Ammonium trioxovanadate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2006-09-05 till 2006-09-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented guideline study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

ammonium trivanadium octaoxide

IUPAC Name:

ammonium trivanadium octaoxide

Constituent 2

Reference substance name:

Ammonium trivanadium octaoxide

EC Number:

235-384-0

EC Name:

Ammonium trivanadium octaoxide

IUPAC Name:

235-384-0

Constituent 3

Reference substance name:

12207-63-5

Cas Number:

12207-63-5

IUPAC Name:

12207-63-5

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): "Ammoniumpolyvanadate (APV)"
- Molecular formula (if other than submission substance): (NH4)2V6O16
- Physical state: solid, yellow powder
- Storage condition of test material: at room temperature

Method

Target gene:

not applicable

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 97

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 98

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 100

Additional strain / cell type characteristics:

not specified

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 102

Additional strain / cell type characteristics:

not specified

Species / strain 5

Species / strain / cell type:

S. typhimurium TA 1535

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

Experiment I:
- without metabolic activation: 7, 21, 62, 185, 556 and 1667µg/plate
- with metabolic activation: 21, 62, 185, 556, 1667 and 5000µg/plate
Experiment II:
- without metabolic activation: 2.3, 7, 21, 62, 185 and 556µg/plate
- with metabolic activation: 7, 21, 62, 185, 556 and 1667µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: A 0.5% aqueous solution of Na-carboxymethylcellulose (CMC, Lot No. 98H0328, Sigma) was used as vehicle for the test substance and for the negative control group (vehicle control, 100µl).
- Justification for choice of solvent/vehicle: The test substance was not soluble in water or DMSO.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene; 1 or 2µg (dissolved in DMSO)

Remarks:

with metabolic activation; strains TA98, TA100 and TA1535

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

Remarks:

with metabolic activation; strain TA97a

Migrated to IUCLID6: ; 10µg (dissolved in DMSO)

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 1,8-Dihydroxy-anthraquinone; 50µg (dissolved in DMSO)

Remarks:

with metabolic activation; strain TA102

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

without metabolic activation; strain TA98

Migrated to IUCLID6: ; 2µg (dissolved in DMSO)

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

without metabolic activation; strains TA100 and TA1535

Migrated to IUCLID6: ; 1 or 2µg (dissolved in sterile water)

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-Nitro-o-phenylenediamine; 10µg (dissolved in DMSO)

Remarks:

without metabolic activation; strain TA97a

Controls 7

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: t-Butyl-hydroperoxide; 50µg (dissolved in sterile water)

Remarks:

without metabolic activation; strain TA102

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar
The test substance was tested with the plate incorporation method and the results were verified by a second independent experiment.

DURATION
- Exposure duration: 48 hours at 37°C

NUMBER OF REPLICATIONS: Triplicate repetitions were run for each dose group; for the control groups 6-fold repetitions were run.

COUNTING OF COLONIES: The plates with less than about 50 revertant colonies, i.e. the plates of TA98 and TA1535 with the exception of the positive controls, were counted visually. The other plates were photographed and the picture files were scanned for colonies by a computer program.

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth:
Different concentrations of test substance suspensions were mixed with phosphate buffer or S9 mix, bacteria (TA100) and top-agar and spread over a plate with minimal agar. The plates were incubated at 37°C for 48 hours and the growth of the bacterial background and the density of revertant colonies were determined.

OTHER EXAMINATIONS:
No further examinations.

Evaluation criteria:

A reproducible increase of the number of revertants to more than the following threshold values for at least one of the concentrations:
- For the strains with a low spontaneous revertant rate i.e. TA98 and TA1535: The 2.5-fold of the amount of the spontaneous revertants.
- For the strains with a high spontaneous revertant rate i.e. TA97a, TA100 and TA102: The 1.67-fold of the amount of the spontaneous revertants.

Statistics:

Means and standard deviations were calculated for the number of mutants in every concentration group.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

There was no increase in the number of mutants in any of the tested bacterial strains at any of the tested concentrations. The addition of an external metabolising system did not change these results.

