Fusel oil

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / micronucleus study

Remarks:

Type of genotoxicity: DNA damage and/or repair

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable publication which meets basic scientific principles

Data source

Reference

Reference Type:

publication

Title:

Evaluation of the genotoxic potential of some microbial volatile organic compounds (MVOC) with the comet assay, the micronucleus assay and the HPRT gene mutation assay

Author:

Kreja L and Seidel HJ

Year:

2002

Bibliographic source:

Mutation Research 513: 143-150

Materials and methods

Principles of method if other than guideline:

The study was conducted according to the method described by Miller BM et al. (1995). Environ. Mol. Mutagen 26: 240-247.

GLP compliance:

not specified

Type of assay:

in vitro mammalian cell micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

3-methylbutan-1-ol

Cas Number:

123-51-3

Molecular formula:

C5H12O

IUPAC Name:

3-methylbutan-1-ol

Details on test material:

- Name of test material (as cited in study report): 3-methyl-1-butanol
- Source: Fluka
- Analytical purity: no data

Method

Target gene:

not applicable

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media: in 75 cm2 tissue culture flasks at 37°C in a humidified atmosphere with 5% C02 in MEM Eagle medium supplemented with 10% heat-inactivated fetal calf serum (FCS), 2 mM L-glutamine, 100 IU/ml penicillin and streptomycin.

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Arcolor 1254-induced rat liver S9-mix

Test concentrations with justification for top dose:

5, 9 and 23 mM

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO (1% maximal concentration)
- Justification for choice of solvent/vehicle: was shown to produce no genetic toxicity effect

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

without metabolic activation Migrated to IUCLID6: 0.25 and 0.5 mM

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

with metabolic activation Migrated to IUCLID6: 0.1 mM

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: Cells were cultivated on microscope slides in quadriperm-dishes. A total of 1E+05 cells were seeded in each chamber and cultivated 24 hours before treatment. The medium was then substituted by 6 ml of fresh medium containing the test compound, and the cells were incubated for 4 hours. For experiments with metabolic activation, the test medium additionally contained 3% of S9-mix.
- Expression time (cells in growth medium): After treatment the cultures were washed twice with medium and incubated further for 24 hours.

NUMBER OF CELLS EVALUATED: Micronuclei (MN) were analyzed in 1000 cells/culture. To show reproducibility of the results 2 to 4 independent experiments were performed, and the mean MN frequency per 100 cells ± SD, was calculated.

DETERMINATION OF CYTOTOXICITY
no data

Evaluation criteria:

The test compound was classified as a clastogen when it was able to enhance the spontaneous MN frequency at least 3-fold or higher over that of the control for at least one dose tested.

Statistics:

Differences between the non-exposed control and the test substance exposed to V79 cells were tested for significance (P < 0.05) using the Student's t-test.

Results and discussion

Test results

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

A concentration-dependent significant increase of micronuclei was found after MMS exposure (t-test; P< 0.001). A level of 0.5% spontaneous MN was found in the non-exposed control sample. After cyclophosphamide treatment used as a positive control for S9-mix metabolic activation, MN frequency was increased from 0.3 to 4.8% (single experiment). However, after exposure to the test substance the MN frequency was not significantly increased.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Mean MN frequencies ± SD calculated from 2 to 4 experiments are shown in the table:

 

Test substance	Concentration (mM)	Micronucleus frequency (%)^a
		Without S-9-mix	With S-9-mix
Control non-exposed		0.5 ± 0.3
Cyclophosphamide	0.1	0.3	4.8
MMS	0.25	3.4 ± 1.1
	0.5	11.8 ± 4.4
3-Methyl-1-butanol	5	0.3 ± 0.1	1.0 ± 0.1
	9	0.8 ± 0.3	0.4
	23	0.7 ± 0.4
DMSO	350	0.21 ± 0.2	1.6 ± 1.9
	700	0.9 ± 0.2	1.1 ± 1.0
^aMean values from 2 to 4 experiments ± SD

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative