Pentasodium bis[5-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-4-hydroxy-3-[(2-hydroxy-5-nitrophenyl)azo]naphthalene-2,7-disulphonato(4-)]chromate(5-)

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996-1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

other: EEC Directive 92/69, L383, Annex V, B12, dated December 29, 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

FAT 40210/E

IUPAC Name:

FAT 40210/E

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

purity approx. 90%, red-violet solid

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animals: Strain-NMRI; Source: BRL, CH-4414 Füllinsdorf;
Number of Animals: 36 males/36 females;
Initial Age at start of Acclimatization: 8-12 weeks;
Acclimatization: minimum 5 days;
Initial body weight at start of treatment: males mean value 31.0g (SD±2.5g), female mean value 23.4 g (SD±1.5g).
The animals were in healthy condition and under quarantine in the animal house of CCR for a minimum of five days after their arrival. During this period the animals did not show any signs of illness or altered behavior. The animals were distributed into the test groups at random and identified by cage number.

Environmental conditions
Husbandry: the animals were kept conventionally. The experiment was conducted under standard laboratory conditions;
Housing: single; Cage Type: Makrolon Type I, with wire mesh top (EHRET GmbH, D-79302 Emmendingen);
Bedding: granulated soft wood bedding (ALTRONIM, D-32791 Lage/Lippe);
Feed: pelleted standard diet, ad libitum ((ALTRONIM 1324, D-32791 Lage/Lippe);
Water: tap water, ad libitum, (Gemeindewerke, D-64380 Robdarf); E
nvironment: temperature 21±3 ℃, relative humidity 30-70 %, artificial light 6.00 a.m.-6.00 p.m.

Administration / exposure

Route of administration:

oral: unspecified

Vehicle:

deionised water

Details on exposure:

test material: a single standard volume of 20ml/kg bogy weight orally (200, 670 and 2000 mg/kg b.w. respectively);
Vehicle control: orally, once, 20 ml/kg b.w.;
Positive control: orally, once, 40 mg/kg b.w.(20 mg/kg b.w.)

Duration of treatment / exposure:

24 h and 48 h

Frequency of treatment:

once

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

200 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

670 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

2 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5 per sex per dose

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide

Examinations

Tissues and cell types examined:

polychromatic erythrocytes

Details of tissue and slide preparation:

The animals were sacrificed by cervical dislocation. The femora were removed, the epiphyses were cut off and the marrow was flushed out with fetal calf serum, using a syringe. The cell suspension was centrifuged at 1500 rpm (390 X g) for 10 minutes and the supernatant was discarded. A small drop of the resuspended cell pellet was spread on a slide. The smear was air-dried and then stained with May-Grunwald (MERCK, D-64293 Darmstadt)/Giemsa (Gurr, BDH Limited Poole, Great Britain). Cover slips were mounted with EUKITT (KINDLER, D-79110 Freibur). At lease one slide was made from each bone marrow sample.

Evaluation criteria:

A test article is classified as mutagenic if it induces either a dose-related increase in the number of micronucleated polychromatic erythrocytes or a statistically significant positive response for at least one of the test points. A test article producing neither a dose-related increase in the number of micronucleated polychromatic erythrocytes nor a statistically significant positive response at any of the test points is considered non-mutagenic in this system. This can be confirmed by means of the nonparametric Mann-Whitney test. However, both biological and statistical significance should be considered together.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Remarks:

did not induce micronuclei as determined by the micronucleus test in the bone marrow cells of the mouse

Toxicity:

no effects

Remarks:

None of the animals expressed toxic reactions treated with 2000 mg/kg b.w. and FAT 40210/E had no cytotoxic properties in the bone marrow.

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

FAT 40210/E was considered to be non-mutagenic in the micronucleus assay.

Executive summary:

The study was performed to investigate the potential of FAT 40210/E to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse.

The following dose levels of the test article were investigated: 24 h preparation interval: 200, 670 and 2000 mg/kg b.w.(20ml/kg b.w.), 48 h preparation interval: 2000 mg/kg b.w.(20 ml/kg b.w.).

FAT40210/E had no cytotoxic effectiveness in the bone marrow and did not induced micronuclei by the micronucleus test with bone marrow cells of the mouse. FAT40210 was considered to be non-mutagenic in the micronucleus assay according to CLP Regulation (Regulation EC No. 1272/2008).