REAKTIV OLIV F00-149

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:

2002-01-08 to 2002-04-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

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:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Guidelines for toxicity studies of drugs, 1990 MITI Guidelines for toxicity testing of chemicals

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

442-640-7

EC Name:

-

Cas Number:

479541-17-8

Molecular formula:

Hill formula: C32H26CuFN9Na4O19S5

IUPAC Name:

Tetrasodium 2-(4-fluoro-6-(2-(2-sulfonatoethansulfonyl)ethyl amino)-1,3,5-triazin-2-ylamino)-5-hydroxy-6-(2-(2-hydroxy-5- sulfonatophenylazo)-4,5-dimethoxyphenylazo)-7-sulfonatonaphthalene-1-sulfonatocuprate(II)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Reaktiv Oliv F00-0149

Method

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix from rat liver

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 50, 160, 500, 1600, and 5000 µg/plate
Concentration range in the main test (without metabolic activation): 50, 160, 500, 1600, and 5000 µg/plate

Vehicle / solvent:

Solvents:
deionized water for: sodium-azide,
DMSO for: 9-aminoacridine, 2-nitrofluorene, 4-nitroquinoline-N-oxide, 2-aminoanthracene

Controls

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

Remarks:

water only

True negative controls:

no

Positive controls:

yes

Remarks:

Without metabolic activation: sodium-azide for strain TA 100 and TA 1535, 9-aminoacridine for strain TA 1537, 2-nitrofluorene for strain T A 98, 4-nitroquinoline-N-oxide for strain WP2uvrA. With metabolic activation: 2-aminoanthracene for all strains

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: Pre-incubation involved incubating the test substance, S9-mix and bacteria for a short period of time before pouring this mixture onto plates of minimal agar.
- Exposure duration: 48h

NUMBER OF REPLICATIONS: 3 plates/strain/concentration

SELECTION AGENT (mutation assay): revert mutation

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: revert mutants

Evaluation criteria:

- Test substance produces at least a 2-fold increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control at complete bacterial background lawn.
- Test substance induces a dose-related increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control in at least two to three concentrations of the test compound at complete bacterial background lawn.

Statistics:

mean values reported

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

with metabolic activation

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

5000 µg/plate with strain T98 and TA1535

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'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive with metabolic activation
negative without metabolic activation

In this study, Reactive Olive F00-0149 is mutagenic in the presence of metabolic activation using the standard Ames Test procedure (plate incorporation test).

Executive summary:

In a reverse gene mutation assay in bacteria, strains TA 98, TA 100, TA 1535, TA 1537 of S. typhimurium and E. colli WP2 uvrA were exposed to Reactive Olive F00-0149 in deionized water at concentrations of 50, 160, 500, 1600, and 5000 µg/plate in the presence and absence of mammalian metabolic activation.

Reactive Olive F00-0149 was tested up to the limit concentration 5000 µg/plate.  The positive controls induced the appropriate responses in the corresponding strains.  There was evidence of a significant dose-response increase in the number of revertant colonies with metabolic activation with strain TA 98 at the highest dose and a slight increase in the number of reverant colonies with strain TA 1535 at the highest dose. 

This study is classified as acceptable and satisfies the requirement for Test Guideline  OPPTS 870.5100; OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.