1H-Benz[e]indolium, 2-[2-[2-chloro-3-[2-[1,3-dihydro-1,1-dimethyl-3-(4-sulfobutyl)-2H-benz[e]indol-2-ylidene]ethylidene]-1-cyclohexen-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfobutyl)-, inner salt, ammonium salt (1:1)

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:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-[(1E)-2-[(3Z)-2-chloro-3-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}cyclohex-1-en-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium amine

EC Number:

700-626-0

Cas Number:

1073607-50-7

Molecular formula:

C46 H50 Cl N2 O6 S2 . H4 N

IUPAC Name:

2-[(1E)-2-[(3Z)-2-chloro-3-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}cyclohex-1-en-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium amine

Method

Target gene:

histidine requiring gene (Salmonella typhimurium)
tryptophan requiring gene (Escherichia coli)

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9

Vehicle / solvent:

Dimethyl sulphoxide

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

dimethyl sulphoxide

Positive controls:

yes

Positive control substance:

other: TA1535: sodium azide, TA1537: ICR-191 (Acros Organics, Belgium), TA98: 2-nitrofluorene, TA100: methylmethanesulfonate, WP2uvrA: 4-nitroquinoline N-oxide

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

up to the top dose of 5000µg/plate (no increase revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

up to the top dose of 5000µg/plate (precipitation observed at 333µg/plate)(no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

up to the top dose of 5000µg/plate (no increase revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

up to the top dose of 5000µg/plate (precipitation observed at 333µg/plate)(no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

(no upper limit as no precipitation at 333µg/plate; increase observed but not biologically relevant increase of revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

(no upper limit as no precipitation at 333µg/plate; no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

(no upper limit as no precipitation at 333µg/plate; no increase of revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

(no upper limit as no precipitation at 333µg/plate; no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

up to precipitation at 1000µg/plate (no increase revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation at 1000µg/plate (no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 6

Species / strain:

S. typhimurium TA 100

Metabolic activation:

without

Genotoxicity:

negative

Remarks:

up to precipitation at 1000µg/plate (increase observed but not biologically relevant increase of revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation at 1000µg/plate (no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 7

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

negative

Remarks:

up to precipitation at 1000µg/plate (no increase revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation at 1000µg/plate (no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 8

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

up to precipitation at 1000µg/plate (no increase revertants)

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation at 1000µg/plate (no reduction bacterial background and no decrease revertants)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

The first and second above mentioned results are the dose range finding test, for which for the metabolic activation 5% S9 is used.
The third and fourth above mentioned results are the first mutation assay, for which for the metabolic activation 5% S9 is used.
The fifth above mentioned result is the second mutation assay, for which for the metabolic activation 5% S9 is used.
The sixth until last above mentioned results are the third mutation assay, for which for the metabolic activation 10% S9 is used.

Remarks on result:

other: strain/cell type: TA1537 and TA98

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Based on the results of this study it is concluded that the test substance is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.