3-methoxy-3-methylbutan-1-ol

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:

2012-08-17 to 2012-09-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

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:

3-methoxy-3-methylbutan-1-ol

EC Number:

260-252-4

EC Name:

3-methoxy-3-methylbutan-1-ol

Cas Number:

56539-66-3

Molecular formula:

C6H14O2

IUPAC Name:

3-methoxy-3-methylbutan-1-ol

Test material form:

other: liquid

Details on test material:

- Name of test material: 3-methoxy- 3-methylbutan-1-ol (MMB)
- Physical state: liquid
- Analytical purity: 99.47% (GC area %)
- Lot No.: 32726
- Expiration date of the lot: June 30, 2013

Method

Target gene:

Thymidine Kinase (TK+/-)

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media: complete growth medium F10P [Dulbecco’s modified Eagle’s Medium (DMEM) with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplied with 0.1 % Pluronic, 90 % and Fetal bovine serum, 10 %]. Treatment medium F5P [Dulbecco’s modified Eagle’s Medium (DMEM) with 4 mM Lglutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplied with 0.1 % Pluronic, 95 % and Fetal bovine serum, 5 %].

Metabolic activation:

with and without

Metabolic activation system:

A co-factor supplemented postmitochondrial fraction (S9) from the liver of Aroclor 1254 induced Sprague-Dawley rats

Test concentrations with justification for top dose:

0.50, 0.25, 0.125 and 0.0625 mg/ml

Controls

Untreated negative controls:

yes

Remarks:

(sterile distilled water)

Negative solvent / vehicle controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

methylmethanesulfonate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 13 days

SELECTION AGENT (mutation assays): Trifluorothymidine (TFT)

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED: 3 x 10E6

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

The total number of colonies per TFT plate were determined for those cultures with >= 10% total growth.

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 nor precipitates, but tested up to recommended limit concentrations

Remarks:

Cytotoxicity was observed at the highest concentration tested (0.5 mg/ml).

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: The testing concentrations of the test substances for the study were selected based on the results of preliminary range-finding test using L5178Y/TK+/- mouse lymphoma cells (non- GLP study 7191037866-02 conducted from 18 to 26 July 2012). In the range-finding test, cytotoxic effect of cell growth inhibition was observed at 0.5 mg / ml and above concentrations. Based on the results of range-finding test, four concentrations were selected, which were 0.50 mg/ml, 0.25 mg/ml, 0.125 mg/ml and 0.0625 mg/ml, respectively.

Applicant's summary and conclusion

Conclusions:

From the above study, the test item 3-methoxy-3-methylbutan-1-ol (MMB) is non-mutagenic to the L5178Y TK+/- clone, both in the absence and presence of S9 metabolic activation system.