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Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

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Administrative data

in vitro gene mutation study in bacteria
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May-June 1995
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: study performed in accordance with the corresponding OECD-/EU-testing guidelines
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Methyl acetoacetate
EC Number:
EC Name:
Methyl acetoacetate
Cas Number:
methyl 3-oxobutanoate
Details on test material:
- Physical state: Clear colourless liquid
- Storage condition of test material: Room temperature, protected from light


Target gene:
not applicable
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9-liver fractions of Sprague-Dawley rats, induced with Aroclor 1254
Test concentrations with justification for top dose:
Test substance concentrations of 50, 150, 500, 1500 and 5000 microg/plate were used in the main test.
Vehicle / solvent:
- Vehicle: Acetone
- Justification for choice of solvent/vehicle: high solubility, better than others
Untreated negative controls:
each strain alone or in presence of the solvent
Negative solvent / vehicle controls:
True negative controls:
Positive controls:
Positive control substance:
further positive control substances were used: 9-Aminoacridine, 2-Nitroquinoline-oxide, 2-Aminoanthracene
Details on test system and experimental conditions:
The study was performed by the direct plate incorporation method with and without metabolic activation.
Evaluation criteria:
For a substance to be considered positive in this test system, it should have induced a dose-related and statistically significant increase in mutation rate (of at least twice the spontaneous reversion rate) in one or more strains of bacteria in the presence and/or absence of the S9 microsomal enzymes in both experiments at sub-toxic dose levels. If the two experiments give conflicting results or equivocal results are obtained, then a third experiment may be used to confirm the correct response.
To be considered negative the number of induced revertants compared to spontaneous revertants should be less than twofold at each dose level employed, the intervals of which should be between 2 and 5 fold and extend to the limits imposed by toxicity, solubility or up to the maximum recommended dose of 5000 ug/plate. In this case the limiting factor was the maximum recommended dose.
According to the OECD guideline 471, a statistical analysis of the data is not mandatory.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
Untreated negative controls validity:
Positive controls validity:
Additional information on results:
No precipitation observed.
Remarks on result:
other: all strains/cell types tested
Migrated from field 'Test system'.

Applicant's summary and conclusion

Interpretation of results (migrated information):
negative with and without metabolic activation

It can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations in the five strains tested.
Executive summary:

The assay was performed 1995 in compliance with GLP. The test was performed with and without liver microsomal activation and used strains TA 1535, TA 1537, TA 98, TA 100 and E.coli WP2uvrA. Each concentration and the controls, were tested in triplicate.

The test item was tested at the following concentrations: 50; 150; 500; 1500 and 5000 µg/plate (pre-experiment and main test). The plates incubated with the test item showed normal back­ground growth up to 5000 µg/plate with and without S9 mix in all strains used. No toxic effects, evident as a reduction in the number of revertants occurred in the test groups with and without metabolic activation. Precipitation was not observed.

Valeronitrile was found to be non-mutagenic in this test system.