4,4'-diaminostilbene-2,2'-disulphonic acid

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:

1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Four strains were tested instead of 5.

Data source

Reference

Reference Type:

publication

Title:

No information

Author:

Zeiger E. et al.

Year:

1987

Bibliographic source:

Environ. Mutagen. 9, Suppl. 9, 1-110

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

not specified

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-diaminostilbene-2,2'-disulphonic acid

EC Number:

201-325-2

EC Name:

4,4'-diaminostilbene-2,2'-disulphonic acid

Cas Number:

81-11-8

Molecular formula:

C14H14N2O6S2

IUPAC Name:

2,2'-ethene-1,2-diylbis(5-aminobenzenesulfonic acid)

Details on test material:

Purity of the test material was 93.7%.

Method

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S-9 from Aroclor 1254-induced rat or hamster liver

Test concentrations with justification for top dose:

100, 333, 1000,
3333 or 5000 micrograms/ml

Vehicle / solvent:

ethanol

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

The positive controls in the absence of S-9 were sodium azide (TA1535 and TA100), 9-aminoacridine (TA97 and TA1537), and 4-nitro-o-phenylenediamine (TA98). The positive control in the presence of S-9 was 2-aminoanthracene for all strains.

Positive control substance:

other: The positive controls in the absence of S-9 were sodium azide (TA1535 and TA100), 9-aminoacridine (TA97 and TA1537), and 4-nitro-o-phenylenediamine (TA98). The positive control in the presence of S-9 was 2-aminoanthracene for all strains.

Details on test system and experimental conditions:

Test material was dissolved in 95% ethanol and incubated with S. typhimurium strains TA98, TA100, TA1535 and TA1537 at 100, 333, 1000,
3333 or 5000 micrograms/ml with or without S-9 from Aroclor 1254-induced rat or hamster liver (10%) for 20 min at 37 degrees C without shaking. Top agar was added and the contents of the tubes were mixed and poured onto the surfaces of petri dishes that contained Vogel-Bonner medium. The colonies present after 2 days of incubation at 37 degrees were handcounted if there was a precipitate; otherwise automatic colony counters were used. Tests were performed in triplicate.

Evaluation criteria:

A chemical was judged to be a mutagen if a dose-related increase over the solvent control was observed and weakly mutagenic if a low-level dose
response was seen in duplicate tests. A test was considered questionable if the number of mutants at a single dose was elevated or if an increase that was not dose-related was seen.

Statistics:

no data

Results and discussion

Test results

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:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative.

Executive summary:

The potential for the test substance to induce an increase in reverse mutations in bacteria cells was evaluated in an AMES tes, according to the OECD Guideline 471. None of the concentrations tested caused an increase in the number of mutants in the absence or presence of 10% rat or hamster liver S-9. All positive controls induced at least a 2-fold increase in the number of mutants with respect to the vehicle control.