1,1'-isopropylidenebis(ethylferrocene)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation 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

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Description : Orange coloured crystalline solid
Date received : 03 April 2012
Expiry date : 03 April 2013
Storage conditions : Room temperature, in the dark, under nitrogen

Method

Target gene:

thymidine kinase, tk +/- locus of the L5178Y mouse lymphoma cell line

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media: Cells were routinely cultured in RPMI 1640 medium with Glutamax-1 and HEPES buffer (20 mM) supplemented with Penicillin (100 units/ml), Streptomycin (100 μg/ml), Sodium pyruvate (1 mM), Amphotericin B (2.5 μg/ml) and 10% donor horse serum (giving R10 media) at 37 °C with 5% CO2 in air
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes

Metabolic activation:

with and without

Metabolic activation system:

S9-mix was prepared immediately prior to dosing by mixing S9, NADP (5 mM), G-6-P (5 mM), KCl (33 mM) and MgCl2 (8 mM) in R0

Test concentrations with justification for top dose:

For Experiment 1 the dose range was 29.06 to 930 µg/ml in the absence of S9 and 1.82 to 116.25 µg/ml in the presence of S9. In Experiment 2 the dose range was 0.25 to 12 µg/ml in the absence of S9 and 2 to 80 µg/ml in the presence of S9

Vehicle / solvent:

dimethyl sulfoxide

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

Experiment 1&2 absence of metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

Experiment 1&2 presence of metabolic activation

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: strain/cell type: thymidine kinase, tk +/-

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

It is concluded that dicyclopentadienyl iron did not induce mutation at the tk locus of L5178Y mouse lymphoma cells when tested under the conditions employed in this study. These conditions included four independent treatments at concentration up to 32 μg/ml in the presence (4 hours) of a rat liver metabolic activation system (at 1 and 2% (v/v) final concentration of S9 fraction) and at concentrations of 232.5 µg/ml and 2 μg/mL in the absence (4 and 24 hours, respectively) of metabolic S9. The maximum doses tested were limited by either acceptable reductions in toxicity (as measured by %RTG) or by precipitate (observed by eye) at the end of treatment in the absence or presence of S9, respectively.

Executive summary:

Ferrocene was assessed for its ability to induce gene mutations in an in vivo assay using mouse lymphoma L5178Y cells and conducted according to OECD Guideline 476.

It is concluded that dicyclopentadienyl iron did not induce mutation at the tk locus of L5178Y mouse lymphoma cells when tested under the conditions employed in this study. These conditions included four independent treatments at concentration up to 32 μg/ml in the presence (4 hours) of a rat liver metabolic activation system (at 1 and 2% (v/v) final concentration of S9 fraction) and at concentrations of 232.5 µg/ml and 2 μg/mL in the absence (4 and 24 hours, respectively) of metabolic S9. The maximum doses tested were limited by either acceptable reductions in toxicity (as measured by %RTG) or by precipitate (observed by eye) at the end of treatment in the absence or presence of S9, respectively.