Reaction mass of 6-Chloro-2-{4[ethyl(2-hydroxyethyl)amino]phenyl}-1-methylbenzo[cd]indolium hydroxide and 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate

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:

Study completion date: 18 November, 1983

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: As per the methodology proposed by Ames et al

Deviations:

not specified

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-bromo-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium hydroxide

IUPAC Name:

Reaction mass of 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-bromo-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium hydroxide

Test material form:

solid

Specific details on test material used for the study:

Name: FAT 31064/E
Purity: >95 %

Method

Target gene:

Histidine requiring genes of Salmonella strains

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

microsomal enzymes from rat liver

Test concentrations with justification for top dose:

0.2, 2, 20, 200 and 2000 µg/petri dish;

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: distilled water

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Without S9

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitrosoguanidine

Remarks:

Without S9

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: daunomycine

Remarks:

Without S9

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-anthramine

Remarks:

With S9

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: Triplicate
- Number of independent experiments : One

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: Background growth inhibition

METHODS FOR MEASUREMENTS OF GENOTOXICIY : Background growth inhibition

Evaluation criteria:

The criteria of mutagenicity used in this test are a doubling of the spontaneous reversion rate and a dose-effect relationship.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

other: Salmonella typhimurium TA 98 and TA 1537

Metabolic activation:

with

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

In the mutagenicity tests without and with metabolic activation, due to toxicity of the test material, a slight decline in the number of revertant colonies was occasionally observed with all strains at the highest concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

other: Salmonella typhimurium TA 98 and TA 1537

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

In the mutagenicity tests without and with metabolic activation, due to toxicity of the test material, a slight decline in the number of revertant colonies was occasionally observed with all strains at the highest concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

In the mutagenicity tests without and with metabolic activation, due to toxicity of the test material, a slight decline in the number of revertant colonies was occasionally observed with all strains at the highest concentrations.

Vehicle controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

No mutagenic effect was observed with the strains TA 1535 and TA 100.

The evidence for mutagenicity existed only in the presence of a liver microsomal enzyme preparation (S-9 mix) from male rats pretreated with Aroclor 1254 at the concentrations of 20 and 200 µg of product per Petri dish with TA 1537, and at 200 µg with TA 98. For these two strains a toxic effect was noted at the highest concentration of the product with a complete disappearance of the background lawn for TA 1537 and a dispersed lawn of auxotrophic bacteria for TA 98.

A toxic effect was also noted with strain TA 100 in the absence of metabolic activation at 2000 µg of product per Petri dish.

Applicant's summary and conclusion

Conclusions:

FAT 31064/F was found to exert a mutagenic action on strains S. typhimurium TA 98 and TA 1537 only in presence of metabolic activation.

Executive summary:

The mutagenic potential of FAT 31064/E was investigated in a bacterial reverse mutation assay according to procedure prescribed by Ames et al.

The substance FAT 31064/E was tested with the Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 at concentrations from 0.2 to 2000 µg (or nl) per petri dish - both in the presence and absence of metabolic activation.

A mutagenic effect was observed with strains TA 1537 and TA 98 only in presence of metabolic activation. No mutagenic effect was observed with strains TA 100 and TA 1535. A toxic effect was noted with strains TA 1537 and TA 100 at the highest tested concentrations.