Reaction mass of ethyl (2S,3S)-3,4-dibromo-2-methylbutanoate and ethyl (2R,3R)-3,4-dibromo-2-methylbutanoate and ethyl (2S,3R)-3,4-dibromo-2-methylbutanoate and ethyl (2R,3S)-3,4-dibromo-2-methylbutanoate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-08-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well-documented study report which meets basic scientific methods; not a guideline method. Method based on modification of OECD TG 471.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

Ames II

Deviations:

yes

Remarks:

Ames II

Principles of method if other than guideline:

The test method is used to evaluate the mutagenic potential of the test substance a based on the ability to induce point mutations in selected loci of several strains of Salmonella typhimurium in a modified version of the Ames test (OECD 471), designated Ames II Assay (microtiter version), both with and without the addition of a metabolizing system (S9 mix) obtained from induced rat liver. According to the authors, this method shows a good accurancy concerning the prediction of the results in the regular Ames test.

GLP compliance:

yes

Remarks:

Sceening in GLP-certified Laboratory

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

ethyl (2R,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2R,3S)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3S)-3,4-dibromo-2-methylbutanoate

EC Number:

700-225-0

Cas Number:

1160806-44-9

Molecular formula:

C7 H12 Br2 O2

IUPAC Name:

ethyl (2R,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2R,3S)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3R)-3,4-dibromo-2-methylbutanoate; ethyl (2S,3S)-3,4-dibromo-2-methylbutanoate

Details on test material:

- Name of test material (as cited in study report): 2-Methyl-3,4-dibrombuttersäureethylester
- Physical state: liquid, colorless, clear
- Analytical purity: 98.1 wt%
- Lot/batch No.: 35684-12-5; date of production: 2009-07-14
- Storage condition of test material: refrigerator, under light exclusion
No further data.

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 98

Species / strain 2

Species / strain / cell type:

S. typhimurium, other: TA mix (TA 7001 - TA 7006)

Metabolic activation:

with and without

Metabolic activation system:

induced rat liver S9

Test concentrations with justification for top dose:

0, 4, 20, 100, 500, 2500, 5000 µg/ml

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: no data

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: without S9: 4-nitrofluorene (5 µg/ml) + 2-nitrofluorene (100 µg/ml); with S9: 2-aminoanthracene (125 µg/ml)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in suspension; liquid fluctuation test with and without induced rat liver S9 mix

NUMBER OF REPLICATIONS: 48 wells in triplicate plates per dose or per control

DETERMINATION OF CYTOTOXICITY
- Method: clearing of the background lawn, decrease in the number of yellow wells

Evaluation criteria:

no data

Statistics:

no data

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium, other: TA mix (TA 7001 - TA 7006)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

MUTAGENICITY:
A relevant increase in the number of positive wells (his+ revertants) was not observed either without S9 mix or after the addition of a metabolizing system.

TEST-SPECIFIC CONFOUNDING FACTORS:
- Precipitation: No precipitation was observed.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
No bacteriotoxic effect (clearing of the background lawn, decrease in the number of yellow wells) was observed up to the highest applied concentration.

For details, see attached files.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the experimental conditions chosen here, it is concluded that 2-Methyl-3,4-dibrombuttersäureethylester is not a mutagenic substance in the Ames II Assay (Salmonella typhimurium reverse mutation assay) in the absence and the presence of metabolic activation.