Vanadyl pyrophosphate

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:

1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study conducted in accordance with internationally recognised test methods

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Vanadyl pyrophosphate

EC Number:

406-260-5

EC Name:

Vanadyl pyrophosphate

Cas Number:

58834-75-6

Molecular formula:

V2P2O9

IUPAC Name:

divanadium(4+) (phosphonatooxy)phosphonate dioxidandiide

Details on test material:

- Name of test material (as cited in study report): BTN/A
- Chemical name: Vanadyl pyrophosphate
- Chemical formula: (VO)2P207
- Relative molar mass: 307.8
- Physical state Fine, dark-brown powder
- Lot/batch No.:0003
- Purity: Vanadium - 30.1%, phosphorous 21.0% by weight
- Storage condition of test material: Ambient conditions, in the dark

Method

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:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat S-9 liver enzyme mix.

Test concentrations with justification for top dose:

25, 79, 250, 790, 2500 ug plate

Vehicle / solvent:

Distilled water

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene. Tested in the presence and absence of S-9 mix.

Remarks:

With strain TA1535

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

With strains TA100, TA1535

Migrated to IUCLID6: Tested only in the absence of S-9 mix

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

With strains TA98, TA1538

Migrated to IUCLID6: Tested only in the absence of S-9 mix.

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

With strains TA98, TA100, TA1537 and TA1538

Migrated to IUCLID6: Tested in the presence and absence of S-9 mix

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

With strain TA1537

Migrated to IUCLID6: Tested only in the absence of S-9 mix.

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Exposure duration: 2 days

SELECTION AGENT (mutation assays): Reversion from histidine dependence

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: other: thinnning of background lawn

Main assay conducted as follows:
An aliquot (0.1 ml) of each concentration of BTN/A was added to a sterile tube containing molten, histidine-deficient agar (2 ml) and bacterial suspension (0.1 ml) maintained at 45°C. Where appropriate, 0.5 ml rat liver microsomal preparation (S-9 mix) was added. The contents of the tubes were mixed thoroughly and poured onto plates containing solidified minimal medium (20 ml).
Further plates were prepared without the inclusion of the test organisms to verify the sterility of the S-9 mix and the test material. A control series of plates was prepared to confirm the inability of distilled water (0.1 ml) to induce reversion in the bacterial strains, and to provide a measure of the spontaneous mutation rates.
Aliquots (0.1 ml) of a 10-6 dilution of culture were spread on the surface of plates of complete medium to measure the viability and cell-density of each culture. All plates were prepared in triplicate, allowed to solidify and incubated at 37°C for 2 days. After incubation, numbers of revertant colonies were counted, either manually or with a Biotran II automatic colony counter. Total colonies on nutrient plates were counted in the same way. Growth of the background lawn of non-revertant cells on minimal plates was verified.

Evaluation criteria:

Sterility checks, spontaneous reversion rate and viability checks; activity of positive control agents all served to demonstrate correct functioning of the test system.
Action of BTN/A determined by changes in mutation frequency

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 2500 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 2500 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Sterility checks, spontaneousreversion rate and viability checks:
The absence of colonies on BTN/A and S-9 mix sterility check plates indicates that these preparations were free of significant microbial contamination.The total colony counts on untreated plates confirmed the viability of the cultures of the individual organisms.

Negative/vehicle controls:
The counts recorded on appropriate negative control plates confirmed the characteristically low spontaneous reversion rates of the tester strains and the absence of effects on these rates of incluson of the selected vehicle (distilled water inclusion).

Mutagenic activity of positive control chemicals
Appropriate positive control chemicals (with S-9 mix where required) induced marked increases in revertant colony numbers with all strains, confirming sensitivity of the cultures and activity of the S-9 mix.

Action of BTN/A
No increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to BTN/A at levels from 25 to 2500 ug per plate.Inhibition of growth, observed as slight thinning of the background lawn of non-revertant cells, occurred in all strains following exposure to BTN/A at 2500 ug per plate.

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

The test material, BTN/A, was devoid of mutagenic activity under the conditions of test.

Executive summary:

A bacterial reverse mutation assay (Ames test) has been undertaken following OECD/EU test methods.

The substance does not induce reverse mutation in Salmonella typhimurium.