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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
Study period:
1981-06-02 to 1981-08-04
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions ( medullar toxicity is not checked through PCE/NCE ratio but through haematological analysis, 5 animals per sex are usually required (instead of only 4 here))

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)
Deviations:
yes
Remarks:
4 instead of 5 animals per sex
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Constituent 1
Reference substance name:
No IUPAC name
EC Number:
614-637-2
Cas Number:
68603-75-8
Molecular formula:
R-N(CH2CHOHCH3)xCH2CH2CH2N(CH2CHOHCH3)y with x+y=3 and where R = alkyl
IUPAC Name:
No IUPAC name
Details on test material:
- Name of test material (as cited in study report): DINORAM SL
- Physical state: dark brown liquid
- Lot/batch No.: 2884
No other data available

Test animals

Species:
mouse
Strain:
CD-1
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Source: Charles River France (76410 Saint-Aubin-lès-Elbeuf, France)
- Age: 13-17 weeks
- Body weight at study initiation: no data

Administration / exposure

Route of administration:
other: test substance gavage, positive controls intraperitoneal
Vehicle:
Vehicle: CMC at 1%
Duration of treatment / exposure:
2 administrations separated by 24h
Frequency of treatment:
2 administrations separated by 24h
Doses / concentrations
Remarks:
Doses / Concentrations:
0.1, 0.2 and 0.5 ml/kg
Basis:

No. of animals per sex per dose:
4 males + 4 females
Control animals:
yes, concurrent vehicle
Positive control(s):
Benzo(a)pyrene (500 mg/kg/day) and Cyclophosphamide (100 mg/kg/day) in corn oil

Examinations

Details of tissue and slide preparation:
- Haematology: blood samples were withdrawn from retro-orbital sinus for measuring red blood count, mean cell volume, hematocrit, hemoglobin, mean cell hemoglobin concentration and white cell count
- Tissue examined: blood and bone marrow - Actual dose (mg/kg body weight): not determined
- Slide preparation: At the scheduled sacrifice times (30 hours after the second administration for test substance and negative control groups, 48h hours after the second administration for positive control groups), mice were sacrificed by vertebras dislocation. Immediately following sacrifice, the femurs were removed and the bone marrow was flushed with fetal calf serum. The bone marrow cells were transferred to labelled centrifuge tubes. The bone marrow cells were pelleted by centrifugation at 1200 g for 5 minutes and the supernatant was drawn off, leaving a small amount of serum with the remaining cell pellet. The cells were resuspended with serum and a small volume of bone marrow suspension was spread onto two clean glass slides. The slides were stained with May-Grünwald-Giemsa.
- PCE/NCE ratio was determined until a total of at least 1000 cells (PCE+NCE) were counted Each slide is examined by two persons.
- A total of 1000 PCE was examined for micronuclei.
Statistics:
Comparisons between treated and control groups were performed using a Student Fischer test.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
- Mortality: No death occured in test substance group but a male was found dead after the second administration.
- Clinical findings: No pathological effects were reported.
- Haematology: No significant differences in haematological parameters between treated and negative control groups were noted. Notably, no anemia were recorded.
- Micronuclei count: No significant differences were observed between treated and negative control group. In contrast, positive control slides showed an increased micronuclei count.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
Under these experimental conditions, the substance did not show a clastogenic potential.
Executive summary:

Four male and four female mice were treated with 0.1, 0.2 or 0.5 mg/kg/d of Amines, N-tallow alkyltrimethylenedi-, propoxylated on two days. Blood of the animals was taken for hematological observations, bone marrow samples were taken for investigation of micronucleus formation. Neither in the blood samples nor in the bone marrow samples, differences between control animals and dosed animals were observed. The animals treated with positive controls (Benzo(a)pyrene and Cyclophosphamide), however did show a markedly increased number in micronuclei.