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Administrative data

Description of key information

A local lymph node assay performed with bismuth oxy-iodide bromide according to OECD/EC guidelines and GLP principlesis is available (Klimisch 1 study).

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
(adopted 22 July 2010)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
(updated 06 July 2012)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
dd. 14 September 2015
Type of study:
mouse local lymph node assay (LLNA)
Specific details on test material used for the study:
Dose calculation not adjusted to purity.
Species:
mouse
Strain:
CBA
Remarks:
CaOlaHsd
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS B.V., Inc., Horst, The Netherlands
- Females: nulliparous and non-pregnant
- Age at study initiation: pre-test: 10 - 11 weeks; main study: 8 - 9 weeks
- Weight at study initiation: 17.6-21.4g
- Housing: group-housed in Makrolon Type II (pre-test) or III (main study) cages with wire mesh top
- Diet: 2018C Teklad Global 18% protein rodent diet (certified), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS (set conditions)
- Temperature (°C): 22+/- 2
- Humidity (%): 45-65
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
- IN-LIFE DATES: from 01 June 2016 to 28 June 2016
Vehicle:
propylene glycol
Concentration:
0, 5, 10 and 25%
No. of animals per dose:
5
Details on study design:
PRE-SCREEN TEST:
- Compound solubility: A solubility experiment was performed according to the recommendations given by OECD 429. The highest test item concentration, which could be technically used was a 25% suspension in PG. Grinding of the test item in a mortar was used to formulate the test item.
- Irritation: Two mice were treated by (epidermal) topical application to the dorsal surface of each ear with test item concentrations of 10 and 25% once daily each on three consecutive days.
- Systemic toxicity: Prior to the first application of the test item and before sacrifice the body weight was determined. Clinical signs were recorded at least once daily.
- Ear thickness measurements: Prior to the first application of the test item (day 1), on day 3 and before sacrifice (day 6) the ear thickness was determined using a micrometer. Additionally, for both animals, the ears were punched after sacrifice (day 6) at the apical area using a biopsy punch (Ø 8 mm corresponding to 0.5 cm2) and were immediately pooled per animal and weighed using an analytical balance.
- Erythema scores: Eventual signs of local irritation were documented and a score was used to grade a possible erythema of the ear skin.

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response: A test item is regarded as a sensitiser in the LLNA if the following criteria are fulfilled: First, that exposure to at least one concentration of the test item resulted in an incorporation of 3HTdR at least 3-fold or greater than that recorded in control mice, as indicated by the Stimulation Index. Second, that the data are compatible with a conventional dose response, although allowance must be made (especially at high topical concentrations) for either local toxicity or immunological suppression.

TREATMENT PREPARATION AND ADMINISTRATION: Each test group of mice was treated by (epidermal) topical application to the dorsal surface of each ear. The application volume, 25 μL/ear/day, was spread over the entire dorsal surface of each ear once daily for three consecutive days. A further group of mice (control animals) was treated with an equivalent volume of the relevant vehicle alone (control animals). Five days after the first topical application (day 6) 250 μL of phosphate-buffered saline containing 20.1 μCi of 3H-methyl thymidine (equivalent to 80.2 μCi/mL 3HTdR) were injected into each test and control mouse via the tail vein. Approximately five hours after treatment with 3HTdR all mice were euthanized by using CO2, which was, after harvesting of the lymph nodes, followed by cervical dislocation to ensure death. The draining lymph nodes were rapidly excised and pooled per animal (2 nodes per animal, except for animal 5 for which only one lymph node could be harvested). 3HTdR incorporation was determined with a β-scintillation counter.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
The mean values and standard deviations were calculated in the body weight tables.
Where appropriate, the EC3 value were calculated according to the equation EC3 = (a-c) [(3-d)/(b-d)] + c where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining lymph node cell proliferative activity; (a, b) and (c, d) are respectively the co-ordinates of the two pair of data lying immediately above and below the S.I. value of 3 on the local lymph node assay dose response plot.
All calculations conducted on the DPM values were performed with a validated test script of “R”, a language and environment for statistical computing and graphics. Within the program the Dean-Dixon-Test and Grubb’s Test were used for identification of possible outliers.
Key result
Parameter:
SI
Value:
1.5
Variability:
0.8-2.3
Test group / Remarks:
5%
Key result
Parameter:
SI
Value:
1.8
Variability:
1.1-2.2
Test group / Remarks:
15%
Key result
Parameter:
SI
Value:
1.4
Variability:
0.7-2.2
Test group / Remarks:
25%
Cellular proliferation data / Observations:
CELLULAR PROLIFERATION DATA
Mean DPM per animal were 508.4, 786.4, 898.4 and 699.2 for respectively the vehicle control group, and the groups treated with 5, 10 and 25% bismuth oxy-iodide bromide. Standard deviations were 169.2, 275.5, 294.2 and 320.4 for respectively the vehicle control group, and the groups treated with 5, 10 and 25% bismuth oxy-iodide bromide.

