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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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-report according to guideline.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
from August 1983
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): n-butylchloride
- Storage condition of test material: cool, well ventilated place

Test animals

Species:
mouse
Strain:
other: BOR:NMRI
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Firma Winkelmann, Versuchstierzucht, Borchen, Germany
- Age at study initiation: about 3 months
- Weight at study initiation: males: 33.7 – 40.0 g, females: 29.0 – 35.5 g
- Assigned to test groups randomly: yes, under following basis: by lot
- Housing: collective caging (max. 5 animals per sex) in Macrolon type III cages
- Diet: ad libitum Ssniff-R Alleindiät für Ratten (Sniff Spezialitäten GmbH, Soest, Germany)
- Water: ad libitum aqua fontana; regularly analytical and bacteriological controls
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5 +/- 1.5
- Humidity (%): 55 +/-10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: April, 28 th 1989 To: May, 12th 1989

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) used: corn oil
- Amount of vehicle (if gavage or dermal): 10 mL/kg bw
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The test solution was prepared by suspending appropriate amounts with corn oil

Duration of treatment / exposure:
24, 48 or 72 hours
Frequency of treatment:
single dosage
Post exposure period:
no
Doses / concentrations
Remarks:
Doses / Concentrations:
2000 mg/kg bw in the main study
Basis:
no data
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide
- Route of administration: intraperitoneal
- Doses / concentrations: 40 mg/kg bw

Examinations

Tissues and cell types examined:
Bone marrow smears (at least two slides per animal) from the first 5 animals per sex and group were used for evaluation.
One slide per animal was examined (blind evaluation).
From each animal 1000 polychromatic erythrocytes (PCE) were scored for the incidence of micronuclei.
The ratio of polychromatic to normochromatic erythrocytes (PCE/NCE) was calculated on the base of 1000 cells.
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: preliminary dose-range-finding study

DETAILS OF SLIDE PREPARATION: After sacrification the femura were removed and the bond marrow was suspended in fetal calf serum. Samples were centrifuged at 1600 g, decanted and then one drop of each single suspension was smeared on a slide. These preparations were dried, fixed in absolute methanol and dried. The slides were stained using a May-Gruenwald and Giemsa solution.


Statistics:
One factorial analysis of variance; group mean values were compared by the method of Scheffé.
Significant levels: p > 0.05; p> 0.01; p > 0.001

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Remarks:
for details see results
Vehicle controls validity:
valid
Negative controls validity:
other: yes; historical controls
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 1000, 2500 and 5000 mg/kg bw (2 mice each)
- Clinical signs of toxicity in test animals: dosages of 2500 and 5000 mg/kg bw caused ataxia, writhing reflex, piloerection and a slight sedation over a period of 4 hours p.a.. Very slight findings of sedation and piloerection were seen at a dosage of 1000 mg/kg bw.

Any other information on results incl. tables

  PCE with micronuclei per 1000 PCE PCE per 1000 erythrocytes NCE per 1000 erythrocytes
solvent control  2.4 +/- 1.7 400.8 +/- 65.8 599.2 +/- 65.8
positive control 26.0 +/- 12.2 456.6 +/- 67.7 543.4 +/- 67.7
24 hours 2.6 +/- 1.1 396.4 +/- 81.1 603.6 +/- 81.1
48 hours 1.7 +/- 1.4 415.9 +/- 45.9 584.1 +/- 45.9
72 hours 1.0 +/- 0.7 474.1 +/- 73.0 525.9 +/- 73.0

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative