2,4,6-triallyloxy-1,3,5-triazine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-report according to guideline.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

2,4,6-triallyloxy-1,3,5-triazine

EC Number:

202-936-7

EC Name:

2,4,6-triallyloxy-1,3,5-triazine

Cas Number:

101-37-1

Molecular formula:

C12H15N3O3

IUPAC Name:

tris(prop-2-en-1-yloxy)-1,3,5-triazine

Method

Target gene:

not applicable

Species / strain

Species / strain / cell type:

lymphocytes: human, healthy donors without medication

Metabolic activation:

with and without

Metabolic activation system:

liver S9-mix of Aroclor 1254 induced rats

Test concentrations with justification for top dose:

0, 31.25, 62.5, 125, 250, 500 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was chosen according to its solubility properties and its non-toxicity for the cells.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO 1% (v/v)

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

without metabolic activation

Migrated to IUCLID6: 0.1 or 0.2 µg/mL

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO 1% (v/v)

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

with metabolic activation

Migrated to IUCLID6: 10 or 20 µg/ mL

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 48 h
- Exposure duration: 4 h or 24 h without metabolic activation, 4 h with metabolic activation
- Fixation time (start of exposure up to fixation or harvest of cells): 22 h



SPINDLE INHIBITOR (cytogenetic assays): colcemid 0.9 µg/mL
STAIN (for cytogenetic assays): Giemsa stain


NUMBER OF REPLICATIONS: two cultures each two slide preparations


NUMBER OF CELLS EVALUATED: 100 metaphase per slide were evaluated


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index


OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

Comparison of the number of chromosome aberrations of the samples with those of the solvent control. Total number of cells with aberrations exclusive of gap damage are analysed.
evaluation criteria:
The test is regarded to have a positive result, if the number of chromosomal aberrations is significantly increased (p<=0.05) compared with the solvent control, this increase is dose-dependent and both duplicate cultures lead to the same results. The increase should not occur in the severly cytotoxic range (mitotic index < 0.25).

Statistics:

Fisher-test (p<= 0.05)

Results and discussion

Test results

Species / strain:

lymphocytes: human, healthy donors without medication

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

125 µg/ mL 24 h exposure, 250 µg/ mL 4 h exposure

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: other: human peripheral lymphocytes

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Treatment	µg/mL	S9-mix	4 h exposure			24 h exposure
			Mitotic index#	number of metaphases scored	% of cells with aberrations excluding gaps	Mitotic index#	number of metaphases scored	% of cells with aberrations excluding gaps
DMSO	-	-	1.00	200	1.5	1.00	200	2.0
test substance	31.25	-	-	-	-	0.86	200	2.5
	62.5	-	0.92	200	1.5	0.55	200	3.5
	125	-	1.27	200	1.5	0.47	164#	3.7
	250	-	0.58	183#	4.5	0.28	35#	3.2
	500	-	0.43	101#	5.2	-	-	-
Mitomycin C	0.2	-	0.62	200	12.0*	0.55	200	13.0*

the following concentrations were not evaluated:

31.25 µg/ mL (4 h exposure), 500 µg/ mL ( 24 h exposure) 0.1 µg mitomycin C (4 and 24 h exposure)

Treatment	µg/mL	S9-mix	4 h exposure
			Mitotic index#	number of metaphases scored	% of cells with aberrations excluding gaps
DMSO	-	+	1.00	200	1.0
test substance	31.25	+	-	-	-
	62.5	+	0.77	200	2.0
	125	+	1.62	200	1.0
	250	+	0.40	183#	4.5
	500	+	0.13	11#	0.0
Cyclophosphamide	10	+	0.63	200	10.5*

the following concentrations were not evaluated:

31.25 µg/ mL, 20 µg/ mL cyclophosphamide

#          mitotic index: number of metaphases/ 1000 cells: negative control = 1.00
##        no more metaphase of sufficient quality for evaluation due to cytotoxicity of Triallylcyanurate
*          significantly different from negative control (p=0.05)

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative