diethyleneglycyl bis (chloroacetate)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000-11-22 to 2000-11-30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

July 1997

Deviations:

yes

Remarks:

screening study with only one plate per dose

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-{2-[(2-chloroacetyl)oxy]ethoxy}ethyl 2-chloroacetate

Cas Number:

106-78-5

Molecular formula:

C8H12O5Cl2

IUPAC Name:

2-{2-[(2-chloroacetyl)oxy]ethoxy}ethyl 2-chloroacetate

Test material form:

liquid

Method

Target gene:

Salmonella typhimurium histidine system

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Remarks:

LT2

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

5000, 1600, 500, 160, 50, 16 µg/plate
5000 ug/plate is established in the standard protocol for use if not limited by solubility.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide, nitrofurantoin, 4-nitro-1,2-phenylene diamine, cumene hydroperoxide and 2-arminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation;
- Cell density at seeding (if applicable):

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours


NUMBER OF REPLICATIONS: one


DETERMINATION OF CYTOTOXICITY
- Method: background growth, reduction in the mutant count per plate, determination of titer

Evaluation criteria:

A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result For TA 1535, TA 100 and TA 98 this increase should be about twice that of negative controls, whereas for TA 1537, at least a threefold increase should be reached, For TA 102 an increase of about 100 mutants should be reached, Otherwise, the result is evaluated as negative.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

HISTORICAL CONTROL DATA
- Positive historical control data: yes
- Negative (solvent/vehicle) historical control data: yes

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- there was no indication of a bacteriotoxic effect of the substance at doses of up to and including 500 µg per plate. The total bacteria counts consistently produced results comparable to the negative controls, or differed only insignificantly. No inhibition of growth was noted as well. Higher doses had a strong, strain-specific bacteriotoxic effect. Therefore they could only partly be used for assessment purposes up to and including 1600 µg per plate.

Applicant's summary and conclusion

Conclusions:

The substance was considered to be non-mutagenic without and with S9 mix in the plate incorporation as well as in the preincubation screening.