3-(Hydroxymethyl)-1-pentene

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 04, 2006 - April 17, 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

07-1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)

Version / remarks:

06-2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian cell gene mutation tests using the thymidine kinase gene

Test material

Test material information

Constituent 1

Reference substance name:

2-ethylbut-3-en-1-ol

EC Number:

610-949-8

Cas Number:

53045-70-8

Molecular formula:

C6H12O

IUPAC Name:

2-ethylbut-3-en-1-ol

Test material form:

liquid

Method

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9 mix of Aroclor 1254 induced animals

Test concentrations with justification for top dose:

Experiment 1: 0, 15.8, 50, 158, 500, 1580 and 5000 µg/ml (+/- S9 mix)
Experiment 2: 0, 158, 500, 1580 and 5000 (+/- S9 mix)

Vehicle / solvent:

Ethanol

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

7,12-dimethylbenzanthracene

Remarks:

Positive control for experiments without S9 mix: NQO Positive control for experiments with S9 mix: DMBA

Details on test system and experimental conditions:

In a preceding range finding test, the relative survival was determined after exposure to various test material concentrations ranging between 5 and 5000 μg/mL. A clear reduction in the relative survival of the cells occurred at the concentration of 5000 μg/mL in the absence and presence of S9 mix, respectively. Precipitation of the test item in the cell culture medium was not seen. A relevant change in the pH and the osmolarity of the culture medium was not detected in the concentration range tested in the main study.

Evaluation criteria:

see below

Statistics:

All calculations such as determination of survival or viability, determination of mutant frequency or assessment of statistical significance of mutant frequency were performed by computer using validated software.

Results and discussion

Test results

Key result

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5000 ug/mL in the 1st series without S9

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

In this GLP study performed according to OECD GL 476, the test item was assessed for its ability to induce mutation at the tk locus (5-trifluorothymidine resistance) in mouse lymphoma cells using a fluctuation protocol. Based on the obtained results and under the conditions employed in this study, the test item was not mutagenic in this test system both in the absence or presence of S9 mix.

Executive summary:

Study design

In this GLP study performed according to OECD GL 476 the test item was assayed for its ability to induce mutations at the TK locus (5-trifluorothymidine resistance) in L5178Y mouse lymphoma cells using a fluctuation protocol. The study consisted of two independent experimental series, each conducted in the absence and presence of an exogenous metabolizing system (S9 mix from livers of rats pre-treated with Aroclor 1254). Ethanol was used as the solvent. The exposure times were 3 and 24 hours in the absence and 3 hours in the presence of S9 mix, respectively. Test item concentrations ranging from 15.8 to 5000 μg/mL were tested in the absence or presence of S9 mix.

 

Results

No precipitation of the test material in the incubation medium was observed. Clear cytotoxic effects, i.e. a relevant decrease in either the % relative survival or % total growth of the test cells, occurred at the highest concentration tested in the 1st experiment in the absence of S9, i.e. at 5000 μg/mL. The doses tested were selected to determine viability and mutagenicity (5 -trifluorothymidine (TFT) resistance) 2 days after treatment. Negative (solvent) and positive control treatments were included in each mutation experiment in the absence and presence of S9 mix. Mutant frequencies in negative control cultures fell within normal ranges, and clear increases in mutation were induced by the positive control chemicals 4-nitroquinoline N-oxide (without S9 mix) and 7, 12-dimethylbenz[a]anthracene (with S9 mix). Therefore, the study was accepted as valid.

 

Conclusion

No relevant increases in mutant frequency were observed following treatment with test item in the two experimental series in the absence and presence of S9 mix. It is therefore concluded that the test item is non-mutagenic in this test system under conditions where the positive controls exerted potent mutagenic effects.