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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
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (purity not reported)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report date:
1981

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
(1997)
GLP compliance:
no
Type of assay:
micronucleus assay

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N'-phenylene-1,4-bis[4-[(2,5-dichlorophenyl)azo]-3-hydroxynaphthalene-2-carboxamide]
EC Number:
223-460-6
EC Name:
N,N'-phenylene-1,4-bis[4-[(2,5-dichlorophenyl)azo]-3-hydroxynaphthalene-2-carboxamide]
Cas Number:
3905-19-9
Molecular formula:
C40H24Cl4N6O4
IUPAC Name:
3,3'-[(2,2'',5,5''-tetrachloro-1,1':4',1''-terphenyl-4,4''-diyl)bis(triaz-2-ene-3,1-diylcarbonyl)]di(2-naphthol)
Test material form:
not specified
Details on test material:
- Physical state: no data

Test animals

Species:
hamster, Chinese
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Weight at study initiation: male: 23-30 g, female: 21-29 g
- Housing: single cage (individually)
- Diet (e.g. ad libitum): NAFAG No. 924
- Water (e.g. ad libitum): ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24°C
- Humidity (%): 46 - 70%
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: PEG 400
- Concentration of test material in vehicle: 62.5 , 125, 250 mg/mL
- Amount of vehicle (if gavage or dermal): 20 mL/kg
Details on exposure:
after 48 hrs of first application animals were sacrificed
Duration of treatment / exposure:
48 hrs
Frequency of treatment:
2 doses per animal; once daily
Post exposure period:
24 hrs
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
1250 mg/kg bw
Basis:

Remarks:
Doses / Concentrations:
2500 mg/kg bw
Basis:

Remarks:
Doses / Concentrations:
5000 mg/kg bw
Basis:

No. of animals per sex per dose:
6
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide:
- Route of administration: oral
- Doses / concentrations: 128 mg/kg in 20 mL/kg PEG 400

Examinations

Tissues and cell types examined:
Blood cells of bone marrow
Details of tissue and slide preparation:
Bone marrow was harvested from the shafts of both femurs. In a siliconized pipette filled with approx. 0.5 μL rat serum the bone marrow was drawn up. Small drops of the mixture were transferred on the end of a slide, and the preparations were air-dried. After 3 hrs, the slides were stained in undiluted May-Gruenwald solution for 2 min then in May-Gruenwald solution/water 1/1 for 2 min and then in Giemsa's, 40% for 20 min. After being rinsed in methanol 55% for 5-8 s and washed off twice in water, they were left immersed in water for approx. 2 min. After rinsing with distilled water and air-drying, the slides were cleared in Xylol and mounted in Eukitt.
Evaluation criteria:
1000 bone marrow cells each were scored per animal and the following anomalies were registered: a) Single Jolly bodies, b) fragments of nuclei in erythrocytes, c) micronuclei in erythroblasts, d) micronuclei in leucopoietic cells, e) polyploid cells.
Statistics:
Significance of differences between doses were assessed by chi-square test

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
One female animal of the low-dose group died after the 1st application. Due to lack of dose-dependency and affection of a single animal, a treatment-related but not a substance-related effect is assumed.

Any other information on results incl. tables

In all treatments with the substance and in the negative control, the mean percentage of anormal cells was 0.1 while the positive control substance caused 10% abnormal cells, indicating the absence of a genotoxic effect in this test system and a suitable sensitivity of the test system (see table).

  dose
 [mg/kg bw]
average number
of cell anomalies
per animal
negative control
(PEG 400)
  0,1
positive control
(Cyclophosphamide)
128 10
test substance 1250 0,1
test substance 2500 0,1
test substance 5000 0,1

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative