Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

An AMES test is available performed with bismuth oxy-iodide bromide according to OECD/ EC guidelines and GLP principles, with negative result. No other data on genotoxic properties of the target chemical are available, but other mode-of-actions are addressed by data from substance analogue bismuth vanadate (negative in an in vitro chromosome aberration test), and data on individual ions (bromide was shown to be negative in an in vitro chromosome aberration test and an in vitro UDS test, iodide was negative in an in vitro COMET assay and an in vitro mammalian cell gene mutation test). The endpoint conclusion is drawn following a weight-of-evidence approach.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
05 July - 09 Aug 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Target gene:
- S. typhimurium: Histidine gene
- E. coli: Tryptophan gene
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone
Test concentrations with justification for top dose:
Experiment 1
Preliminary test (without and with S9) TA100 and WP2uvrA: 3, 10, 33, 100, 333, 1000, 3330 and 5000 µg/plate
Main study: TA1535, TA1537 and TA98:
Without and with S9-mix: 33, 100, 333, 1000 and 3330 µg/plate
Experiment 2:
Without and with S9-mix: 33, 100, 333, 1000, 3330 and 5000 µg/plate
Vehicle / solvent:
- Vehicle used: DMSO
- Justification for choice of vehicle:
A homogeneous suspension could be in DMSO and DMSO is accepted and approved by authorities and international guidelines.
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Remarks:
TA1535: sodium azide 5 μg/plate; TA1537: 9-aminoacridine 60 μg/plate; TA98 2-nitrofluorene 10 μg/plate; TA100: methylmethanesulfonate 650 μg/ plate; WP2uvrA: 4-nitroquinoline N-oxide 10 μg/ plate.
Remarks:
Without metabolic activation
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Remarks:
2-aminoanthracene (2AA) at 1 μg (TA98 and TA100), 2.5 μg/ plate (TA1535, TA1537) and 10 μg (WP2uvrA).
Remarks:
With metabolic activation (5% S9-mix)
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Remarks:
2-aminoanthracene (2AA) at 1 μg (TA98, TA100), 2.5 μg/ plate (TA1535), 5 μg (TA1537) and 10 μg (WP2uvrA).
Remarks:
With metabolic activation (10% S9-mix)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hour

NUMBER OF REPLICATIONS:
- Doses of the test substance were tested in triplicate in each strain. Two independent experiments were conducted.

NUMBER OF CELLS EVALUATED: 10E8 per plate

DETERMINATION OF CYTOTOXICITY
- Method: The reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies.

OTHER EXAMINATIONS:
- The presence of precipitation of the test compound on the plates was determined.
Evaluation criteria:
A test substance is considered negative (not mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 is not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537, TA98 or WP2uvrA is not greater than three (3) times the concurrent control.
b) The negative response should be reproducible in at least one independently repeated experiment.

A test substance is considered positive if:
a) A two-fold (TA100) or more or a three-fold (TA1535, TA1537, TA98, WP2uvrA) or more increase above solvent control in the mean number of revertant colonies is observed in the test substance group.
b) In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one independently repeated experiment.
Key result
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, but tested up to precipitating concentrations
Remarks:
Precipitation at 3330 μg/ plate and above.
Vehicle controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Remarks:
Precipitation at 3330 μg/ plate and above.
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Slight precipitation was observed at dose levels of 3330 and 5000 µg/plate.

RANGE-FINDING/SCREENING STUDIES:
- No toxicity or mutagenicity was observed up to and including the top dose of 5000 µg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA:
- The negative and strain-specific positive control values were within laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- No toxicity or mutagenicity was observed up to and including the top dose of 5000 µg/plate.
Conclusions:
In an AMES test, performed according to OECD/ EC guidelines and GLP principles, bismuth oxy-iodide bromide was found not to be mutagenic with or without metabolic activation.
Executive summary:

An AMES test was performed according to OECD/ EC guidelines and GLP principles. All bacterial strains showed negative responses up to and including 5000 ug/plate, i.e. no significant dose-related increase in the number of revertants with or without metabolic activation was seen. No cytotoxicity of the test substance was observed, but the test substance precipitate at 3330 ug/plate and above. The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Based on the results of this study it is concluded that bismuth oxy-iodide bromide is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay with or without metabolic activation.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
The rationale to read across the data is attached in section 13.
Reason / purpose:
read-across source
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
In every test under all concentration range, the test substance did not induce any chromosome aberration, i.e. polyploidy and structural (see Table2)
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
see Table 1
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid

Table 1: Cell growth index

 

Without metabolic activation

With metabolic activation

 

Concen-tration (µg/ml)

Cell growth index (%)

Frequency of mitotic cells

Frequency of aberration cells (50 cells)

