Dilithium disodium (5,5'-diamino-(μ-4,4'-dihydroxy-1:2-κ-2,O4,O4',-3,3'-[3,3'-dihydroxy-1:2-κ-2-O3,O3'-biphenyl-4,4'-ylenebisazo-1:2-(N3,N4-η:N3',N4'-η)]-dinaphthalene-2,7-disulfonato(8)))dicuprate(2-)

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

Study period:

1990-04-25 to 1990-06-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted May 26, 1983

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Dilithium disodium (5,5'-diamino-(μ-4,4'-dihydroxy-1:2-κ-2,O4,O4',-3,3'-[3,3'-dihydroxy-1:2-κ-2-O3,O3'-biphenyl-4,4'-ylenebisazo-1:2-(N3,N4-η:N3',N4'-η)]-dinaphthalene-2,7-disulfonato(8)))dicuprate(2-)

EC Number:

407-230-4

EC Name:

Dilithium disodium (5,5'-diamino-(μ-4,4'-dihydroxy-1:2-κ-2,O4,O4',-3,3'-[3,3'-dihydroxy-1:2-κ-2-O3,O3'-biphenyl-4,4'-ylenebisazo-1:2-(N3,N4-η:N3',N4'-η)]-dinaphthalene-2,7-disulfonato(8)))dicuprate(2-)

Cas Number:

126637-70-5

Molecular formula:

Hill formula: C32 H16 Cu2 Li2 N6 Na2 O16 S4 CAS formula: C32 H16 Cu2 N6 O16 S4. 2Li 2Na

IUPAC Name:

dicopper(2+) dilithium(1+) disodium 4,4'-bis[2-(8-amino-1-oxido-3,6-disulfonatonaphthalen-2-yl)diazen-1-yl]-[1,1'-biphenyl]-3,3'-bis(olate)

Method

Target gene:

The strains are derived from S. typhimurium strain LT2 and due to a mutation in the histidine locus are histidine dependent.

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: Due to the "deep rough" (rfa -) mutation the bacteria possess a faulty lipopolysaccharide envelope which enables substances to penetrate the cell wall more easily. A further mutation causes a reduction in the activity of an excision repair system (uvrB -)

Metabolic activation:

with and without

Metabolic activation system:

Mammalian microsomal fraction S9 mix

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 10 ... 5000 µg/plate
Concentration range in the main test (without metabolic activation): 10 ... 5000 µg/plate

Vehicle / solvent:

Solvent: Distilled water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: Without metabolic activation: sodium azide (TA 1535, TA 100), 4-nitro-o-phenylene-diamine 4-NOPD (TA 1537, TA 98). With metabolic activcation: 2-aminoanthracene 2-AA (all strains)

Details on test system and experimental conditions:

METHOD OF APPLICATION:
This study was performed according to the plate incorporation test using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100. The assay was performed in two independent experiments, using identical procedures, both with and without liver microsomal activation. After solidification the plates were incubated for 72 hours at 37° C in the dark.

DURATION
- Preincubation: not applicable
- Exposure duration: 72 h
- Expression time: 72 h

NUMBER OF REPLICATIONS: 3 plates per concentration / control

DETERMINATION OF CYTOTOXICITY
- Method: Toxicity of the test article may be evidenced by a reduction in the number of spontaneous revertants, a clearing of the bacterial background lawn, or by degree of survival of treated cultures.

Evaluation criteria:

A test article is considered as positive if either a significant dose-related increase in the number of revertants or a significant and reproducible increase for at least one test concentration is induced.

A test article producing neither a significant dose-related increase in the number of revertants nor a significant and reproducible positive response at any one of the test points is considered non-mutagenic in this system.

A significant response is described as follows:
A test article is considered as mutagen if in strain TA 100 the number of reversions is at least twice as high and in strains TA 1535, TA 1537 and TA 98 it is at least three times higher as compared to the spontaneous reversion rate (defined in the original study report).
Also, a dose-dependent increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test article regardless whether the highest dose induced the above described enhancement factors or not.

Statistics:

No appropriate statistical method is available

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

1000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 1000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'. Remarks: experiment I and II

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The substance is considered to be non mutagenic in this Salmonella typhimurium reverse mutation assay.