Methyl 3-methoxypropionate

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:

1989-1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to international guideline. All the required bacterial strains were used in this study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Methyl 3-methoxypropionate

EC Number:

223-358-1

EC Name:

Methyl 3-methoxypropionate

Cas Number:

3852-09-3

Molecular formula:

C5H10O3

IUPAC Name:

methyl 3-methoxypropanoate

Details on test material:

- batch number: 89227-M3
- appearance: colourless liquid
- storage conditions: stored at room temperature
- expiry date: stable for at least one year from date of manufacture on 27/2/89

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

312.5, 625, 1250, 2500, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

other: 2-aminoanthracene

Details on test system and experimental conditions:

- without metabolic activation:
0.1 ml aliquots of bacterial suspension and 0.5 ml of sterile 0.1 M sodium orthophosphate buffer (pH 7.4) were added to each of one set of sterile bijou bottles

0.1 ml of the test compound was added to cultures at 5 concentrations seperated by a two-fold interval. 3 bottles were used at each dose level.

2.0 ml of histidine/tryptophan deficient agar were added to each of the bottles, thoroughly mixed and then overlaid onto previously prepared plates containing 25 ml of minimal agar. Plates were incubated for 3 days at 37˚C.

Colonies were counted using a Biotran Automatic Colony Counter, and the mean number of revertant colonies per treatment group assessed.

- with metabolic activation:
Methodology was as described above except that 0.5 ml of liver homogenate S-9 mix was added to bijou bottles in place of sterile buffer.

Evaluation criteria:

1) If treatment with a test material produces an increase in revertant colony numbers of at least twice the concurrent solvent controls, with some evidence of a positive dose-relationship, in 2 separate experiments, with any bacterial strain eigher in the presence or absence of S9 mix, it is considered to show evidence of mutagenic activity in this test system. No statistical analysis is performed.
2) If treatment with a test material does not produce reproducible increases of at least 1.5x the concurrent solvent controls, at any dose level with any bacterial strain, it is considered to show no evidence of mutagenic activity in this test system. No statistical analysis is performed.
3) If the results fail to satisfy the criteria for a clear positive or negative response, the following approach is taken:
- Repeat tests may be performed using modifications of the experimental method. These modifications include but are not restricted to, the use of a narrower dose range than that already tested; the use of different levels of liver homogenate S9 fraction in the S9 mix. Should an increase in revertant colony numbers be observed which satisfies paragraph 1) the material is considered to show evidence of mutagenic activity in this test system. No statistical analysis is performed.
- If no clear "positive" response can be obtained the test data may be subjected to analysis to determine the statistical significance of any observed increases in revertant colony numbers. The statistical procedure is normally analysis of variance followed by Student's t test.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

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

When tested at dose levels up to 5000 µg/plate, methyl-3-methoxypropionate was not mutagenic in this study.

Executive summary:

Methyl-3 -methoxypropionate was not toxic towards the tester strains in the preliminary toxicity test. 5000 µg/plate was chosen as the top dose level in the mutation tests.

In the mutation tests, no substantial increases in revertant colony numbers of any of the tester strains were observed following treatment level with methyl-3 -methoxypropionate at any dose level, either in the presence or absence of S9 mix.