Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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

Constituent 1
Chemical structure
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.