 

Concentra-tion (µg/ml)

Cell growth index (%)

Frequency of mitotic cells

Frequency of aberration cells (50 cells)

Polyploid

Structural

Polyploid

Structural

 

 

 

24 hours treatment

0

100

++

0

0

 

 

 

+ S9 mix

0

100

++

0

0

39.1*

92.1

++

2

0

625*

91.9

++

2

0

78.1*

90.5

++

0

0

1250*

93.8

++

4

0

156*

75.5

++

0

0

2500*

97.5

++

2

2

313*

67.6

++

0

0

5000*

92.5

++

4

0

625*

51.9

++

0

0

X

X

X

X

X

1250*

41.5

+

2

0

X

X

X

X

X

2500*

39.8

±

X

X

X

X

X

X

X

5000*

46.9

-

X

X

X

X

X

X

X

48 hours treatment

0

100

++

0

0

 

 

-S9 mix

0

100

++

0

0

156*

93.3

++

0

0

625*

85.4

++

0

0

313*

84.2

++

0

0

1250*

81.7

++

0

0

625*

62.3

++

0

0

2500*

82.3

++

4

0

1250*

34.4

+

4

0

5000*

36.6

++

0

2

2500*

23.2

-

X

X

X

X

X

X

X

5000*

25.2

-

X

X

X

X

X

X

X

*Average of 2 plates; ++Many mitotic cells; +Small amount of mitotic cells; ±Very small amount of mitotic cells; -No mitotic cells

Table 2: Results of chromosomal aberration test (without metabolic activation)

 

Treatment

Treatment time (h)

Concentration (µl/ml)

Total observe (100+100)

Total number of polyploids

Judg-ment

Number and percentage (%) of cells showing chromosomal aberrarions

Gap

Chromatid-type

Chromodome-type

Others

Total

Judg-ment

g

ctb

cte

csb

cse

-g

+g

Solvent (0.5% CMC)

24

0

200

1 (0.5)

 

0 (0.0)

0 (0.0)

1 (0.5)

0 (0.0)

0 (0.0)

0 (0.0)

1 (0.5)

1 (0.5)

 

48

0

200

0 (0.0)

 

0 (0.0)

0 (0.0)

1 (0.5)

0 (0.0)

1 (0.5)

0 (0.0)

2 (1.0)

2 (1.0)

 

 

 

Test substance

 

24

313*

200

1 (0.5)

-

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

-

625*

200

2 (1.0)

-

0 (0.0)

0 (0.0)

1 (0.5)

0 (0.0)

0 (0.0)

0 (0.0)

1 (0.5)

1 (0.5)

-

1250*

200

5 (2.5)

-

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

1 (0.5)

0 (0.0)

1 (0.5)

1 (0.5)

-

 

48

313*

200

2 (1.0)

-

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

1 (0.5)

0 (0.0)

1 (0.5)

1 (0.5)

-

625*

200

4 (2.0)

-

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

-

1250*

200

5 (2.5)

-

0 (0.0)

0 (0.0)

0 (0.0)

0 (0.0)

1 (0.5)

0 (0.0)

1 (0.5)

1 (0.5)

-

Positive control (MMC)

24

0.05

200

1 (0.5)

-

2 (1.0)

17 (8.5)

54 (27.0)

0 (0.0)

1 (0.5)

0 (0.0)

69(34.5)

71(35.5)

++

48

0.05

200

3 (1.5)

-

7 (3.5)

16 (8.0)

98 (49.0)

2 (1.0)

7 (3.5)

0 (0.0)

114(57.0)

119(59.5)

+++

Fill in the number of cells observed, polyploids and cells showing total structural chromosomal aberrations in 2 plates, and the percentage (%) of cells showing aberrations in parenthesis with every concentration of treatment. Gap (g) – Fill in the total value of chromatd-type and chromosome-type. Sum total (-g) – Fill in the total number and percentage of cells except those which has only gaps. If there are many aberrations in one cell, calculate that cell as one cell showing aberrations. For example, if there are 2 exchanges in one cell, calculate that cell as one showing aberrations. One cell showing break and one cell showing exchange. ctb: chomatid break; cte: chromatid exchange; cse: chromosome exchange; others: fragmentation etc. (except pulverization)

Conclusions:
In a chromosome aberration test, performed equivalent to OECD guideline 473, BiVO4 did not show genotoxicity when tested up to 5000 ug/ plate. This result is read across to bismuth oxy-iodie bromide.
Endpoint:
in vitro cytogenicity / micronucleus study
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
The rationale to read across the data is attached in section 13.
Reason / purpose:
read-across source
Key result
Species / strain:
Chinese hamster Ovary (CHO)
Remarks:
CHO-K1
Metabolic activation:
not applicable
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:
not examined
Positive controls validity:
valid
Conclusions:
The read-across analogue test substance potassium iodide did not induce DNA damage in the in vitro alkaline Comet assay up to the concentration of 10 mM. This conclusion can be read across to bismuth oxy-iodide bromide.
Executive summary:

