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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
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1989/10/17- 1990/01/30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
(draft)
Deviations:
no
GLP compliance:
yes
Type of assay:
other: micronucleus assay

Test material

Constituent 1
Reference substance name:
151661-88-0
Cas Number:
151661-88-0
IUPAC Name:
151661-88-0
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Specific details on test material used for the study:
- Name of test material: Stabiol VP 1711
- Substance type: Fatty acid C18 and C18-unsaturated epoxidized, ester with ethylene glycol
- Physical state: white powder
- Lot/batch No.: Nr. 41, 041/8/055
- Stability under test conditions: not given
- Storage condition of test material: at room temperature, at 20 °C the test item only hydrolyses 1 % /day

Test animals

Species:
mouse
Strain:
other: CFW 1
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, 4799 Borchen, Germany
- Age at study initiation: Aged 7 to 8 weeks
- Weight at study initiation: Between 20.7 g and 32.8 g for the males; between 19.5 g and 24.8 g for the females.
- Housing: Male mice were housed individually macrolon cages type I; female mice were housed in groups up to three in macrolon cages type II.
- Diet: Altromin No. 1314 , 10 mm pellet diameter, supplied from Altrogge Spezialfutter.
- Water: Drinking water
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21°C
- Humidity: 30 - 70 %
- Photoperiod: 12 hours dark/ light cycle

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle: arachis oil
- Justification for choice of solvent/vehicle: solubility of the test item and not toxic to rats.
- Concentration of test material in vehicle: 3000, 4000, 5000 mg/kg bw for the dose range finding study, 5000 mg/kg bw for the main trial.
- Amount of vehicle: 10 mL/kg
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Solutions were prepared on the day of administration. The stability of CP at room temperature is good. In aqueous solution at 20°C only 1 % of CP is hydrolyzed per day.
Duration of treatment / exposure:
24, 48 or 72 hours
Frequency of treatment:
once
Post exposure period:
none
Doses / concentrations
Dose / conc.:
5 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
dose range finder: 3 groups of 2 males and 2 females
main trial: 5 groups of 6 males and 6 females
Control animals:
yes, concurrent vehicle
Positive control(s):
- Justification for choice of positive control: Cyclophosphamide was selected due to its genotoxic potential after metabolic activation and is recommended as positive control in the guideline.
- Route of administration: Intraperitoneal administration
- Doses/ concentrations: 10 mg/kg bw in bidistilled water

Examinations

Tissues and cell types examined:
bone marrow cells
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: based on a dose range finding study

TREATMENT AND SAMPLING TIMES: The animals of all treatment groups received a single dose at a volume of 10 mL/kg bw

DETAILS OF SLIDE PREPARATION:
Bone marrow smears from both femurs of each animal were prepared. The slides were air-dried at least overnight and then stained with Giemsa according to modification of Gollapudi and Kamra. The slides were fixed in 100 % methanol for 5 min., then rinsed twice in bidistilled water and thereafter stained in Giemsa solution. After air -drying the back side was cleaned, necessary with ethanol and then dipped for 3 min. in xylol.

METHOD OF ANALYSIS:
From the three slides prepared per animal, one slide was chosen and randomly scored. The slides of five male and five female animals per treatment group were scored microscopically at a magnification of 1000. The number of micronucleated cells was counted in 1000 polychromatic erythrocytes/ per animal. The ratio of polychromatic to normochromatic erythrocytes was determined by counting and differentiating the first 1000 erythrocytes at the same time. Averages and standard deviations were calculated after decoding the complete scoring results.
Evaluation criteria:
- A micronucleus test is considered acceptable if it meets the following criteria: the positive control substance induced a statistical significant increase in the frequency of micronucleated polychromatic erythrocytes- the incidence of micronuclei should reasonably fall within the historical control data range of the laboratory.
- A test item is considered positive in the micronucleus test if it induced a biologically as well as statistically significant increase (p < 0.05) in the frequency of micronuclei at any dose or at any sampling either in the male or in the female groups.
- A test substance is considered negative in the micronucleus test if none of the tested doses or sampling times showed a statistically significant (p < 0.05) increase in the incidence of micronuclei, neither in male nor in female groups.
Statistics:
Statistical analysis of data was performed by calculating the statistical significance versus negative controls with the aid of the tables of Kastenbaum and Bowman. The preceding criteria are not absolute and other factors may influence the final evaluation decision.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: the dose range finding study determined 5000 mg/kg bw as final concentration for the main trial.
- Clinical signs of toxicity in test animals: no further signs of toxicity were observed.
- Rationale for exposure: based on the non toxic effect in the dose range finding study.

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei: no
- Ratio of PCE/NCE: mean of animal 1 - 5 at a dose of 5000 mg/kg bw = 1.24
- Statistical evaluation: The investigated test item did not induce a statistically significant (time-dependent) increase in the number of micronucleated polychromatic erythrocytes in the bone marrow of male or female mice.

Applicant's summary and conclusion