Registration Dossier

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from publication.

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity Test: Test material
Author:
Mortelmans et al.
Year:
1986
Bibliographic source:
Environmental Mutagenesis

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Bacterial Reverse Mutation Assay was performed for the given test chemical.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- IUPAC Name: 4,4'-diaminostilbene-2,2'-disulphonic acid
- Mol. formula: C14H14N2O6S2
- Molecular Weight: 370.405 g/mole
- InChI: InChI=1S/C14H14N2O6S2/c15-11-5-3-9(13(7-11)23(17,18)19)1-2-10-4-6-12(16)8-14(10)24(20,21)22/h1-8H,15-16H2, (H,17,18,19)(H,20,21,22)/b2-1+
- Smiles: Nc1ccc(\C=C\c2ccc(N)cc2S(=O)(=O)O)c(c1)S(=O)(=O)O

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
not specified
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
Male Sprague-Dawley rats and male Syrian hamsters were routinely used for the preparation of the liver fractions
Test concentrations with justification for top dose:
0, 100.0, 333.0, 1000.0, 3333.0, 5000.0 ug/Plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: 95% Ethanol
- Justification for choice of solvent/vehicle: Test chemical was soluble in 95% Ethanol
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
95% Ethanol
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-Aminoanthracene (0.75 ug/Plate and 1.5 ug/Plate) and 4-Nitro-O-Phenylenediamine (12.0 ug/Plate)
Details on test system and experimental conditions:
METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): No data
- Test substance added in medium

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 37°C for 20 min
- Exposure duration/duration of treatment: 48 hr.
- Harvest time after the end of treatment (sampling/recovery times): No data
Evaluation criteria:
The criteria used for data evaluation were the same as those described previously [Haworth et al, 19831, and are summarized as follows: 1) mutagenic response: a dose-related, reproducible increase in the number of revertants over background, even if the increase was less than twofold; 2) nomutagenic response: when no increase in the number of revertants was elicited by the chemical; 3) questionable response: when there was an absence of a clear-cut dose-related increase in revertants; when the dose-related increases in the number of revertants were not reproducible; or when the response was of insufficient magnitude to support a determination of mutagenicity
Statistics:
No data

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA1535, TA1537, TA98, and TA100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
True negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES (if applicable): Chemical was initially tested with strain TA100 in the presence and the absence of the metabolic activation systems, over a wide dose range with an upper limit of 10 mgjplate, or less when solubility problems were encountered. Toxicity was evidenced by one or more of the following phenomena: appearance of hispinpoint colonies, reduced numbers of revertant colonies per plate, or thinning or absence of the bacterial lawn. Nontoxic chemicals were tested in the initial experiment up to the 10 mg/plate dose level, or to a level determined by their solubility. Toxic chemicals were tested up to a high dose which exhibited some degree of toxicity. As a rule, at least one toxic dose was incorporated into the first mutagenicity test; the repeat test(s) occasionally had the doses adjusted so that an apparent toxic dose was not reached.
Remarks on result:
other: No mutagenic potential

Applicant's summary and conclusion

Conclusions:
The given test chemical failed to induced mutation in Salmonella strains TA1535, TA1537, TA98, and TA100 in the presence and absence of S9 metabolic activation system and hence, it is not likely to be mutagenic.
Executive summary:

Bacterial Reverse Mutation Assay was performed for the given test chemical as per OECD Guideline 471 in S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 with or without S9 metabolic activation system extracted from Male Sprague-Dawley rats and male Syrian hamsters at concentrations of 0, 100.0, 333.0, 1000.0, 3333.0, 5000.0 ug/Plate. 95% Ethanol was used as a negative control. 2-Aminoanthracene, Sodium Azide, 9-Aminoacridine and 4-Nitro-O-Phenylenediamine were used as positive controls. Test was performed by preincubation assay. The criteria used for data evaluation were the same as those described previously [Haworth et al, 19831, and are summarized as follows: 1) mutagenic response: a dose-related, reproducible increase in the number of revertants over background, even if the increase was less than twofold; 2) nomutagenic response: when no increase in the number of revertants was elicited by the chemical; 3) questionable response: when there was an absence of a clear-cut dose-related increase in revertants; when the dose-related increases in the number of revertants were not reproducible; or when the response was of insufficient magnitude to support a determination of mutagenicity. The given test chemical failed to induced mutation in Salmonella strains TA1535, TA1537, TA98, and TA100 in the presence and absence of S9 metabolic activation system and hence, it is not likely to be mutagenic.