4,4'-ethylenedianiline

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well performed study following scientiffic priciples in compliance with GLP. A deviating set of tester strains as compared to actual guidelines was used.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Ames, B.N., Mc Cann, J. and Yamasaki, E., Mutation Res. (1975), 31, 347-364

Principles of method if other than guideline:

A deviating set of tester strains as compared to actual guidelines was used.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-ethylenedianiline

EC Number:

210-716-7

EC Name:

4,4'-ethylenedianiline

Cas Number:

621-95-4

Molecular formula:

C14H16N2

IUPAC Name:

4-[2-(4-aminophenyl)ethyl]aniline

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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254(R) induced rat liver S9 mix

Test concentrations with justification for top dose:

0, 1.58, 5, 15.8, 50, 158, 500, 1580 and 5000 µg / plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

methylmethanesulfonate

other: 2-Aminoanthracene

Details on test system and experimental conditions:

Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1,537 and TA 1538, obtained from B.N. Ames, University of California,
Berkeley CA, 947?0, USA, May 1?81.

The bacteria were grownn in a shaking water bath for 16 hours overnight at 37 degrees centigrade in 2.5% Nutrient Broth No. 2x..
After centrifugation, the bacteria were resuspended to a concentration of approximately 1 x 10 ep. 8 to 2 x 10 exp 9 cells per milliliter in 0.16 % Nutrient Broth and 0.5 % Sodium chloride. The concentration of germs was controlled photometrically and determined in an experimental test with
Histidine-rich potassium chloride solution on selective agar plates .

The plates were incubated at 37 degrees centigrads in the dark for 3 days.

Evaluation criteria:

A material is identified as a mutagen in this test system if there is a reproducible demonstration of a dose effect relation with a 2-foId increase in the number of revertants over the controls in a minimum of one strain.

With the strain TA 100 a 1.5-foId increase is the criterion for a positive result.

Statistics:

not required

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

except TA 1537

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

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

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive

In conclusion, it can be stated that during this in vitro SaImonella./mammalian-microsome assay and under the experimental conditions reported, the test item showed a reproducible dose-dependent mutagenic activity when tested with liver microsomal activation.

Executive summary:

This study was performed to investigate the mutagenic activity of the test item according to the plate incorporation test of Ames et. al . using the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, and TA 1538.

The test was performed with and without rat liver microsomal activation. The test material was tested at the following concentrations: 0, 1.58, 5, 15.8, 50, 158, 500, 1580 and 5000 micrograms per plate. Each concentrration, including the controls, was tested in triplicate.

No toxic effect of the test material was observed.

In the experiment without metabolic activation, no relevant increase of the revertant colony numbers was obtained in any SaImoneIla typhimurium strain, used at all dose levels tested when compared with the corresesponding controls.

In the experiment without metabolic activation no relevant increas of the revertant colony numbers was obtained in any tester strain. In the experiment with metabolic activation, a dose-dependent increase in the number of revertants was observed in all tester strains with the exception of TA 1537. A mutagenic activity of the test material was ascertained at the concentrations starting from 15.8 µg per plate. These findings were confirmed in an independent second experiment.