Registration Dossier

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:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
GLP compliance:
yes (incl. certificate)
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
mouse
Strain:
Swiss
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeder: Charles River Laboratories, l'Arbresle, France.
- Age at study initiation: approximately 6 weeks
- Weight at study initiation: males: 28 - 33 g; females 21 - 27 g
- Assigned to test groups randomly: yes, the animals were randomly allocated to the groups by sex
- Fasting period before study: no
- Housing: by groups in polycarbonate cages;
bedding: autoclaved sawdust (SICSA, Alfortville, France)
- Diet (ad libitum): Ssniff R/M-H pelleted maintenance diet (SSNIFF Spezialdiäten GmbH, Soest, Germany).
- Water (ad libitum): drinking water filtered by a FG Millipore membrane (0.22 micron)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C,
- Humidity (%): 30 - 70%
- Air changes (per hr): 12 cycles/hour of filtered non-recycled fresh air
- Photoperiod (hrs dark / hrs light): 12 h/12 h (07:00 - 19:00)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
Vehicle
Name: 0.5% methylcellulose

Positive controls
The positive control was Cyclophosphamide 50 mg/kg.
Details on exposure:
For the vehicle and the test item:
• Route: oral, since it is a possible route of exposure in Man
• Frequency: two treatments separated by 24 hours
• Volume: 10 mL/kg.

For the positive control (CPA):
• Route: oral
• Frequency: one treatment
• Volume: 10 mL/kg.
Duration of treatment / exposure:
48 hours (test item)
24 hours (cyclophosphamide)
Frequency of treatment:
two treatments separated by 24 hours
Post exposure period:
24 h
No. of animals per sex per dose:
Doses/Concentrations:
Test item: males: 250-500-1000 mg/kg/day
Females: 125-250-500 mg/kg/day

CPA: 10 mg/kg/day

No of animals per sex per dose
5 males and 5 females/doses including controls except 8 males and 8 females at the high dose
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide

Examinations

Tissues and cell types examined:
Femoral bone marrow.
Erythrocyte cell line.
Details of tissue and slide preparation:
Bone marrow flushed out with fetal colf serum. Centrifugation and cell suspension spread on the slide. Straining with Griensa.
Evaluation criteria:
-2000 polychromatic erythrocytes evaluated/animal for MPE.
-statisfically significant increase in MPE frequency versus controls
-historical date and biological relevance also considered
-PE/NE ratio: 1000 erythrocytes.
Statistics:
Yes.

Results and discussion

Test resultsopen allclose all
Sex:
male
Genotoxicity:
negative
Toxicity:
yes
Remarks:
One mortality at 1000 mg/kg/day. No effects on PE/NE.
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Sex:
female
Genotoxicity:
negative
Toxicity:
no effects
Remarks:
125, 250 and 500 mg/kg/day
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative