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Toxicological information

Genetic toxicity: in vitro

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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

Constituent 1
Chemical structure
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 / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
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 resultsopen allclose all
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
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'.

Applicant's summary and conclusion