Registration Dossier

Toxicological information

Genetic toxicity: in vivo

Currently viewing:

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:
No data
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test).
Justification for data waiving:
other:
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report Date:
1985

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
not specified
Qualifier:
according to
Guideline:
other: EEC Directive 79/831, Annex V, Method No. 431
Deviations:
not specified
Principles of method if other than guideline:
Not applicable
GLP compliance:
not specified
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Ufanon K-80 (CAS 68603-42-9)
- Physical state: Brown viscous liquid
- Storage condition of test material: Room temperature

Test animals

Species:
mouse
Strain:
NMRI
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeders were purchased from G1. Bomholtgard Ltd., but the mice used were born in the Scantox Laboratories
- Age at study initiation: 6-7 wk
- Weight at study initiation: 25-29 g
- Assigned to test groups randomly: Yes, in 5 groups
- Fasting period before study: No
- Housing: 5 animals/cage, males and females separately in type III Macrolone cages, bedding used was special softwood sawdust "Spanvall Special White " from Spanvall Ltd., DK-4535 Vallekilde
- Diet: Complete rodent diet "Altromin 1314" from Chr. Petersen Ltd., DK-4100 Ringsted, ad libitum
- Water (e.g. ad libitum): Drinking water adjusted to pH 2.5 with hydrochloric acid


ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C
- Humidity (%): 55 ± 15%
- Air changes: 10 per hr
- Photoperiod: 12 hrs dark / 12 hrs light


Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: Distilled water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Dilution of the test material in distilled water

Duration of treatment / exposure:
Not applicable
Frequency of treatment:
Once
Post exposure period:
24, 48 and 72 h
Doses / concentrations
Remarks:
Doses / Concentrations:
14.7 mL/kg equivalent to 15000 mg/kg
Basis:
nominal conc.
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide
- Route of administration: Oral
- Doses / concentrations: 30 mg/10 mL

Examinations

Tissues and cell types examined:
Bone marrow erythrocytes
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: Preliminary investigations - A few mice were treated orally by various concentrations of the test material diluted with distilled water. Thereby the maximum tolerated dose was estimated at 15 g/kg. At this dosage bone marrow smears showed a reduced number of polychromatic erythrocytes (PCE) as compared with normochromatic erythrocytes (NCE). At a dose exceeding 15 g/kg the mortality was too high.


DETAILS OF SLIDE PREPARATION: Immediately after sacrifice, femurs of a mouse were dissected free of muscle, and by a 1 mL syringe with needle the bone marrow was flushed out into 5 mL of fetal calf serum. After thorough shaking, the mixture was centrifuged for 10 min. at about 1000 rpm. Thereafter, smears were made after removal of the supernatant. The specimens were fixed in methanol and stained with May-Grunwald/Giemsa.


METHOD OF ANALYSIS: Prior to microscopic assessment, all slides were furnished with code numbers, so that the counting was blind. The following counts were made:
Number of normochromatic erythrocytes (NCE) per 1000 erythrocytes
Number of polychromatic erythrocytes (PCE) per 1000 erythrocytes
Number of micronuclei (MN) in 1000 normochromatic erythrocytes
Number of micronuclei (MN) in 1000 polychromatic erythrocytes.

Evaluation criteria:
Increase in the frequency of micronucleated polychromated erythrocytes in treated animals as compared to controls.
Statistics:
The statistical difference was analysed by one-way ANOVA. In the case of PCE (%) the test was performed on the values observed, and for the MN (per thousand) the test was done on computed rank values transformed to normal scores according to Blom's method. (G. Blom Statistical Estimates and Transformed Beta Variables. New York: John Wiley and Sons, Inc., 1958).

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
not specified
Vehicle controls validity:
valid
Negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: Up to 15 g/kg
- Clinical signs of toxicity in test animals: Maximum tolerated dose was estimated at 15 g/kg. At a dose exceeding 15 g/kg, the mortality was too high.
- Evidence of cytotoxicity in tissue analyzed: At 15 g/kg, bone marrow smears showed a reduced number of polychromatic erythrocytes (PCE) as compared with normochromatic erythrocytes (NCE).


RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): No significant difference as compared to controls.
- Appropriateness of dose levels and route: Yes
- Statistical evaluation: Yes

Any other information on results incl. tables

Results:

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
The test material did not induce an increase in the frequency of micronucleated normochromatic erythrocytes in peripheral blood samples from both male and female mice.
Executive summary:

A study was conducted to determine the chromosome-damaging effect of amides, C8-18 and C18-unsatd., N,N-bis(hydroxyethyl) in NMRI mice, according to the method recommended in the First Addendum to OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test) and Method No. 431, Annex V of EEC Directive 79/831.

The experimental animals were 70 NMRI mice, divided into 5 groups. Of the 5 groups three were test groups, one negative control group and one positive control group. The test groups were treated with 15 g test material/kg body weight, the negative control group with distilled water and the positive control group with 30 mg cyclophosphamide/kg body weight. The mice were killed 24, 48 and 72 h respectively after treatment. From bone marrow smears micronucleus counts were made per 1000 polychromatic erythrocytes. The test material did not induce an increase in the frequency of micronucleated normochromatic erythrocytes in peripheral blood samples from both male and female mice.

The test material did induce was non-mutagenic under the conditions of this test.