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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:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Study period:
27 Nov 1984 -20 Dec 1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study with acceptable restrictions. Lack of test material details. Only the highest dose group was evaluated.

Data source

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

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
yes
Remarks:
lack of test material details; only the highest dose group was evaluated
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): only trade name given
- Chemical denomination: Glyceride, C16-18 und C18 mono- und dihydroxyfettsäuren
- Physical state: white solid
- Analytical purity: no data
- Analytical data: melting point: 94 °C; acid value: 1.0; saponification value: 178; hydroxyl value: 138; iodine value: 3.4; epoxy oxygen content: 0.4%

Test animals

Species:
mouse
Strain:
other: CFW 1 (Winkelmann)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: 8 weeks
- Weight at study initiation: 21-29 g
- Fasting period before study: animals were fasted at least 4 h prior to administration.
- Housing: male animals were housed individually in Makrolon cages type I, female animals were housed in groups up to 4 animals in Makrolon cages type II
- Diet: Haltungsdiät Altromin 1324, 10 mm pellets (Altrogge Spezialfutter), feeding frequency not further specified
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: peanut oil Oleum Arachidis G166/DAB 8
- Amount of vehicle: 20 mL/kg bw
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
An aliquot of the test substance was molten at 70 ºC and mixed with 70 ºC warm peanut oil. After cooling of the solution, the mixture was applicated by oral gavage.

Duration of treatment / exposure:
single application
Frequency of treatment:
single application
Post exposure period:
24, 48 and 72 h after treatment
Doses / concentrations
Remarks:
Doses / Concentrations:
1000, 5000 and 10000 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
Preliminary study: 2
Main study: 7
Control animals:
yes, concurrent vehicle
Positive control(s):
- Cyclophosphamide
- Route of administration: intraperitoneal injection
- Doses / concentrations: 10 mg/kg bw in water

Examinations

Tissues and cell types examined:
Tissue: femoral bone marrow
Cell type: erythrocytes of the bone marrow
Details of tissue and slide preparation:
DETAILS OF SLIDE PREPARATION:
Slides were fixed with methanol and stained with Giemsa-solution for 10 min.

METHOD OF ANALYSIS: 1 slide per animal was analysed. The number of micronuclei was counted in 1000 polychromatic erythrocytes per animal and the ratio of polychromatic and normochromatic erythrocytes was recorded.

Statistics:
Mean values and standard deviations were calculated. For calculation of significances the tables of Kastenbaum and Bowman (Kastenbaum, M.A. and Bowman, K.O., 1970, Mutation Research 9:527-549) were used.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Remarks:
Animals of all dose groups showed reduced activity and ruffled fur.
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 5000, 7500 and 10000 mg/kg bw
- Clinical signs of toxicity in test animals: In the mid dose-group slightly reduced activity and ruffled fur was observed up to 20 h after application. In the high dose-group all animals showed slightly reduced activity, in male animals strongly ruffled fur and in female animals slightly ruffled fur was observed. No clinical signs of systemic toxicity were observed after 48 h.

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei: no significant increase of micronuclei number was observed
- Ratio of PCE/NCE: no influence of the test material was observed in comparison to the controls
- Clinical signs of toxicity in test animals: In the low dose-group slightly reduced activity after application was observed and in the mid dose-group slightly reduced activity and ruffled fur was observed after application. No clinical signs of toxicity were observed after 24 h. In the high dose-group all animals showed slightly reduced activity and strongly ruffled fur. No clinical signs of systemic toxicity were observed after 20 h.

Any other information on results incl. tables

No deaths occured during the study. As there were no indications for genotoxicity in animals dosed with 10000 mg/kg bw, analysis of micronuclei was not performed in animals dosed with 1000 and 5000 mg/kg bw.

Table 1: Results of the in vivo micronucleus assay in male animals

Exp group

Number of animals

Sampling time (h after administration)

Dose

[mg/kg bw]

Mean

PCEs/NCEs at sampling time

Total micronuclei per 1000 PCEs at sampling time

mean values

range

Vehicle control

(peanut oil)

7

24

20 mL/kg

1.11

1.57

0-2

Positive control

(cyclophosphamide)

7

24

10

1.10

7.71

4-11

Test substance

7

24

10000

1.20

2.14

1-4

Test substance

7

48

10000

1.00

1.00

0-2

Test substance

7

72

10000

1.25

1.43

0-3

  

Table 2: Results of the in vivo micronucleus assay in female animals

Exp group

Number of animals

Sampling time (h after administration)

Dose

[mg/kg bw]

Mean

PCEs/NCEs at sampling time

Total micronuclei per 1000 PCEs at sampling time

mean values

range

Vehicle control

(peanut oil)

7

24

20 mL/kg

1.26

1.14

0-3

Positive control

(Cyclophosphamide)

7

24

10

1.15

6.86

4-10

Test substance

7

24

10000

1.07

2.00

1-4

Test substance

7

48

10000

1.06

2.29

0-6

Test substance

7

72

10000

1.15

1.86

0-3

PCE = polychromatic erythrocytes

NCE = norchromatic erythrocytes

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative