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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02.03.2010 - 17.05.2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
GLP compliance:
yes (incl. QA statement)
Type of assay:
in vitro mammalian cell gene mutation test using the Hprt and xprt genes

Test material

Constituent 1
Chemical structure
Reference substance name:
3-methylbut-3-en-1-ol
EC Number:
212-110-8
EC Name:
3-methylbut-3-en-1-ol
Cas Number:
763-32-6
Molecular formula:
C5H10O
IUPAC Name:
3-methylbut-3-en-1-ol
Details on test material:
- Name of test material (as cited in study report): 3-Methylbuten-3-ol-1
- Analytical purity: 99%
- Lot/batch No.: 31-0342

Method

Target gene:
HPRT
Species / strain
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Metabolic activation system:
Mammalian liver microsome preparations (S9 mix)
Test concentrations with justification for top dose:
concentrations in μg/mL
Experiment I
without S9 mix: 55.0 110 220 440 880
with S9 mix: 55.0 110 220 440 880

Experiment II
without S9 mix: 55.0 110 220 330 440
with S9 mix: 55.0 110 220 440 880
Vehicle / solvent:
Culture medium
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
7,12-dimethylbenzanthracene
ethylmethanesulphonate
Remarks:
As solvent/vehicle is culture medium, untreated negative controls and negative solvent/vehicle controls are the same.
Evaluation criteria:
A test item is classified as positive if it induces either a concentration-related increase of the mutant frequency or a reproducible and positive response at one of the test points.
Statistics:
A linear regression (least squares) was performed to assess a possible dose dependent
increase of mutant frequencies using SYSTAT􀂣11 (SYSTAT Software, Inc., 501, Canal
Boulevard, Suite C, Richmond, CA 94804, USA) statistics software. The number of mutant colonies obtained for the groups treated with the test item were compared to the solvent control groups. A trend is judged as significant whenever the p-value (probability value) is below 0.05. However, both, biological and statistical significance were considered together.

Results and discussion

Test results
Key result
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.
Therefore, Isoprenol is considered to be non-mutagenic in this HPRT assay.
Executive summary:

The study was performed to investigate the potential of Isoprenol to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster.

The assay was performed in two independent experiments, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a

treatment period of 4 hours. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.

The highest concentration of the test item (880 μg/mL) was equal to a molar concentration of about 10 mM.

The tested concentrations are described in Table 2 (page 19). The evaluated experimental points and the results are summarised in Table 1 (page 13).

No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments.

Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test item and the activity of the metabolic activation system.