Reaction mass of lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-[(4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]naphthalene-2,7-disulfonate and lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-{[4-(vinylsulfonyl)phenyl]diazenyl}naphthalene-2,7-disulfonate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Principles of method if other than guideline:

The test was conducted according to the method published by Salamone et al., 1980.

Salamone M, Heddle J, Stuart E and Katz M, 1980. Towards and improved micronucleus test. Studies on 3 model agents, mitomycin C, cyclophosphamide and dimethylbenzanthracene. Mutat. Res. 74:347-356.

GLP compliance:

yes

Type of assay:

other: Mammalian erythrocyte micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

Lithium sodium hydrogen 4-amino-6-(5-(5-chloro-2,6-difluoropyrimidin-4-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-2,7-disulfonate

EC Number:

401-560-2

EC Name:

Lithium sodium hydrogen 4-amino-6-(5-(5-chloro-2,6-difluoropyrimidin-4-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-2,7-disulfonate

Cas Number:

108624-00-6

Molecular formula:

C28H(21-x-y)ClF2Li(x)N8Na(y)O16S5

IUPAC Name:

Lithium sodium hydrogen-4-amino-6-(5-(5-chloro-2,6-difluoropyrimidine-4-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-2,7-disulfonate

Test material form:

solid: particulate/powder

Details on test material:

Reactive Blue FC 05717

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: F. Winkelmann, Borchen
- Age at study initiation: 8 to 12 wk
- Weight at study initiation: 23-34 g
- Housing: Makrolon Type II cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: one wk

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-22°C
- Humidity (%): 44-50%
- Photoperiod (hrs dark / hrs light): 12:12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: 0.5% aqueous cremophor emulsion
- Justification for choice of solvent/vehicle: the test substance has a better dispersibility in this vehicle
- Amount of vehicle (if gavage or dermal): 20 mL/kg bw

Duration of treatment / exposure:

24, 48 and 72 h for three separate test groups dosed at 7500 mg/kg bw
24 h for negative and positive control groups

Frequency of treatment:

Once

Doses / concentrations

Dose / conc.:

7 500 mg/kg bw (total dose)

No. of animals per sex per dose:

Ten (five males and five females) per dose group including negative and positive control groups

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide
- Route of administration: Oral gavage
- Doses / concentrations: 20 mg/kg bw
- Dose volume: 10 mL/kg bw

Examinations

Tissues and cell types examined:

Bone marrow from femur was obtained and the polychromatic erythrocytes were examined.

Details of tissue and slide preparation:

TREATMENT AND SAMPLING TIMES:
Three groups of mice were treated with the test substance and were sacrificed at 24 h interval (i.e., at 24, 48 and 72 h) to obtain the bone marrow samples.

DETAILS OF SLIDE PREPARATION:
The bone marrow preparation was stained with Ames Hema-Tek slide stainer, followed by treatment with methanol and washing with demineralised water. Once the slides are dried, they ere observed under microscope.

METHOD OF ANALYSIS: 1000 polychromatic erythrocytes were examined per animal and the frequency of micronucleated cells among them were determined.

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

- Following treatment with the test substance, no relevant increase in the number of micronucleated PCEs over the concurrent negative control was observed at the treated dose and at all the exposure intervals. A marked increase in the frequency of micronucleated PCEs was noticed in the positive control group.
- Bone marrow cell toxicity: not specified

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the test substance did not induce micronuclei in the polychromatic erythrocytes of the treated mice.

Executive summary:

A study was conducted to determine the in vivo genetic toxicity of the test substance in NMRI mice according to the method published by Solomone et al, 1980. The ability of the test substance to induce cytogenetic damage and/or disruption of the mitotic apparatus in rat bone marrow was investigated by measuring the induction of micronuclei in polychromatic erythrocytes. NMRI mice (5 per sex per group) were exposed to a single oral gavage dose of the test substance at 0 and 7500 mg/kg bw for 24, 48 and 72 h. A positive control group (cyclophosphamide, 20 mg/kg bw) was also tested. Following treatment with the test substance, no relevant increase in the number of micronucleated PCEs over the concurrent negative control was observed at any of the exposure interval. A marked increase in the frequency of micronucleated PCEs was noticed in the positive control group. The values for the positive and negative controls were within the expectation ranges. The experiment was therefore considered valid. Under the study conditions, the test substance did not induce micronuclei in the polychromatic erythrocytes of the treated mice (Herbold, 1986b).