Disodium 4,4'-bis[[4-anilino-6-[(2-carbamoylethyl)(2-hydroxyethyl)amino]-1,3,5,-triazin-2-yl]amino]stilbene-2,2'-disulphonate

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: The test procedure cannot be subsumed under a testing guideline, nevertheless are well documented and scientifically acceptable. The test was not performed on GLP. The OECD recommended combination of strains was tested.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1980

Report date:

1980

Materials and methods

Principles of method if other than guideline:

The test was to be carried out for the purpose of checking the back mutation from Histidine requisite to non-requisite in Salmonella Tryphimurium and back mutation from Tryptophane requisite to non-requisite in Caliform Bacilli.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 4,4'-bis[[4-anilino-6-[(2-carbamoylethyl)(2-hydroxyethyl)amino]-1,3,5,-triazin-2-yl]amino]stilbene-2,2'-disulphonate

EC Number:

248-420-5

EC Name:

Disodium 4,4'-bis[[4-anilino-6-[(2-carbamoylethyl)(2-hydroxyethyl)amino]-1,3,5,-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Cas Number:

27344-06-5

Molecular formula:

C42H46N14O10S2.2Na

IUPAC Name:

disodium 4,4'-bis[[4-anilino-6-[(2-carbamoylethyl)(2-hydroxyethyl)amino]-1,3,5,-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: Histidine requisite strain.

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

other: Histidine requisite strain.

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

Additional strain / cell type characteristics:

other: Tryptophane requisite strain.

Metabolic activation:

with and without

Metabolic activation system:

S-9 additive

Test concentrations with justification for top dose:

5000, 1000, 500, 100, 50 and 10 µg/plate.

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

other: AF2, 4-nitro-o-phenylenediamine and 2-aminoanthracene.

Remarks:

- S9: TA98, TA 100 and WP2 uvrA AF2; TA 1537 9-AC, TA 1538 4-NOPD; TA 1535 ENNG. + S9: 2-AA all strains

Details on test system and experimental conditions:

PREPARATION OF SPECIMEN BACILLI
The specimen bacilli solution was made by supplementing 0.5 % NaCl, inoculating specimen bacilli into L-form test tube which contains nutrient broth (Difco), and cultivating them with two-way 16 hour shaking at the temperature of 37 °C.

TEST OPERATION
Plate Incorporation Method.
In the small tube, the specimen bacill solution 0.1 ml , specimen solution 0. 1 ml and phosphate-buffered solution 0.5 ml (S-mixed solution) 0.5ml for the metabolic activation test) were well mixed with 2 ml top agar and the mixture was put on the base agar plate to harden it.
The culture was to be made for 48 hours at 37 °C, and the generating mutation colony was counted.

REAGENTS
1) Top Agar
2) S-9 mixed solution
3) base agar

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

All six strains (Salmonella Typhinurium TA 1535, TA 100, TA 1537, TA 1538, TA 98; "Escherichia coll WP2 uvrA with the dose of 10 - 5000 µg both in the Direct and the Metabolic Activation Tests, remarkable increase in the mutation colony was not admitted.

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):
negative with and without metabolic activation

The test item is not judged to possess mutationality concerning the testings conducted with it.

Executive summary:

Method

The test item was evaluated for mutagenic potential in assays for gene mutation, without and with metabolic activation, in five tester strains of Salmonella thimurium and Escherichia coli WP2 avr A at the concentration of 5,000, 1,000, 500, 100, 50 and 10 µg/plate.

The test was to be carried out for the purpose of checking the back mutation from Histidine requisite to non-requisite in Salmonella Tryphimurium and back mutation from Tryptophane requisite to non-requisite in Caliform Bacilli.

Results

In all the strains tested (Salmonella Typhinurium TA 1535, TA 100, TA 1537, TA 1538, TA 98; Escherichia coll WP2 uv rA) with the dose of 10 - 5000 µg both in the Direct and the Metabolic Activation Tests, remarkable increase in the mutation colony was not admitted.

Conclusion

The test item is not judged to possess mutationality concerning the testings conducted with it.