Ethyl chloroacetate

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:

From 11.01.1983 to 24-02-1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl chloroacetate

EC Number:

203-294-0

EC Name:

Ethyl chloroacetate

Cas Number:

105-39-5

Molecular formula:

C4H7ClO2

IUPAC Name:

ethyl 2-chloroacetate

Details on test material:

- Name of test material (as cited in study report): Monochloressigsäureethylester
- Physical state: colourless, clear
- Analytical purity: 99.5%
- Impurities (identity and concentrations): ca. 0.1 % Etnanol; ca. 0.1 % Water; ca. 0.3 % Dichloressigsäuremethylester (Methyl-2,2-dichloroacetate)
- Lot/batch No.: LKW Reitter August 2, 1982
- Storage condition of test material: dark at 2 ºC

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:

S-9 mix

Test concentrations with justification for top dose:

4, 20, 100, 500, 2500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: At the day of the experiment the test substance was dissolved in DMSO at appropriate concentrations.

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Remarks:

(DMSO (100 µL))

Remarks:

with and without metabolic activation

Controls 2

Positive controls:

yes

Remarks:

(without metabolic activation)

Positive control substance:

sodium azide

Remarks:

TA 100 and TA 1535; Sodium azide: 1 µg/plate

Controls 3

Positive controls:

yes

Remarks:

( without metabolic activation)

Positive control substance:

9-aminoacridine

Remarks:

TA 1537; 9-Aminiacridine: 50 µg/plate

Controls 4

Positive controls:

yes

Remarks:

(without metabolic activation)

Positive control substance:

2-nitrofluorene

Remarks:

TA 1538 and TA 98; 2-nitrofluorene: 5 µg/plate

Controls 5

Positive controls:

yes

Remarks:

(without metabolic activation)

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine

Remarks:

WP2uvrA; N-methyl-N'-nitro-N-nitrosoguanidine: 5 µg/plate

Controls 6

Positive controls:

yes

Remarks:

(with metabolic activation)

Positive control substance:

benzo(a)pyrene

Remarks:

10 µg/plate

Controls 7

Positive controls:

yes

Remarks:

(with metabolic activation)

Positive control substance:

other: 2-aminoanthracene

Remarks:

TA 100, TA 1538 and TA 98: 0.5 µg/plate; TA 1535 and TA 1537: 1 µg/plate; WP2uvrA: 10 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation).
Top agar is prepared for the Salmonella strains by mixing 100 mL agar (0,6 % agar, 0,5 % NaCl) with 10 ml of a 0,5 mM histidine-biotin solution. With
E. coli histidine is replaced by tryptophan (2,5 mL, 0,5 mM). The following ingredients are added (in order) to 2 mL of molten top agar at 45°C:
0,1 mL test compound solution
0,1 mL of an overnight nutrient broth culture of the bacterial tester strain
0,5 mL S-9 Mix (if required) or buffer
After mixing, the liquid is poured into a petridish with minimal agar (1,5 % agar, Vogel-Bonner E medium with 2 % glucose).

DURATION
- Exposure duration: 48-72 h

NUMBER OF REPLICATIONS: Triplicates

Evaluation criteria:

The test item is considered positive if there is a dose related increase in the number of revertants or a biologically relevant increase for at least one test concentration.

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:

(dose: 10000 µg/plate)

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:

cytotoxicity

Remarks:

(dose: 10000 µg/plate)

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:

cytotoxicity

Remarks:

(dose: 10000 µg/plate)

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
Preliminary toxicity tests were performed with all tester strains using a small number of plates to calculate an appropriate dose range. A reduced rate of spontaneously occuring colonies as well as visible thinning of the bacterial lawn were used as indicator for toxicity. Thinning of the bacterial lawn was controlled microscopically. The test compound was tested at doses of 4 to 10000 µg/plate and proved to be toxic to all bacterial strains at 10000 µg/plate. Thinning of the bacterial lawn and a reduction in the number of colonies has been observed at
10000 µg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: Control plates without mutagen showed that the number of spontaneous revertant colonies was similar to that described in the literature.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

In the absence of the metabolic activation system the test compound did not show a dose dependent influence in the number of revertants in any of the bacterial strains. Also in the presence of metabolic activation system, treatment of the cells with Monochloressigsäureethylester did not result in relevant increases in the number of revertant colonies. So it is considered that the test substance is not mutagenic in these bacterial test systems neither with nor without exogenous metabolic activation at the dose levels investigated.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Mutagenicity: In the absence of the metabolic activation system the test compound did not show a dose dependent influence in the number of revertants in any of the bacterial strains. Also in the presence of metabolic activation system, treatment of the cells with Monochloressigsäureethylester did not result in relevant increases in the number of revertant colonies. So it is considered that the test substance is not mutagenic in these bacterial test systems neither with nor without exogenous metabolic activation at the dose levels investigated.

Executive summary:

Monochloressigsäureethylester was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA. The mutagenicity studies were conducted in the absence and in the presence of a metabolizing system derived from rat liver homogenate. A dose range of 6 different doses from 4 µg/plate to 5000 µg/plate was used.

Control plates without mutagen showed that the number of spontaneous revertant colonies was similar to that described in the literature. All the positive control compounds gave the expected increase in the number of revertant colonies.

Toxicity: The test compound proved to be toxic to the bacteria at 5000 and 10000 µg/plate. 5000 µg/plate was chosen as top dose level for the mutagenicity study.

Mutagenicity: In the absence of the metabolic activation system the test compound did not show a dose dependent influence in the number of revertants in any of the bacterial strains. Also in the presence of metabolic activation system, treatment of the cells with Monochloressigsäureethylester did not result in relevant increases in the number of revertant colonies.

Summarizing, it can be stated that Monochloressigsäureethylester is not mutagenic in these bacterial test systems neither with nor without exogenous metabolic activation at the dose levels investigated.