Zirconium bis(hydrogen phosphate)

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:

2014-04-24 to 2014-05-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well documented, according to OECD guideline and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Zirconium bis(hydrogen phosphate)

EC Number:

237-401-7

EC Name:

Zirconium bis(hydrogen phosphate)

Cas Number:

13772-29-7

Molecular formula:

H3O4P.1/2Zr

IUPAC Name:

hydrogen phosphate;zirconium(4+)

Method

Target gene:

his trp

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:

Phenobarbital/β-naphthoflavone induced rat liver S9

Test concentrations with justification for top dose:

Pre-experiment/Experiment I: 3; 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate
Experiment II: 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate

Vehicle / solvent:

- Vehicle/solvent used: water
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties and its relative nontoxicity to the bacteria.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine, 2-aminoanthracene

Remarks:

With metabolic activation: 2-aminoanthracene (all strains), Without metabolic activation: sodium azide (TA 1535), 2-aminoanthracene , 4-nitro-o-phenylene-diamine(TA 1537, TA 98), methyl methane sulfonate (E. coli)

Details on test system and experimental conditions:

DETAILS ON TEST SYSTEM
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 37°C for 48 - 72 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
Toxicity detected by a
- decrease in the number of revertants
- clearing or diminution of the background lawn (= reduced his- or trp- background growth)
- reduction in the titer

Evaluation criteria:

A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.
A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.
An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.

Statistics:

According to the OECD guideline 471, a statistical analysis of the data is not mandatory.

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:

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:

Precipitation of the test item in the overlay agar on the incubated agar plates was observed from 1000 to 5000 µg/plate in both experiments with and without S9 mix. The undissolved particles had no influence on the data recording.

No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in the test groups with and without metabolic activation in both experiments

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Number of revertants per plate (mean of three plates), Experiment 1

	strain TA 1535		strain TA 1537		Strain TA 98		strain TA 100		E. coli WP2 uvrA
conc. [µg]	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA
deionized water	20	16	8	21	29	46	184	193	46	50
0	12	16	7	17	27	49	178	195	36	47
3	19	17	11	21	27	48	200	189	39	44
10	17	17	10	21	32	45	193	199	42	47
33	16	18	10	19	32	50	204	210	40	52
100	18	16	7	17	26	42	190	205	39	48
333	16	20	9	21	27	41	194	203	41	46
1000	15	14*	9*	24*	30*	46*	216*	197*	48*	38*
2500	15	18*	6*	22*	25*	39*	199*	196*	39*	46*
5000	11	12*	7*	11*	24*	37*	190*	209*	47*	56*
Sodium azide 10 µg	3339						1871
4-nitro-o-phenylene-diamine 10 µg					294
4-nitro-o-phenylene-diamine 50 µg			62
methyl methane sulfonate 2 µL									822
2-aminoanthracene 2.5 µg		619		371		3510		3403
2-aminoanthracene 10 µg										470

*Precipitate

 

Table 2: Number of revertants per plate (mean of three plates), Experiment 2

	strain TA 1535		strain TA 1537		Strain TA 98		strain TA 100		E. coli WP2 uvrA
conc. [µg]	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA
deionized water	17	24	10	18	29	44	176	199	40	53
0	12	18	11	17	25	33	193	206	44	41
10	14	22	10	22	27	41	205	217	39	47
33	19	21	9	19	27	41	209	205	35	52
100	19	24	9	9	29	46	180	232	40	55
333	15	19	9	21	27	42	199	198	43	43
1000	21*	15*	10*	15*	29*	40*	204*	215*	42*	53*
2500	8*	13*	8*	17*	29*	50*	189*	200*	44*	49*
5000	11*	13*	8*	14*	22*	34*	178*	210*	35*	53*
Sodium azide 10 µg	3206						1667
4-nitro-o-phenylene-diamine 10 µg					274
4-nitro-o-phenylene-diamine 50 µg			63
methyl methane sulfonate 2 µL									461
2-aminoanthracene 2.5 µg		531		316		3348		2735
2-aminoanthracene 10 µg										264

*Precipitate

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative