Tricopper bis(orthophosphate)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 Nov 2010 - 17 Dec 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

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:

Dicopper hydroxide phosphate

EC Number:

235-285-2

EC Name:

Dicopper hydroxide phosphate

Cas Number:

12158-74-6

Molecular formula:

Cu2HO5P

IUPAC Name:

copper(2+) hydroxide phosphate (2:1:1)

Method

Target gene:

Histidine for Salmonella.

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 97a, TA 98, TA 100, TA 102, TA 1535

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

Conc 1: 50 mg/mL
Conc 2: 15.82 mg/mL
Conc 3: 5.01 mg/mL
Conc 4: 1.58 mg/mL
Conc 5: 0.5 mg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: phosphate buffered saline
- Justification for choice of solvent/vehicle: no data

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

sodium azide

benzo(a)pyrene

mitomycin C

other: Daunomycin, ICR 191

Remarks:

See table 1 for details on the use of positive controls

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 minutes at 37°C
- Exposure duration: incubation period was 48 hours.

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA 97a, TA 98, TA 100, TA 102, TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

The results of the negative and positive controls confirm the sensitivity and accuracy of the test system, according to the requirements of the Moltox-kit. In general, the positive control frequencies should be at least 2.5 times the negative counts (spontaneous frequency).

Any other information on results incl. tables

RESULTS

Table 2: Experiments with test strain TA 97a

Control mutagen: ICR 191 acridine

Experiment without S9-mix		Experiment with S9-mix
Mutagen µg/ml	Number of revertants	Mutagen µg/mL	Number of revertants
10	1191	100	493
Spontaneous revertants:131		Spontaneous revertants:127
Test preparation concentration	Number of revertants	Test preparation concentration	Number of revertants
50 mg/mL	94	50 mg/mL	80
15.82 mg/mL	127	15.82 mg/mL	93
5.01 mg/mL	132	5.01 mg/mL	88
1.58 mg/mL	93	1.58 mg/mL	109
0.5 mg/mL	68	0.5 mg/mL	76

 

No genotoxic activity observed.

Table 3: Experiments with test strain TA 98

Control mutagen: Daunomycin

Experiment without S9-mix		Experiment with S9-mix
Mutagen µg/ml	Number of revertants	Mutagen µg/mL	Number of revertants
60	555	100	338
Spontaneous revertants:21		Spontaneous revertants: 21
Test preparation concentration	Number of revertants	Test preparation concentration	Number of revertants
50 mg/mL	31	50 mg/mL	33
15.82 mg/mL	27	15.82 mg/mL	25
5.01 mg/mL	27	5.01 mg/mL	21
1.58 mg/mL	15	1.58 mg/mL	25
0.5 mg/mL	16	0.5 mg/mL	20

No genotoxic activity observed.

 

Table 4: Experiments with test strain TA 100

Control mutagen: Sodium azide

Experiment without S9-mix		Experiment with S9-mix
Mutagen µg/ml	Number of revertants	Mutagen µg/mL	Number of revertants
15	>300	100	>300
Spontaneous revertants:65		Spontaneous revertants:79
Test preparation concentration	Number of revertants	Test preparation concentration	Number of revertants
50 mg/ml	63	50 mg/mL	68
15.82 mg/ml	50	15.82 mg/mL	60
5.01 mg/ml	36	5.01 mg/mL	41
1.58 mg/ml	31	1.58 mg/mL	44
0.5 mg/ml	17	0.5 mg/mL	47

No genotoxic activity observed.

 

Table 5: Experiments with test strain TA 102

Control mutagen: Mytomycin C

Experiment without S9-mix		Experiment with S9-mix
Mutagen µg/ml	Number of revertants	Mutagen µg/mL	Number of revertants
5	754	100	874
Spontaneous revertants:221		Spontaneous revertants:277
Test preparation concentration	Number of revertants	Test preparation concentration	Number of revertants
50 mg/ml	213	50 mg/mL	216
15.82 mg/ml	192	15.82 mg/mL	250
5.01 mg/ml	211	5.01 mg/mL	256
1.58 mg/ml	205	1.58 mg/mL	272
0.5 mg/ml	145	0.5 mg/mL	217

No genotoxic activity observed.

 

Table 6: Experiments with test strain TA 1535

Control mutagen: sodium azide

Experiment without S9-mix		Experiment with S9-mix
Mutagen µg/ml	Number of revertants	Mutagen µg/mL	Number of revertants
15	339	100	381
Spontaneous revertants:7		Spontaneous revertants:8
Test preparation concentration	Number of revertants	Test preparation concentration	Number of revertants
50 mg/ml	10	50 mg/mL	7
15.82 mg/ml	7	15.82 mg/mL	6
5.01 mg/ml	7	5.01 mg/mL	6
1.58 mg/ml	7	1.58 mg/mL	10
0.5 mg/ml	4	0.5 mg/mL	6

 

No genotoxic activity observed.

Applicant's summary and conclusion

Conclusions:

The test material was determined to be non-genotoxic under the conditions of the study.