Dihydrogen (ethyl)[4-[4-[ethyl(3-sulphonatobenzyl)]amino]-2'-sulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, disodium salt

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:

2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Dihydrogen (ethyl)[4-[4-[ethyl(3-sulphonatobenzyl)]amino]-2'-sulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, disodium salt

EC Number:

223-339-8

EC Name:

Dihydrogen (ethyl)[4-[4-[ethyl(3-sulphonatobenzyl)]amino]-2'-sulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, disodium salt

Cas Number:

3844-45-9

Molecular formula:

C37 H34 N2 Na2 O9 S3

IUPAC Name:

disodium 2-({4-[ethyl(3-sulfonatobenzyl)amino]phenyl}{4-[ethyl(3-sulfonatobenzyl)iminio]cyclohexa-2,5-dien-1-ylidene}methyl)benzenesulfonate

Test material form:

solid: particulate/powder

Details on test material:

- appearance: blue powder
- storage conditios: at room temperature

Specific details on test material used for the study:

-Name in report: PURICOLOR BLEU ABL9
- Purity: 93%
- Other name: VIBRACOLOR Blue ABL9
-Batch no: 001641D0A
-appearance: blue powder
-date of receipt: 15 May 2003
-storage conditios: at room temperature

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

liver microsomal fraction (S9 fraction) of rats induced with Aroclor 1254.

Test concentrations with justification for top dose:

Pretest: 10, 100, 500, 1000, 2500 and 5000 μg/plate.
31.3, 62.5, 125, 250 and 500 μg/plate: for both experiments with S9 mix as well as for the first and second experiment without S9 mix,
125, 200, 250, 375, 500 and 625 μg/plate, for the third confirmatory experiment without S9 mix (only TA 1537)

Vehicle / solvent:

water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-Anthramine

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) with the exception of the second experiment with S9-mix (preincubation)

Evaluation criteria:

A reproducible 2-fold increase (for the TA 98, TA 100, TA 102 and WP2 uvrA sfrains) or 3-fold increase (for the TA 1535 and TA 1537 sfrains) in the number of revertants compared with the vehicle confrols, in any sfrain at any dose-level and/or evidence of a dose-relationship was
considered as a positive result. Reference to historical data, or other considerations of biological relevance may also be taken into account in the evaluation of the data obtained.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: Strong coloration by the test substance made the plates unreadable at doses of 1 mg/plate and above

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

Cytotoxicity / choice of top concentrations:

other: strong coloration of the test substance

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

A moderate to marked coloration was observed in the Petri plates when scoring the revertants at all dose-levels. This coloration made the plates unreadable at dose-levels > 1 mg/plate.
No noteworthy toxicity was noted towards the strains used, with and without S9 mix. No precipitate was observed in the Petri plates when scoring the revertants at all dose-levels.

No precipitate was observed in the Petri plates when scoring the revertants at all dose-levels.

Applicant's summary and conclusion

Conclusions:

CAS 3844-45-9 is not mutagenic in bacteria.