EC3 CALCULATION
The EC3 value could not be calculated, since all S.I.´s are below the threshold value of 3.

CLINICAL OBSERVATIONS:
No deaths occurred during the study period. No symptoms of local skin irritation at the ears of the animals and no signs of systemic toxicity were observed during the study period. From day 1 to 3, redness of the ear skin could not be examined due to the colour of the test item.

BODY WEIGHTS
The body weight gain of the animals was within the range commonly recorded for animals of this strain and age.

No outlier was detected by the Dean-Dixon-Test and Grubb’s Test.

Results positive control 9historical data with a-hexylcinnamalehyde):

Mean DPM per animal were 1042.8, 1201.1, 2908.9 and 8170.5 for respectively the vehicle control group, and the groups treated with 5, 10 and 25% a-hexylcinnamalehyde. Calculated SI values were 1.15, 2.79 and 7.84 for respectively the groups treated with 5, 10 and 25% a-hexylcinnamalehyde. This resulted in an EC3 of 10.6% (w/v).

Results screening test:

At the tested concentrations the animals did not show any signs of local skin irritation or systemic toxicity. It was noted that from day 1 to 4, redness of the ear skin could not be examined due to the colour of the test item.

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of a local lymph node assay performed according to OECD/EC guidelines and GLP principles, it is concluded that bismuth oxy-iodide bromide is not a skin sensitiser under the test conditions of this study.
Executive summary:

A local lymph node assay was performed with bismuth oxy-iodide bromide according to OECD/EC guidelines and GLP principles. The test substance was tested at concentrations of 5, 10 and 25% (in propylene glycol). The highest concentration tested was the highest concentration that could technically be achieved. The animals did not show any signs of systemic toxicity or local skin irritation during the course of the study and no cases of mortality were observed. From day 1 to 3, redness of the ear skin could not be examined due to the colour of the test item. Mean DPM per animal were 508.4, 786.4, 898.4 and 699.2 for respectively the vehicle control group, and the groups treated with 5, 10 and 25% bismuth oxy-iodide bromide. This resulted in Stimulation Indices (S.I.) of 1.5, 1.8, and 1.4 when tested at respectively 5, 10 and 25%. Based on this result, it is concluded that bismuth oxy-iodide bromide is not a skin sensitiser under the test conditions of this study.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

A local lymph node assay was performed with bismuth oxy-iodide bromide according to OECD/EC guidelines and GLP principles. The test substance was tested at concentrations of 5, 10 and 25% (in propylene glycol). The highest concentration tested was the highest concentration that could technically be achieved. The animals did not show any signs of systemic toxicity or local skin irritation during the course of the study and no cases of mortality were observed. From day 1 to 3, redness of the ear skin could not be examined due to the colour of the test item. Mean DPM per animal were 508.4, 786.4, 898.4 and 699.2 for respectively the vehicle control group, and the groups treated with 5, 10 and 25% bismuth oxy-iodide bromide. This resulted in Stimulation Indices (S.I.) of 1.5, 1.8, and 1.4 when tested at respectively 5, 10 and 25%. Based on these results, it is concluded that bismuth oxy-iodide bromide is not a skin sensitiser under the test conditions of this study.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

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