Ferrocene

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 1987 (no further details given)

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:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

not specified

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Ferrocene

EC Number:

203-039-3

EC Name:

Ferrocene

Cas Number:

102-54-5

Molecular formula:

C10H10Fe

IUPAC Name:

iron(2+) dicyclopenta-2,4-dienide

Details on test material:

- Name of test material (as cited in study report): Ferrocene
- Substance type: technical product
- Physical state: red-brown solid
- Analytical purity: no data
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: C10-H10-Fe
- Isomers composition: no data
- Purity test date: no data
- Lot/batch No.: no data
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: no data

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann Co. Versuchstierzucht GmbH & Co. KG, 4799 Borchem/Kreis Paderborn
- Age at study initiation: about 10 weeks
- Weight at study initiation: 28 g (males); 25 g (females)
- Assigned to test groups randomly: no data
- Fasting period before study: no data
- Housing: no data
- Diet (e.g. ad libitum): 3 g/day of maintenance feed for rats and mice
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25
- Humidity (%): no data
- Air changes (per hr): "continuous air circulation"
- Photoperiod (hrs dark / hrs light): no data

IN-LIFE DATES: From: no data

Administration / exposure

Route of administration:

other: gavage or in feed

Vehicle:

- Vehicle(s)/solvent(s) used: maize germ oil
- Justification for choice of solvent/vehicle: commonly used vehicle in animal studies
- Concentration of test material in vehicle: 70 mg/5 ml vehicle
- Amount of vehicle (if gavage or dermal): 0.2 ml
- Lot/batch no. (if required): no data
- Purity: no data

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: calculated from body weight to give a dose of 100 mg/kg bw

Duration of treatment / exposure:

Single exposure then sampled at 24, 48 and 72 hours

Frequency of treatment:

once

Post exposure period:

24, 48 and 72 h

Doses / concentrations

Remarks:

Doses / Concentrations:
100 mg/kg bw
Basis:
actual ingested
(given by gavage)

No. of animals per sex per dose:

5/sex

Control animals:

yes, concurrent no treatment

Positive control(s):

Endoxane (cyclophosphamide)
- Justification for choice of positive control(s): frequently used in the test laboratory
- Route of administration: gavage
- Doses / concentrations: 100 and 250 mg/kg bw

Examinations

Tissues and cell types examined:

Bone marrow erythrocytes

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: maximum tolerated dose

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): treated by gavage with 100 mg/kg bw test substance; bone marrow cells sampled 24, 48 and 72 h later. Positive control sampled at 24 h only.

DETAILS OF SLIDE PREPARATION: bone marrow suspension in 2 ml fetal calf serum centrifuged, 1.8 ml of the serum was discarded and the pellet resuspended in the remaining serum. 5 ul was applied to slides and smears pepared using an automatic smear device. The smers were air-dried for 24 h before staining with May-Grunwald and Giemsa.

METHOD OF ANALYSIS: smears were microscopically examined and the number of micronuclei per 1000 polychromatic erythrocytes were recorded. The ratio of monochromatic to polychromatic erythrocytes was determined for 1000 erythrocytes.

Evaluation criteria:

no data

Statistics:

no data

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative

In a reliable study, ferrocene exhibited no potential to induce micronuclei in the bone marrow of mice (5/sex) at the maximum tolerated dose of 100 mg/kg bw (given as a single dose by gavage) in an in vivo mammalian micronucleus assay.

Executive summary:

In a reliable study, ferrocene was assessed for its ability to induce chromosome damage in an in vivo bone marrow micronucleus assay in mice, carried out in accordance with EU test method B.12.

Groups of five mice of each sex were gavaged with 100 mg/kg bw of the test substance and the bone marrow was sampled 24, 48 and 72 h later. After concentrating the cells by centrifugation, smears were prepared on glass slides, stained and scored for micronuclei. The number of micronuclei per 1000 polychromatic erythrocytes was recorded and the ratio of monochromatic to polychromatic erythrocytes was calculated.

There was no increase in micronuclei frequency in either male or female mice when compared to the control animals. The positive control gave the expected result, confirming the sensitivity of the assay.

In a reliable study, ferrocene exhibited no potential to induce micronuclei at the maximum tolerated dose of 100 mg/kg bw in an in vivo bone marrow micronucleus assay in mice.