Ethyl cyanoacetate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP guideline study. Ethyl cyanoacetic acid and 2-ethylhexyl cyanoacetic acid are both esters of cyanoacetic acid, both hydrolyze rapidly under neutral and alkaline conditions to cyanoacetic acid and ethanol or 2-ethylhexanol, respectively. It ca be assumed that for most endpoints cyanoacetic acid will be the common metabolite that determines the toxicity of both substances.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-ethylhexyl cyanoacetate

EC Number:

236-425-5

EC Name:

2-ethylhexyl cyanoacetate

Cas Number:

13361-34-7

Molecular formula:

C11H19NO2

IUPAC Name:

2-ethylhexyl cyanoacetate

Details on test material:

- Name of test substance: 2-ethylhexyl-cyanoacetate
- Substance number: 96/219
- Batch number: ABL. NR. 13-9844
- CAS number: 13361-34-7
- Degree of purity/content: 98.5 g/100 g (GC)
- Date of manufacturing: 27-DEC-1995
- Physical state/appearance: liquid, colorless to light yellowish
- Stability under test conditions: The stability of the TS over the study period has been proven by reanalysis. The stability of the TS in the vehicle DMSO and in water was assured analytically.
- Storage conditions: room temperature

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:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9 mix

Test concentrations with justification for top dose:

Standard plate test: 20, 100, 500, 2500 and 5000 µg/plate
Preincubation test: 20, 100, 500, 1000 and 2000 µg/plate

Vehicle / solvent:

- Vehicle/solvent used: DMSO

Controls

Untreated negative controls:

yes

Remarks:

sterility control

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

With S-9: 2.5 µg 2-AA (TA 100, TA 98, TA 1537, TA 1535); 60 µg 2-AA (E. coli WP2 uvrA). Without S-9: 5 µg MNNG (TA 100, TA 1535), 10 µg NOPD (TA 98), 100 µg AAC (TA 1537), 10 µg ENNG (E. coli WP2 uvrA).

Details on test system and experimental conditions:

METHOD OF APPLICATION: standard plate test

DURATION
- Exposure duration: 48-72 h at 37 °C

NUMBER OF REPLICATIONS: each test was performed in triplicate

NUMBER OF COLONIES EVALUATED: all revertant colonies

DETERMINATION OF CYTOTOXICITY
- Method: titer determination; extent of background lawn


METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48-72 h

NUMBER OF REPLICATIONS: each test was performed in triplicate

NUMBER OF COLONIES EVALUATED: all revertant colonies

DETERMINATION OF CYTOTOXICITY
- Method: titer determination; extent of background lawn


GROWTH MEDIA:
S.Typhimurium: Vogel-Bonner E minimal glucose agar
E.Coli: SAI selective agar

Evaluation criteria:

In general, a substance to be characterized as positive in the bacterial tests has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

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:

cytotoxicity

Remarks:

A bacteriotoxic effect was observed at doses ≥ 2500 ug/plate (SPT) or ≥ 500 ug/plate (PIT).

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

≥ 2500 ug/plate (SPT) or ≥ 500 ug/plate (PIT).

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.