3,4-dichloroaniline

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: Comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1984

Report date:

1984

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

yes

Remarks:

; no strain with AT-baise pair as primary reverse mutation site was used

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

; no strain with AT-baise pair as primary reverse mutation site was used

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3,4-dichloroaniline

EC Number:

202-448-4

EC Name:

3,4-dichloroaniline

Cas Number:

95-76-1

Molecular formula:

C6H5Cl2N

IUPAC Name:

3,4-dichloroaniline

Details on test material:

- Name of test material (as cited in study report): 3,4-dichloroanilin
- Analytical purity: 100 % (GC)

- Lot/batch No.: Pt. 3382

- Stability under test conditions: tested and approved

Method

Target gene:

no data

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 98

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 100

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 1537

Metabolic activation:

with and without

Metabolic activation system:

Liver S9-Mix of Aroclor 1254 induced male Sprague Dawley rats

Test concentrations with justification for top dose:

test 1: 20, 100, 500, 2500, 12500 µg/plate
test 2: 62.5, 125, 250, 500, 1000, 2000 µg/plate

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

deionised water

True negative controls:

no

Positive controls:

yes

Remarks:

used for TA 1535 and TA 100

Positive control substance:

cyclophosphamide

Remarks:

Migrated to IUCLID6: 100 µg/plate (TA1535); 200 µg/plate (TA100)

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Remarks:

used for TA 1537 and TA 98

Positive control substance:

other: 50 µg/plate trypaflavine

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Remarks:

TA 1535, TA 1537, TA 100, TA 98

Positive control substance:

other: 3 µg/plate 2-aminoanthracen

Details on test system and experimental conditions:

according to Ames et al. (1973, 1975)
With and without metabolic activation 4 plates per tester strain and dose were used. After 48 h of incubation on a histidine deficient agar at 37°C the colonies were counted.

Evaluation criteria:

a reproducible, dose-dependent dublication of mutant counts in treated plates versus control plates in at least one strain was judged a positive result.

Statistics:

no data

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

at doses up to 500 µg/plate

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at doses of 1000 µg/plate or higher

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

at doses up to 250 (test 2) to 500 (test 1) µg/plate

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at doses of 500 (test 2) to 1000 (test 1) µg/plate or higher

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

at doses up to 500 µg/plate

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at doses of 1000 µg/plate or higher

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

at doses up to 100 (test 1) to 500 (test 2) µg/plate

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at doses of 500 (test 1) to 1000 (test 2) µg/plate or higher

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: strain/cell type: S. typhimurium TA 98

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

no further data

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Executive summary:

Herbold, B, Bayer-report no. 13085, 1984

3,4 -DCA was tested for mutagenicity in Salmonella typhimurium ( TA98, TA100, TA 1535 and TA 1537) similar to OECD 471 and EU methods B.13/14 (without testing of A/T base pair revertant strains). The tests were conducted according to the method of Ames et al. (1975). After 48 h of incubation growth of revertant colonies on histidine deficient selection agar was counted and compared to vehicle controls. 3,4 dichloroaniline did not produces a reproducible, dose related response over the solvent control, under a single metabolic activation condition, in replicate trials. The obtained results therefore indicate that 3,4 -DCA in a concentration of up to 500 µg/ plate appeared to be nonmutagenic in these Salmonella strains. Higher doses were cytotoxic, as indicated by reduced His+ revertant colonies.