Cytotoxicity / choice of top concentrations:

other: Slight toxicity to the bacteria was noted in the higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

There was no increase in the number of mutants in any of the tested bacterial strains at any of the tested concentrations. The addition of an external metabolising system did not change these results.

Cytotoxicity / choice of top concentrations:

other: Slight toxicity to the bacteria was noted in the higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

There was no increase in the number of mutants in any of the tested bacterial strains at any of the tested concentrations. The addition of an external metabolising system did not change these results.

Cytotoxicity / choice of top concentrations:

other: Slight toxicity to the bacteria was noted in the higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

There was no increase in the number of mutants in any of the tested bacterial strains at any of the tested concentrations. The addition of an external metabolising system did not change these results.

Cytotoxicity / choice of top concentrations:

other: Slight toxicity to the bacteria was noted in the higher concentrations.

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:

There was no increase in the number of mutants in any of the tested bacterial strains at any of the tested concentrations. The addition of an external metabolising system did not change these results.

Cytotoxicity / choice of top concentrations:

other: Slight toxicity to the bacteria was noted in the higher concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: pH 3.79 (1% suspension in deionised water, w/v, determined with a pH-Meter WTW pH 340)
- Water solubility: The test substance was not soluble.
- Precipitation: Test substance particles were visible in the higher concentration groups when the test substance was mixed with the agar.

RANGE-FINDING/SCREENING STUDIES: The test substance was toxic to the bacteria in the samples without metabolic activation at 5000 and 1667µg/plate and in the samples with metabolic activation at 5000µg/plate. Therefore 1667µg/plate was used as the highest concentration for the samples without metabolic activation and 5000 µg/plate as the highest concentration for the samples with metabolic activation which both could be toxic and 6 concentrations were tested each. Each of the other 5 concentrations was 1/3 of the preceding one.
The concentrations for the second experiment were changed due to the results of the first one: Toxicity was noted in the 1667 and 556µg/plate samples in the plates without metabolic activation and in the 5000 and 1667 µg/plate samples in the plates with metabolic activation. Therefore the concentrations were decreased one step each.

COMPARISON WITH HISTORICAL CONTROL DATA: Threshold values for evaluation of the results were derived from the variations in the control samples of available historical data of the Ames test.

ADDITIONAL INFORMATION ON CYTOTOXICITY: Slight toxicity to the bacteria was noted in the higher concentrations: Although the bacterial background was normal in the higher concentrations, the colonies were reduced in number and size, or no growth of bacteria was noted at all.

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):
negative

According to the results obtained in this study, the test substance "Ammoniumpolyvanadate (APV)" is non-mutagenic in the Ames test with the strains TA97a, TA98, TA100, TA102 and TA1535 with and without an external metabolic activation system up to the limit of toxicity.

Executive summary:

"Ammoniumpolyvanadate (APV)" was tested for mutagenic activity with the Ames test. The study was conducted in accordance with the OECD guideline 471. The test substance was suspended in 0.5% CMC. The following concentrations were tested: 7, 21, 62, 185, 556 and 1667µg/plate without metabolic activation and 21, 62, 185, 556, 1667 and 5000µg/plate with S9 mix. In a second experiment the concentrations were changed as follows: 2.3, 7, 21, 62, 185 and 556µg/plate without S9 mix and 7, 21, 62, 185, 556 and 1667µg/plate with S9 mix.

The test was performed according to the "direct plate incorporation method". The bacterial strains TA97a, TA98, TA100, TA102 and TA1535 were used. Negative and positive controls were included.

In the main test slight toxicity to the bacteria was noted in the higher concentrations: Allthough the bacterial background was normal in the higher concentrations, the colonies were reduced in number and size, or no growth of bacteria was noted at all.

In none of the concentrations tested and with none of the strains used an increase of the mutation frequency to more than the threshold values (250% of the controls for strains TA98 and TA1535, 167% of the controls for strains TA97a, TA100 and TA102) was obtained. Metabolic activation did not change these results.