1,3,5-triazine-2,4,6(1H,3H,5H)-trione, compound with 1,3,5-triazine-2,4,6-triamine (1:1)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Adquately documented publication on cyanurate.

Data source

Reference

Reference Type:

publication

Title:

Absence of mutagenic activity for monosodium cyanurate

Author:

Hammond, BG, Barbee, SJ, Wheeler AG and Cascieri, T

Year:

1985

Bibliographic source:

Fundamental and Applied Toxicology 5, pp. 655-664

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

yes

GLP compliance:

not specified

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2624-17-1

Cas Number:

2624-17-1

IUPAC Name:

2624-17-1

Constituent 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

water

Test material form:

solid: crystalline

Details on test material:

- name in publication: monosodium cyanurate
- supplier: Monsanto company
- composition: 77% cyanurate, 13% sodium and 10% water
- solubility in water: 0.7% w/v

Specific details on test material used for the study:

- name in publication: monosodium cyanurate
- supplier: Monsanto company
- composition: 77% cyanurate, 13% sodium and 10% water
- solubility in water: 0.7% w/v

Method

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Metabolic activation system:

liver S9 preparted from arochlor 1254-induced Sprague-Dawley rats

Test concentrations with justification for top dose:

50 - 2000 μg/plate (toxicity test)
250, 500, 750, 1000, 1250, 1500, 1750 and 2000 μg/plate (mutagenicity test)

Vehicle / solvent:

cell culture medium (F10P) or S-9 mix

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

Remarks:

with S9

Controls 2

Positive control substance:

ethylmethanesulphonate

Remarks:

without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk


DURATION
- Preincubation period: no data
- Exposure duration: 4h
- Expression time (cells in growth medium): 48 h
- Selection time (if incubation with a selection agent): 12 days
- Fixation time (start of exposure up to fixation or harvest of cells):


SELECTION AGENT (mutation assays): trifluorothymidine



NUMBER OF REPLICATIONS: triplicate


NUMBER OF CELLS EVALUATED: Clones growing from 200 cells/plate


DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency

Evaluation criteria:

Statistic difference to vehicle control

Statistics:

not required

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Any other information on results incl. tables

Relative cloning efficiencies ranged from 86 - 111% and 88 - 106% for all incubations with and without metabolic activation, respectively.

	Mutant frequency (10exp-6)	Mutant frequency (10exp-6)
	with S9	without S9
Solvent control	47	42
positive control	538	929
2000	48	58±9
1750	63±17	23±6
1500	39±18	46±9
1250	53±5	41
1000	52±5	40±2
750	48±11	culture lost
500	37±2	36±5
250	58±3	35±5

Applicant's summary and conclusion

Conclusions:

Sodium cyanurate is not mutagenic in mammalian cells in vitro.