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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
Ames Test (BASF, 1986): negative (read across) Micronucleus Test (Liao et al., 1988): negative
Link to relevant study records
Reference
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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP, comparable to guideline study, available as unpublished report, no restrictions, fully adequate for assessment.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay
Target gene:
his
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9 (pretreated with Aroclor 1254)
Test concentrations with justification for top dose:
0, 20, 100, 500, 2500 and 5000 μg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (+S9); N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) for TA 100 and TA1535 (-S9), 4-nitro-o-phenylendiamine for TA98 (-S9), 9-aminoacridine chloride monohydrate for TA1537 (-S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) (tests 1 (TA 100 and TA98) and 2 (TA1535 and TA1537)) and preincubation (test 3)

DURATION
- Preincubation period: 20 minutes at 37 °C (test 3)
- Exposure duration: 48 hours at 37 °C in the dark (test 1, 2 and 3)

NUMBER OF REPLICATIONS: 3 tests were performed; 2 standard plate incorporation assays and 1 preincubation assay. In each test 3 test plates per dose were used.

DETERMINATION OF CYTOTOXICITY
- Method: reduced his(-) background growth
Evaluation criteria:
A substance is considered positive if it fulfills the following criteria:
1) doubling of the spontaneous mutation rate (control)
2) dose-response relationship
3) reproducibility of the results
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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY:
No bacteriotoxic effect (reduced his(-) background growth was observed.

TEST-SPECIFIC CONFOUNDING FACTORS:
Water solubility: complete solubility of the test substance in distilled water.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

General information

Water soluble zirconium salts like zirconium tetrachloride, zirconium diacetate or zirconium sulfate once in aqueous media dissociate very quickly at pH values of 4 to 9.5 forming the conjugate acid and hydrated forms of the insoluble ZrO2 (the most stable form of zirconium in water) (Daunderer, M.; Lehrbuch Klinische Toxikologie, 89. Erg. Lfg.; 11/94).

Due to the similar dissociation pattern it is concluded, that read across to other soluble zirconium salts is valid for the assessment of toxicological and ecotoxicological endpoints of zirconium(tetra)nitrate.

Bacterial reverse mutation assay- Ames

The read across substance zirconium diacetate (CAS# 5153-24-2) was tested for mutagenic effects in vitro in histidine-requiring strains of Salmonella typhimurium equivalent to OECD guideline 471 (BASF, 1986). The following strains were used: S. typhimurium TA 98, TA 100, TA 1535 and TA 1537. The test was performed with and without the addition of rat-liver post mitochondrial supernatant (S9 fraction) as an extrinsic metabolic activation system. The compound was dissolved in water and tested at five concentrations of 20, 100, 500, 2500, 5000 µg/plate in the presence and absence of a metabolic activation system. Each strain was additionally tested in the presence and in the absence of a metabolic activation system with a suitable known mutagen as positive control. The original experiment with and without metabolic activation was performed as standard plate incorporation assay. The confirmatory experiment with and without metabolic activation were carried out as preincubation assay.

In the experiments performed no reduced background lawn was observed. No genetic toxicity was observed in any of the tested strains. The read across substance is not genotoxic according to the results of this bacterial reverse mutation assay.

Bacterial reverse mutation assay- Fluctuation method

In a reverse gene mutation assay in bacteria, strains (TA97, TA100, TA102) of S. typhimurium were exposed to the read across substance zirconium tetrachloride (CAS# 10026-11-6) at concentrations of 0, 0.01, 0.1, 1.0 and 10.0 mg Zr/L in the absence of metabolic activation (Couture et al, 1989). Mutagenicity was assessed according to fluctuation method. It is performed entirely in liquid culture. The technique is the same as described for the traditional plate incorporation assay except that the whole assay is performed in 96-well micro plates with addition of a pH indicator. Metabolic processes of reproducing bacteria cause a drop in pH. The mutation frequency is counted as the number of wells out of 96 which have changed colour. Bromcresol Purple was used as pH indicator.

No genetic toxic effects were observed at any of the concentrations tested. The read across substance is not mutagenic according to the results of the bacterial reverse mutation assay.

Bacterial gene mutation assay-SOS Chromo assay

In a gene mutation assay in bacteria, a strain (PQ37) of E. coli was exposed to the read across substance zirconium tetrachloride (CAS# 10026-11-6) at concentrations ranging from 1.7 to 110 nM in the absence and presence of metabolic activation (Couture et al., 1989). Mutagenicity was assessed by a colorimetric assay. The expression of genes induced by genotoxic agents is measured by means of a fusion with the structural gene for beta-galactosidase. Activity of the protein is measured by adding lactose which yields a colored compound upon degradation. The quantifiable change in color is measured at 615 nm with a photometer.

No genetic toxic effects were observed at any of the concentrations tested with or without metabolic activation. The read across substance is not mutagenic according to the results of the bacterial reverse mutation assay.

The above results are further supported by an abstract of Liao et al. stating a negative result for the test substance zirconium(tetra)nitrate in a micronucleus assay (Liao et al., 1988).


Justification for selection of genetic toxicity endpoint
Most reliable study regarding the assessment of genotoxicity.

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available information is considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance does not need to be classified and labelled for mutagenicity under Directive 67/548/EEC, as amended for the 31st time in Directive 2009/2/EC.

 

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available information is reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance does not need to be classified and labelled for mutagenicity under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation EC No 605/2014.