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Description of key information

An in vitro skin irritation test ((EPISKIN Standard model (EPISKIN-SMTM)) was performed with bismuth oxy-iodide bromide according to OECD guideline and GLP principles. Based on this test it is concluded that bismuth oxy-iodide bromide is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.

An in vitro eye irritancy test was performed with bismuth oxy-iodide bromide (20% (w/w) solution) using the Bovine Corneal Opacity and Permeability test (BCOP test) according to OECD/ EC guidelines and GLP principles. It was concluded that bismuth oxy-iodide bromide is not irritant in this test.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 - 23 June 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
Amended by EC No. 761/2009 OJ No. L220, 24 August 2009
Deviations:
no
GLP compliance:
yes
Species:
human
Details on test animals and environmental conditions:
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 36.1 - 37.5
- Humidity (%): 80-95
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amounts applied: approximately 10 mg, 5 μl Milli-Q water to moisten skin.
Duration of treatment / exposure:
15 minutes
Details on study design:
TEST SITE
EPISKIN Standard ModelTM (EPISKIN-SMTM, 0.38 cm2, Lot no.: 10-EKIN-026). This model is a three-dimensional human epidermis model, which consists of adult human derived epidermal keratinocytes which have been seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. The keratinocytes were cultured for 13 days, which results in a highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.
REMOVAL OF TEST SUBSTANCE
Washing: phosphate buffered saline - Time after start of exposure: 15 minutes
POST INCUBATION PERIOD
42 hours
SCORING SYSTEM
Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.
NEGATIVE CONTROL
Phosphate buffered saline (PBS, Invitrogen Corporation, Breda, The Netherlands).
POSITIVE CONTROL
5% (aq) sodium dodecyl sulphate (SDS, Sigma Aldrich, Zwijndrecht, The Netherlands) in PBS.
Irritation / corrosion parameter:
% tissue viability
Value:
100
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
8%
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
The relative mean tissue viability obtained after 15 minutes treatment compared to the negative control tissues was 100%.
The positive control had a mean cell viability after 15 minutes exposure of 8%. The absolute mean OD570 of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 18%, indicating that the test system functioned properly.

Bismuth oxy-iodide bromide was checked for possible direct MTT reduction by adding the test substance to MTT medium. Because no colour change was observed it was concluded that bismuth oxy-iodide bromide did not

interact with MTT.
Interpretation of results:
GHS criteria not met
Conclusions:
An in vitro skin irritation test ((EPISKIN Standard model (EPISKIN-SMTM)) was performed according to OECD guideline and GLP principles. Based on this test it is concluded that bismuth oxy-iodide bromide is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.
Executive summary:

An in vitro skin irritation test ((EPISKIN Standard model (EPISKIN-SMTM)) was performed according to OECD guideline and GLP principles. The relative mean tissue viability obtained after 15 minutes treatment compared to the negative control tissues was 100%. The positive control had a mean cell viability after 15 minutes exposure of 8%. Since the mean relative tissue viability was above 50% after 15 minutes treatment bismuth oxy-iodide bromide

is considered to be non-irritant.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 - 15 July 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
other: OECD guideline 437 “Bovine corneal opacity and permeability (BCOP) test method for identifying ocular corrosives and severe irritants”
Deviations:
no
Qualifier:
according to
Guideline:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 124. Bovine Opacity and Permeability Assay - SOP of Microbiological Associates Ltd., 1999.
Deviations:
no
Qualifier:
according to
Guideline:
other: Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.
Deviations:
no
Principles of method if other than guideline:
pH: 7.5 (50g/ L)
GLP compliance:
yes
Specific details on test material used for the study:
pH: 7.5 (50g/ L)
Vehicle:
physiological saline
Amount / concentration applied:
TEST MATERIAL
- Amount applied (volume or weight with unit): approx. 750 ul of 20% (w/w) test substance solution per cornea

NEGATIVE CONTROL
- Amount applied: 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount applied: 750 µl per cornea
Concentration (if solution): 20% (w/v) Imidazole


Duration of treatment / exposure:
240 minutes
Details on study design:
TEST SITE
- Isolated bovine cornea

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 240 minutes

CONTROLS
Negative control: A negative control, physiological saline (Merck, Darmstadt, Germany) was included to detect non-specific changes in the test system and to provide a baseline for the assay endpoints.
Positive control: 20% (w/w) Imidazole (Merck Schuchardt DHG, Germany) [CAS Number 288-32-4] solution prepared in physiological saline.

SCORING SYSTEM:
- After exposure the cornea is thoroughly rinsed to remove the test substance followed by immediate opacity measurement and permeability evaluation of the cornea. The permeability is evaluated by sodium fluorescein.

- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

TOOL USED TO ASSESS SCORE:
- opacitymeter and microplate reader

Irritation parameter:
in vitro irritation score
Remarks:
240 minutes
Run / experiment:
mean of 3 replica
Value:
ca. 0.7
Negative controls validity:
valid
Remarks:
0
Positive controls validity:
valid
Remarks:
79
Other effects / acceptance of results:
The corneas treated showed opacity values ranging from 0 to 1 and permeability values ranging from 0.020 to 0.057. The corneas were clear after the 240 minutes of treatment. No pH effect of the test substance was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from 0.3 to 1.9 after 240 minutes of treatment. The individual in vitro irritancy scores for the negative controls ranged from -0.9 to 0.0. The individual positive control in vitro irritancy scores ranged from 111 to 139. The corneas treated with the positive control were turbid after the 240 minutes of treatment.
Interpretation of results:
GHS criteria not met
Conclusions:
An in vitro eye irritancy test was performed with bismuth oxy-iodide bromide (20% (w/w) solution) using the Bovine Corneal Opacity and Permeability test (BCOP test) according to OECD/ EC guidelines and GLP principles. It was concluded that bismuth oxy-iodide bromide is not irritant in this test.

Executive summary:

An in vitro eye irritancy test was performed with bismuth oxy-iodide bromide (20% (w/w) solution) using the Bovine Corneal Opacity and Permeability test (BCOP test) according to OECD/EC guidelines and GLP principles.

Reliable positive and negative controls were included. The corneas were clear after the 240 minutes of treatment. The in vitro irritancy scores ranged from 0.3 to 1.9 after 240 minutes of treatment. Based on these data, it is concluded that this test is valid and that bismuth oxy-iodide bromide (tested as a 20% (w/w) solution) is not severe irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

An in vitro skin irritation test ((EPISKIN Standard model (EPISKIN-SMTM)) was performed with bismuth oxy-iodide bromide according to OECD/ EC guidelines and GLP principles. The relative mean tissue viability obtained after 15 minutes treatment compared to the negative control tissues was 100%. The positive control had a mean cell viability after 15 minutes exposure of 8%. Since the mean relative tissue viability was above 50% after 15 minutes treatment bismuth oxy-iodide bromide is considered to be non-irritant.

An in vitro eye irritancy test was performed with bismuth oxy-iodide bromide (20% (w/w) solution) using the Bovine Corneal Opacity and Permeability test (BCOP test) according to OECD/EC guidelines and GLP principles. Reliable positive and negative controls were included. The corneas were clear after the 240 minutes of treatment. The in vitro irritancy scores ranged from 0.3 to 1.9 after 240 minutes of treatment. Based on these data, it is concluded that this test is valid and that bismuth oxy-iodide bromide (tested as a 20% (w/w) solution) is not severe irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Justification for classification or non-classification

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