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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 March 1983
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1983
Report date:
1983

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: EEC Directive 79/831, Annex V, Part B, Paragraph 4.3.1
Deviations:
no
GLP compliance:
no
Type of assay:
mammalian erythrocyte micronucleus test

Test material

Constituent 1
Reference substance name:
Disodium 1-amino-4-(4-benzenesulfonamido-3-sulfonatoanilino)anthraquinone-2-sulfonate
EC Number:
400-350-8
EC Name:
Disodium 1-amino-4-(4-benzenesulfonamido-3-sulfonatoanilino)anthraquinone-2-sulfonate
Cas Number:
85153-93-4
Molecular formula:
C26H17N3Na2O10S3
IUPAC Name:
disodium 1-amino-4-(4-benzenesulfonamido-3-sulfonatoanilino)anthraquinone-2-sulfonate
Specific details on test material used for the study:
Name: FAT 20297/C
Purity: 98 %

Test animals

Species:
mouse
Strain:
other: OF-1 albino
Details on species / strain selection:
Recommended by the Guideline
Sex:
male/female
Details on test animals or test system and environmental conditions:
The experiment was performed using non-consanguinous albino mice originating from an SPF colony (IFFA-CREDO, L'Arbresle, France). Mice of both sexes (25 g) were used after a quarantine period of 2 weeks at Battelle; during the quarantine the animals were allowed ad libitum access to food (Aliment Rats-Souris Charles River, produced by U.A.R., Villemoisson, France) and drinking water. Animals are housed 5 of the same sex by cage in Makrolon type III cages.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
The test substance was dissolved in distilled water.
Duration of treatment / exposure:
1. Control (distilled water): 0.5 mL - sacrificed at 20 hours
2. Positive control (Thio-TEPA): 80 mg/kg bw - sacrificed at 20 hours
3. FAT 20297/C: 1500 mg/kg bw - sacrificed at 20 hours
4. FAT 20297/C: 1500 mg/kg bw - sacrificed at 44 hours
5. FAT 20297/C: 1500 mg/kg bw - sacrificed at 68 hours.
Frequency of treatment:
Once
Doses / concentrations
Dose / conc.:
1 500 other: mg/kg bw
No. of animals per sex per dose:
5
Control animals:
yes
Positive control(s):
Thio-TEPA (N,N', N"-triethylenethiophosphoramide)
- Route of administration: oral
- Doses: 80 mg/kg bw

Examinations

Tissues and cell types examined:
Bone marrow and micronucleated polychromatic erythrocytes
Details of tissue and slide preparation:
After sacrifice of the animals the femurs were taken and broken open at one end. Bone marrow cells were suspended in foetal calf serum using a small syringe and the cells were centrifuged at 120 x g for 5 minutes. The supernatant was removed with a Pasteur pipette, cells were spread on a microscope slide and the smears allowed to dry in air. The following day smears were stained with Giemsa (1:6 in water) and mounted with a coverslip after drying.
Evaluation criteria:
Statistically significant increase in the number of micronucleated polychromatic erythrocytes
Statistics:
using BMPD computer programme 7D

Results and discussion

Test results
Key result
Sex:
male/female
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

GENERAL APPEARANCE OF THE SMEARS:

At low magnification of the microscope no noticeable differences in bone marrow nucleated cells were observed between animals treated with FAT 20297/C and negative control.

In the positive control group (Thio-TEPA) decreased numbers of bone marrow nucleated cells were noted.

NUMBERS OF OBSERVED MICRONUCLEI AND RATIO OF POLYCHROMATIC TO NORMOCHROMATIC ERYTHROCYTES

There was no statistical difference or sex effect between animals exposed to FAT 20297/C and negative control animals. There was no increase in the number of micronucleated polychromatic erythrocytes in animals exposed to 1500 mg/kg bw of FAT 20297/C. In animals treated with Thio-TEPA there is a statistically significant increase number of micronucleated cells.

The ratio of polychromatic to normochromatic erythrocytes in both control and FAT 20297/C treated animals is the same. This ratio is decreased in mice treated with Thio-TEPA.

Applicant's summary and conclusion

Conclusions:
FAT 20297/C was considered to be non-clastogenic in the micronucleus assay.
Executive summary:

FAT 20297/C was assayed for mutagenicity using the micronucleus test. This test was conducted in accordance with EEC Directive 79/831, Annex V, Part B, Paragraph 4.3.1.

The compound was administered orally to mice at a concentration of 1500 mg/kg body weight. No increase in the number of micronucleated polychromatic erythrocytes was observed in the bone marrow smears taken 20, 44 and 68 hours after administration of the test substance. A positive control (Thio-TEPA) administered at a concentration of 80 mg/kg body weight showed pronounced evidence of mutagenicity 20 hours after administration, proving the sensitivity of the testing protocol.

Hence, FAT 20297/C was considered to be non clastogenic in this micronucleus assay.