In an in vitro alkaline Comet assay, the test substance potassium iodide did not induce DNA damage up to and including the concentration of 10 mM. There was no cytotoxicity. Exposure duration was 3 hours and cells were incubated in the medium for 0.5, 1 or 24 hours. Two independent assays were performed, and each concentration was tested in duplicate. The positive control etoposide, a well known inhibitor of topoisomerase II inducing DNA double-strand breaks, produced satisfactory positive response. Based on the results, the test substance potassium iodide is considered to be not genotoxic under conditions of the study.

These results can be read across to bismuth oxy-iodide bromide.
Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
The rationale to read across the data is attached in section 13.
Reason / purpose:
read-across source
Key result
Species / strain:
lymphocytes:
Remarks:
human
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:
not applicable
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: No significant alterations in mitotic indices were found at any concentraton up to 5000 µg/mL in the preliminary range-finding test. In the absence of activation, mean percent of dividing (metaphase) cells ranged from 7.8 to 11.7 with no dose-relationship, compared to the water (solvent) control mean of 9.8%, while in S9-supplemented cultures the test range was again a flat 7.8 to 11.6% compared to the control value of 7.7%.

Remarks on result:
other: negative
Conclusions:
The read-across analogue substance sodium bromide was not cytogenic in a chromosome aberration test with human lymphocytes, both with and without metabolic activation, up to the limit dose of 5000 µg/mL. These results can be read-across to bismuth oxy-iodide bromide.
Executive summary:

In a GLP-compliant chromosome aberration study performed according to OECD guideline 473, a read-across analogue chemical sodium bromide was tested up to the concentration of 5000 µg/mL both in the absence and presence of metabolic activation. The exposure duration was 2 hours in the presence and 22 hours in the absence of metablic activation. No cytotoxicity was observed at any concentration level. The microscope preparations of colchicine-treated cells were scored for chromosome aberrations (100 metaphases/culture). The test substance did not induce any significant increases either in the proportion of metaphases with chromosome aberrations, or the type of aberrations. Positive controls produce satisfactory response. Based on these results, sodium bromide showed no evidence of clastogenicity under conditions of the study. These results can be read across to bismuth bromide iodide oxide.

Endpoint:
in vitro DNA damage and/or repair study
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
The rationale to read across the data is attached in section 13.
Reason / purpose:
read-across source
Key result
Species / strain:
mammalian cell line, other: human HeLa-S3
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: In both tests, the HDT of the test compound caused some toxicity, but at no test concentration was there any consistent increase in grain count over solvent control.

In the second activated test, a small bu significant increase was recorded in gross nuclear count at a single concentration, 1600 µg/mL; however, since the cytoplasmic (background) labeling level was comarably high, there was no increase in the corresponding net nuclear grain count. Another isolated small but significant increase in net nuclear grain count was registered in this second activated test at 400 µg/mL, although no corresponding increase was recorded in the gross nuclear grain count. Both these increases were discounted as representing chance variations, since they were unrelated to dose and not reproducible between tests.

Conclusions:
In a study performed according to OECD Guideline 482, the read-across analogue test substance sodium bromide showed no evidence of DNA damage in human HeLa-S3 cell line, both with and without metabolic activation, as measured by silver grain counts (unscheduled DNA synthesis). These results can be read across to bismuth oxy-iodide bromide.
Executive summary:

In a GLP-compliant study performed according to OECD guideline 482, the read-across analogue test substance sodium bromide did not cause DNA damage in human HeLa-S3 cell line, up to the maximal test concentration of 25600 µg/mL, both with and without metabolic activation, as measured by silver grain counts (unscheduled DNA synthesis). The exposure duration was 3 hours. and the experiments were performed in duplicate. These results can be read across to bismuth oxy-iodide bromide.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
The rationale to read across the data is attached in section 13.
Reason / purpose:
read-across source
Key result
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
Potassium iodide did not significantly increase the mutation frequency at any dose tested.
Conclusions:
The read-across analogue test substance potassium iodide did not increase the mutation frequency in the mouse lymphoma L5178Y cells in the absence of metabolic activation up to the concentration of 100 mg/mL. These results can be read across to bismuth oxy-iodide bromide.
Executive summary:

The ability of the read-across analogue test substance potassium iodide to induce gene mutations in mammalian cells in vitro was tested in mouse lymphoma L5178Y cells. The dose which reduced cell viability to 50% in comparison to control values in a preliminary cytotoxicity study was chosen as the highest dose levels. The experiments were performed in the absence of metabolic activation with 4 hours exposure duration. Potassium iodide did not induce the increased mutation frequency at any tested concentration. Positive controls produced satisfactory response. Based on this, potassium iodide was considered to be not mutagenic in mammalian cells in vitro under the conditions of the test. These results can be read across to bismuth oxy-iodide bromide.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

Substance analogue bismuth vanadate was shown to be negative in an in vivo Micronucleus assay . This result is read across to bismuth oxy-iodide bromide. The endpoint conclusion is drawn following a weight-of-evidence approach.

