Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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 guidelineopen allclose all
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
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

Constituent 1
Chemical structure
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 / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 98
Species / strain / cell type:
S. typhimurium TA 100
Species / strain / cell type:
S. typhimurium TA 1535
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
Controlsopen allclose all
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)
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
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 resultsopen allclose all
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
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
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
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.