Amides, C16-18 and C18-unsatd., N,N-bis(hydroxyethyl)

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Read across study hence maximum reliability rating of 2 assigned according to ECHA guidance, although study was conducted according to OECD Guideline 474

Justification for data waiving:

other:

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

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

Test material information

Constituent 1

Reference substance name:

Amides, C8-18 and C18-unsatd., N,N-bis(hydroxyethyl)

EC Number:

268-935-9

EC Name:

Amides, C8-18 and C18-unsatd., N,N-bis(hydroxyethyl)

Cas Number:

68155-07-7

IUPAC Name:

68155-07-7

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 or test system 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/ h
- Photoperiod: 12 h dark / 12 h 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 15 g/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

None

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
Under the test conditions, structurally similar test substance 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 structurally similar 'amides, C8-18 (even-numbered) 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.

 

Under the test conditions, structurally similar test substance did not induce an increase in the frequency of micronucleated normochromatic erythrocytes in peripheral blood samples from both male and female mice.