Link to relevant study records
Reference
Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
The rationale to read across the data is attached in section 13.
Reason / purpose:
read-across source
Evaluation criteria:

control value and the range of the historical vehicle control data.
A test substance is considered negative if the following criteria are met:
• The number of cells containing micronuclei in the dose groups is not statistically significant increased above the concurrent vehicle control value and is within the range of the historical vehicle control data.
Key result
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 2000 mg/kg bw
- Clinical signs of toxicity in test animals: piloerection and hunched posture


RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): none
- Ratio of PCE/NCE (for Micronucleus assay): no effect
- Appropriateness of dose levels and route: 2000 mg/kg bw is the highest recommended dose
Conclusions:
Under the experimental conditions chosen here, the test substance BiVO4 (Pigment Yellow 184) has no chromosome-damaging (clastogenic) effect nor does it lead to any impairment of chromosome distribution in the course of mitosis (aneugenic activity) in bone marrow cells in vivo. This result is read across to bismuth oxy-iodide bromide (rationale attached in section 13).
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

An AMES test was performed according to OECD/ EC guidelines and GLP principles. All bacterial strains showed negative responses up to and including 5000µg/plate, i.e. no significant dose-related increase in the number of revertants with or without metabolic activation was seen. No cytotoxicity of the test substance was observed, but the test substance precipitated at 3330 ug/plate and above. The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly. Based on the results of this study it is concluded that bismuth oxy-iodide bromide is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay with or without metabolic activation. No other data on genotoxic properties of the target chemical are available. To address all mode-of-actions, data from analogue bismuth vanadate and data on the individual ions were read acrossed to bismuth oxy-iodide bromide. An in vitro mammalian chromosome aberration test equivalent to OECD Guideline 473 is available for bismuth vanadium tetraoxide. The study was performed using Chinese hamster lung fibroblast (V79) and effects were assessed in absence and presence of metabolic activation. Bismuth vanadium tetraoxide was shown not to induce chromosome aberrations when tested up to cytotoxic concentrations. Furthermore, an in vivo micronucleus test is available performed with bismuth vanadium tetraoxide, based on which it is concluded that BiVO4 has no chromosome-damaging (clastogenic) effect and that it does not lead to any impairment of chromosome distribution in the course of mitosis (aneugenic activity) in bone marrow cells in vivo. Substance analogue sodium bromide was tested in a GLP-compliant study performed according to OECD guideline 482. The test substance did not cause DNA damage in human HeLa-S3 cell line, up to the maximal test concentration of 25,600 µg/mL, both with and without metabolic activation, as measured by silver grain counts (unscheduled DNA synthesis) after an exposure duration of 3 hours. Furthermore, in a GLP-compliant chromosome aberration study performed according to OECD guideline 473, sodium bromide was tested up to the concentration of 5000 µg/mL both in the absence and presence of metabolic activation. Sodium bromide was found not to induce any significant increases either in the proportion of metaphases with chromosome aberrations, or the type of aberrations. Based on these results, sodium bromide showed no evidence of clastogenicity under conditions of the study. Taken together, there are no indications that the bromide ion has genotoxic potential. Iodine is an essential nutrient that humans need for thyroid hormone regulation with a tolerable upper intake level of 600 µg/day for adults including pregnant and lactating women. Therefore, iodine/iodide is not considered to be genotoxic. This was confirmed by two in vitro studies. Potassium iodide (0.1-1.0 mg/ml) did not show mutagenic effects in L5178Y mouse lymphoma cells. Furthermore, potassium iodide also did not induce DNA damage in an in vitro COMET assay. These data further indicate that the iodide ion does not have genotoxic properties.

In conclusion, based on the data available for bismuth oxy-iodide bromide, the separate ions and on an analogue substance, it is concluded that there is sufficient data to conclude that bismuth oxy-iodide bromide does not have genotoxic properties.

Justification for classification or non-classification

Based on the available data, bismuth oxy-iodide bromide is not classified for genotoxic properties according to Regulation (EC) No 1272/2008.