA (pos. control)
Definitive test. experiment 2, +S9: 7.5, 15, 30, 60, 120, 240, 480 µg/mL + vehicle (1% DMSO v/v ) + DMBA (pos. control)
Appropriate amount of test item was weighted and diluted in DMSO not earlier than 30 min prior to test start. For dilution of final doses, sterile DMSO was used.
Vehicle / solvent:
- Vehicle/solvent used: DMSO+ culture medium
Controls
Untreated negative controls:
yes
Remarks:
culture medium
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
Cytosine arabinoside was used as positive control without S9mix, Cyclophosphamide monohydrate was used as positive control with S9mix.
Details on test system and experimental conditions:
METHOD OF APPLICATION:
10exp5 cells in 75 µl

DURATION
- Exposure duration: 20 hr

METHOD OF APPLICATION: in medium;

DURATION
- Preincubation period: 24 hours
- Exposure duration: 20 hours
- Time with Cytochalasin B: 28 hours:
- Fixation time (start of exposure up to fixation or harvest of cells): 72 hours

SPINDLE INHIBITOR (cytogenetic assays): Cytochalasin B
STAIN (for cytogenetic assays): 2% Giemsa-Romanowski for 10 minutes

NUMBER OF CELLS EVALUATED: 10exp5 cells

DETERMINATION OF CYTOTOXICITY
- Method: other: replication index (Number of binucleated cells, mulinucleated cells and toatal number)
Evaluation criteria:
For the untreated controls and solvent controls, the measured average number of micronuclei is related to the spontanous occurence of micronuclei. A two tailed Student's t-test revealed if the test substance can be considered as positive, negative or equivocal.
Statistics:
Student's t-test

Results and discussion

Test results
Species / strain:
lymphocytes: human
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

Any other information on results incl. tables

Experiment 1: range-finding test without metabolic activation

Experiment 2:definitive test with and without metabolic activation

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

TC, dried did not induce micronuclei neither with nor without metabolic activation by S-9 mix in human lymphocytes. TC, dried was considered to be non-mutagenic.
Executive summary:

TC, dried was dissolved in DMSO in concentrations of 9.375 to 300 µg/ml in an in vitro micronucleus test with human lymphocytes according to OECD 487.

Up to and including the highest possible concentrations TC, dried, did not induce micronuclei neither with nor without metabolic activation by S-9 mix.

Therefore TC, dried was considered to be non-mutagenic.

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